TY  - CONF
AU  - Stevanović, Danijela
AU  - Paunović, Verica
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Perović, Vladimir
AU  - Ristić, Biljana
AU  - Bošnjak, Mihajlo
AU  - Mandić, Miloš
AU  - Harhaji-Trajković, Ljubica
AU  - Janjetović, Kristina
AU  - Kosić, Milica
AU  - Lalošević, Jovan
AU  - Nikolić, Miloš
AU  - Bonači-Nikolić, Branka
AU  - Trajković, Vladimir
PY  - 2023
UR  - https://indico.bio.bg.ac.rs/event/4/attachments/6/492/Abstract%20Book-CoMBoS2-TMB.pdf
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6286
AB  - Introduction: Since the interaction between autophagy and virus-induced inflammation is complex,
we investigated the interplay between autophagy and inflammation in COVID-19 patients and THP-1
cells expressing SARS-Cov2 proteins NSP5 and ORF3a.
Methods: Autophagy markers in blood from 19 control subjects and 26 COVID-19 patients at hospital
admission and one week later were measured by ELISA, while cytokine levels were examined by flow cytometric bead immunoassay. The level of p62 in cells and its concentration in cell culture supernatants
was measured by immunoblot/ELISA. The mRNA levels of proinflammatory cytokines were measured
by RT-qPCR.
Results: IFN-α, TNF, IL-6, IL-8, IL-17, IL-33, and IFN-γ were elevated in COVID-19 patients at both time
points, whereasIL-10 and IL-1β were elevated at admission and one week later, respectively. Autophagy
markers LC3 and ATG5 were unchanged in COVID-19. The concentration of autophagic cargo receptor
p62 was significantly lower and positively correlated with TNF, IL-10, IL-17, and IL-33 at hospital admission, returning to normal levels after one week. The expression of SARS-CoV-2 proteins NSP5 or ORF3a
in THP-1 cells caused an autophagy-independent decrease/autophagy-inhibition-dependent increase
of intracellular and secreted p62. This was associated with an NSP5-mediated decrease inTNF/IL-10 mRNA
and an ORF3a-mediated increase inTNF/IL-1β/IL-6/IL-10/IL-33 mRNA levels. A genetic knockdown of p62
mimicked the immunosuppressive effect of NSP5, while a p62 increase in autophagy-deficient cells mirrored the immunostimulatory action of ORF3a.
Conclusion: The autophagy receptor p62 is reduced in acute COVID-19, and the balance between autophagy-independent decrease and autophagy blockade-dependent increase of p62 levels could affect
SARS-CoV-induced inflammation.
PB  - Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade
C3  - Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia
T1  - Autophagy receptor P62 regulates SARS-CoV-2-induced inflammation in COVID-19
SP  - 76
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6286
ER  - 

@conference{
author = "Stevanović, Danijela and Paunović, Verica and Vučićević, Ljubica and Misirkić Marjanović, Maja and Perović, Vladimir and Ristić, Biljana and Bošnjak, Mihajlo and Mandić, Miloš and Harhaji-Trajković, Ljubica and Janjetović, Kristina and Kosić, Milica and Lalošević, Jovan and Nikolić, Miloš and Bonači-Nikolić, Branka and Trajković, Vladimir",
year = "2023",
abstract = "Introduction: Since the interaction between autophagy and virus-induced inflammation is complex,
we investigated the interplay between autophagy and inflammation in COVID-19 patients and THP-1
cells expressing SARS-Cov2 proteins NSP5 and ORF3a.
Methods: Autophagy markers in blood from 19 control subjects and 26 COVID-19 patients at hospital
admission and one week later were measured by ELISA, while cytokine levels were examined by flow cytometric bead immunoassay. The level of p62 in cells and its concentration in cell culture supernatants
was measured by immunoblot/ELISA. The mRNA levels of proinflammatory cytokines were measured
by RT-qPCR.
Results: IFN-α, TNF, IL-6, IL-8, IL-17, IL-33, and IFN-γ were elevated in COVID-19 patients at both time
points, whereasIL-10 and IL-1β were elevated at admission and one week later, respectively. Autophagy
markers LC3 and ATG5 were unchanged in COVID-19. The concentration of autophagic cargo receptor
p62 was significantly lower and positively correlated with TNF, IL-10, IL-17, and IL-33 at hospital admission, returning to normal levels after one week. The expression of SARS-CoV-2 proteins NSP5 or ORF3a
in THP-1 cells caused an autophagy-independent decrease/autophagy-inhibition-dependent increase
of intracellular and secreted p62. This was associated with an NSP5-mediated decrease inTNF/IL-10 mRNA
and an ORF3a-mediated increase inTNF/IL-1β/IL-6/IL-10/IL-33 mRNA levels. A genetic knockdown of p62
mimicked the immunosuppressive effect of NSP5, while a p62 increase in autophagy-deficient cells mirrored the immunostimulatory action of ORF3a.
Conclusion: The autophagy receptor p62 is reduced in acute COVID-19, and the balance between autophagy-independent decrease and autophagy blockade-dependent increase of p62 levels could affect
SARS-CoV-induced inflammation.",
publisher = "Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade",
journal = "Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia",
title = "Autophagy receptor P62 regulates SARS-CoV-2-induced inflammation in COVID-19",
pages = "76",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6286"
}

Stevanović, D., Paunović, V., Vučićević, L., Misirkić Marjanović, M., Perović, V., Ristić, B., Bošnjak, M., Mandić, M., Harhaji-Trajković, L., Janjetović, K., Kosić, M., Lalošević, J., Nikolić, M., Bonači-Nikolić, B.,& Trajković, V.. (2023). Autophagy receptor P62 regulates SARS-CoV-2-induced inflammation in COVID-19. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia
Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade., 76.
https://hdl.handle.net/21.15107/rcub_ibiss_6286

Stevanović D, Paunović V, Vučićević L, Misirkić Marjanović M, Perović V, Ristić B, Bošnjak M, Mandić M, Harhaji-Trajković L, Janjetović K, Kosić M, Lalošević J, Nikolić M, Bonači-Nikolić B, Trajković V. Autophagy receptor P62 regulates SARS-CoV-2-induced inflammation in COVID-19. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. 2023;:76.
https://hdl.handle.net/21.15107/rcub_ibiss_6286 .

Stevanović, Danijela, Paunović, Verica, Vučićević, Ljubica, Misirkić Marjanović, Maja, Perović, Vladimir, Ristić, Biljana, Bošnjak, Mihajlo, Mandić, Miloš, Harhaji-Trajković, Ljubica, Janjetović, Kristina, Kosić, Milica, Lalošević, Jovan, Nikolić, Miloš, Bonači-Nikolić, Branka, Trajković, Vladimir, "Autophagy receptor P62 regulates SARS-CoV-2-induced inflammation in COVID-19" in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia (2023):76,
https://hdl.handle.net/21.15107/rcub_ibiss_6286 .

TY  - CONF
AU  - Mandić, Miloš
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Bošnjak, Mihajlo
AU  - Perović, Vladimir
AU  - Janjetović, Kristina
AU  - Paunović, Verica
AU  - Stevanović, Danijela
AU  - Kosić, Milica
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2023
UR  - https://www.sdir.ac.rs/en/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6301
AB  - Background: Reactive oxygen species (ROS) have been implicated in autophagy induction and mitogen activated protein kinases (MAPK) activation which both participate in the differentiation of hematopoietic and leukemic cells. 
We assessed the role of ROS in MAPK activation and autophagy induction in phorbol myristate acetate-(PMA) induced macrophage differentiation of HL-60 leukemia cells. Material and methods: The macrophage markers CD11b, EGR1, 
CSF1R, and IL-8 were assessed by RT-qPCR and flow cytometry. The activation of MAPK was assessed by ERK and JNK immunoblotting, while autophagy was monitored by LC3-II and p62 immunoblotting. Pharmacological inhibition 
was used to determine the role of MAPK and autophagy in HL60 cell differentiation. Intracellular ROS production was determined by flow cytometric analysis of the green fluorescence emitted by non-selective redox-sensitive dye 2',7'-dichlorodihydrofluorescein diacetate. Antioxidant N-acetylcysteine (NAC) was used to determine the role of ROS in MAPK activation, induction of autophagy and HL-60 macrophage differentiation. Results: PMA-triggered differentiation of HL-60 cells into macrophage-like cells was confirmed by elevated expression of macrophage markers 
CD11b, EGR1, CSF1R, and IL-8. The induction of autophagy was demonstrated by the increase of autophagic flux. Pharmacological inhibition of ERK or JNK suppressed PMA-triggered autophagy induction and differentiation of HL-60 cells into macrophage-like cells. PMA increased the intracellular ROS generation and the antioxidant NAC reduced the expression of macrophage markers EGR-1, CSF1R, IL-8 and CD11b in PMA-treated HL-60 cells. NAC also blocked PMA-induced LC3-II and ERK phosphorylation, but only slightly reduced the phosphorylation of JNK and did not affect 
the levels of p62. Conclusion: Our study revealed the partial involvement of ROS in MAPK-dependent autophagy in the differentiation of HL60 cells, indicating ROS/MAPK-mediated autophagy for further investigation in differentiation therapy of AML.
PB  - Belgrade: Serbian Association for Cancer Research
C3  - Proceedings book of The Sixth Congress of The Serbian Association for Cancer Research with international participation: From Collaboration to Innovation in Cancer Research; 2023 Oct 2-4; Belgrade, Serbia
T1  - The role of ROS in MAPK-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells
SP  - 104
EP  - 105
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6301
ER  - 

@conference{
author = "Mandić, Miloš and Misirkić Marjanović, Maja and Vučićević, Ljubica and Bošnjak, Mihajlo and Perović, Vladimir and Janjetović, Kristina and Paunović, Verica and Stevanović, Danijela and Kosić, Milica and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2023",
abstract = "Background: Reactive oxygen species (ROS) have been implicated in autophagy induction and mitogen activated protein kinases (MAPK) activation which both participate in the differentiation of hematopoietic and leukemic cells. 
We assessed the role of ROS in MAPK activation and autophagy induction in phorbol myristate acetate-(PMA) induced macrophage differentiation of HL-60 leukemia cells. Material and methods: The macrophage markers CD11b, EGR1, 
CSF1R, and IL-8 were assessed by RT-qPCR and flow cytometry. The activation of MAPK was assessed by ERK and JNK immunoblotting, while autophagy was monitored by LC3-II and p62 immunoblotting. Pharmacological inhibition 
was used to determine the role of MAPK and autophagy in HL60 cell differentiation. Intracellular ROS production was determined by flow cytometric analysis of the green fluorescence emitted by non-selective redox-sensitive dye 2',7'-dichlorodihydrofluorescein diacetate. Antioxidant N-acetylcysteine (NAC) was used to determine the role of ROS in MAPK activation, induction of autophagy and HL-60 macrophage differentiation. Results: PMA-triggered differentiation of HL-60 cells into macrophage-like cells was confirmed by elevated expression of macrophage markers 
CD11b, EGR1, CSF1R, and IL-8. The induction of autophagy was demonstrated by the increase of autophagic flux. Pharmacological inhibition of ERK or JNK suppressed PMA-triggered autophagy induction and differentiation of HL-60 cells into macrophage-like cells. PMA increased the intracellular ROS generation and the antioxidant NAC reduced the expression of macrophage markers EGR-1, CSF1R, IL-8 and CD11b in PMA-treated HL-60 cells. NAC also blocked PMA-induced LC3-II and ERK phosphorylation, but only slightly reduced the phosphorylation of JNK and did not affect 
the levels of p62. Conclusion: Our study revealed the partial involvement of ROS in MAPK-dependent autophagy in the differentiation of HL60 cells, indicating ROS/MAPK-mediated autophagy for further investigation in differentiation therapy of AML.",
publisher = "Belgrade: Serbian Association for Cancer Research",
journal = "Proceedings book of The Sixth Congress of The Serbian Association for Cancer Research with international participation: From Collaboration to Innovation in Cancer Research; 2023 Oct 2-4; Belgrade, Serbia",
title = "The role of ROS in MAPK-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells",
pages = "104-105",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6301"
}

Mandić, M., Misirkić Marjanović, M., Vučićević, L., Bošnjak, M., Perović, V., Janjetović, K., Paunović, V., Stevanović, D., Kosić, M., Harhaji-Trajković, L.,& Trajković, V.. (2023). The role of ROS in MAPK-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells. in Proceedings book of The Sixth Congress of The Serbian Association for Cancer Research with international participation: From Collaboration to Innovation in Cancer Research; 2023 Oct 2-4; Belgrade, Serbia
Belgrade: Serbian Association for Cancer Research., 104-105.
https://hdl.handle.net/21.15107/rcub_ibiss_6301

Mandić M, Misirkić Marjanović M, Vučićević L, Bošnjak M, Perović V, Janjetović K, Paunović V, Stevanović D, Kosić M, Harhaji-Trajković L, Trajković V. The role of ROS in MAPK-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells. in Proceedings book of The Sixth Congress of The Serbian Association for Cancer Research with international participation: From Collaboration to Innovation in Cancer Research; 2023 Oct 2-4; Belgrade, Serbia. 2023;:104-105.
https://hdl.handle.net/21.15107/rcub_ibiss_6301 .

Mandić, Miloš, Misirkić Marjanović, Maja, Vučićević, Ljubica, Bošnjak, Mihajlo, Perović, Vladimir, Janjetović, Kristina, Paunović, Verica, Stevanović, Danijela, Kosić, Milica, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "The role of ROS in MAPK-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells" in Proceedings book of The Sixth Congress of The Serbian Association for Cancer Research with international participation: From Collaboration to Innovation in Cancer Research; 2023 Oct 2-4; Belgrade, Serbia (2023):104-105,
https://hdl.handle.net/21.15107/rcub_ibiss_6301 .

TY  - CONF
AU  - Mandić, Miloš
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Bošnjak, Mihajlo
AU  - Perović, Vladimir
AU  - Ristić, Biljana
AU  - Ćirić, Darko
AU  - Janjetović, Kristina
AU  - Paunović, Verica
AU  - Stevanović, Danijela
AU  - Kosić, Milica
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2023
UR  - https://indico.bio.bg.ac.rs/event/4/attachments/6/492/Abstract%20Book-CoMBoS2-TMB.pdf
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6285
AB  - Introduction: Autophagy has been shown to participate in the differentiation of hematopoietic and
leukemic cells. We investigated the mechanisms of autophagy action in the differentiation induced by
PKC activator phorbol myristate acetate (PMA) in HL-60 acute myeloid leukemia cells.
Methods: The macrophage markers CD11b, CD13, CD14, CD45, EGR1, CSF1R, and IL-8 were assessed by
flow cytometry and RT-qPCR. Autophagy was monitored by RT-qPCR analysis of autophagy-related (ATG)
gene expression, LC3-II/p62 immunoblotting, beclin-1/Bcl-2 interaction, nuclear translocation of TFEB
and FOXO1/3. The activation of MAP kinases, ERK and JNK was assessed by immunoblotting. Pharmacological inhibition and RNA interference were used to determine the role of MAP kinases and autophagy
in HL60 cell differentiation.
Results: PMA-triggered differentiation of HL-60 cells into macrophage-like cells was confirmed by elevated expression of macrophage markers CD11b, CD13, CD14, CD45, EGR1, CSF1R, and IL-8. The induction of autophagy was demonstrated by accumulation/punctuation of LC3-II, and the increase in
autophagic flux. PMA also increased nuclear translocation of TFEB, FOXO1/3, as well asthe expression of
several ATG genesin HL-60 cells. PMA stimulated the phosphorylation of ERK and JNK via PKC-dependent
mechanism. Pharmacological or genetic inhibition of ERK or JNK suppressed PMA-triggered nuclear
translocation of TFEB and FOXO1/3, ATG expression, dissociation of beclin-1 from Bcl-2, autophagy induction, and differentiation of HL-60 cells into macrophage-like cells.
Conclusion: Our study revealed the involvement of ERK and JNK in TFEB/FOXO-dependent autophagy
and differentiation of HL60 cells, indicating MAP kinase-mediated autophagy as a possible target in differentiation therapy of AML.
PB  - Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade
C3  - Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia
T1  - MAP kinases activate TFEB/FOXO-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells
SP  - 56
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6285
ER  - 

@conference{
author = "Mandić, Miloš and Misirkić Marjanović, Maja and Vučićević, Ljubica and Bošnjak, Mihajlo and Perović, Vladimir and Ristić, Biljana and Ćirić, Darko and Janjetović, Kristina and Paunović, Verica and Stevanović, Danijela and Kosić, Milica and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2023",
abstract = "Introduction: Autophagy has been shown to participate in the differentiation of hematopoietic and
leukemic cells. We investigated the mechanisms of autophagy action in the differentiation induced by
PKC activator phorbol myristate acetate (PMA) in HL-60 acute myeloid leukemia cells.
Methods: The macrophage markers CD11b, CD13, CD14, CD45, EGR1, CSF1R, and IL-8 were assessed by
flow cytometry and RT-qPCR. Autophagy was monitored by RT-qPCR analysis of autophagy-related (ATG)
gene expression, LC3-II/p62 immunoblotting, beclin-1/Bcl-2 interaction, nuclear translocation of TFEB
and FOXO1/3. The activation of MAP kinases, ERK and JNK was assessed by immunoblotting. Pharmacological inhibition and RNA interference were used to determine the role of MAP kinases and autophagy
in HL60 cell differentiation.
Results: PMA-triggered differentiation of HL-60 cells into macrophage-like cells was confirmed by elevated expression of macrophage markers CD11b, CD13, CD14, CD45, EGR1, CSF1R, and IL-8. The induction of autophagy was demonstrated by accumulation/punctuation of LC3-II, and the increase in
autophagic flux. PMA also increased nuclear translocation of TFEB, FOXO1/3, as well asthe expression of
several ATG genesin HL-60 cells. PMA stimulated the phosphorylation of ERK and JNK via PKC-dependent
mechanism. Pharmacological or genetic inhibition of ERK or JNK suppressed PMA-triggered nuclear
translocation of TFEB and FOXO1/3, ATG expression, dissociation of beclin-1 from Bcl-2, autophagy induction, and differentiation of HL-60 cells into macrophage-like cells.
Conclusion: Our study revealed the involvement of ERK and JNK in TFEB/FOXO-dependent autophagy
and differentiation of HL60 cells, indicating MAP kinase-mediated autophagy as a possible target in differentiation therapy of AML.",
publisher = "Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade",
journal = "Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia",
title = "MAP kinases activate TFEB/FOXO-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells",
pages = "56",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6285"
}

Mandić, M., Misirkić Marjanović, M., Vučićević, L., Bošnjak, M., Perović, V., Ristić, B., Ćirić, D., Janjetović, K., Paunović, V., Stevanović, D., Kosić, M., Harhaji-Trajković, L.,& Trajković, V.. (2023). MAP kinases activate TFEB/FOXO-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia
Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade., 56.
https://hdl.handle.net/21.15107/rcub_ibiss_6285

Mandić M, Misirkić Marjanović M, Vučićević L, Bošnjak M, Perović V, Ristić B, Ćirić D, Janjetović K, Paunović V, Stevanović D, Kosić M, Harhaji-Trajković L, Trajković V. MAP kinases activate TFEB/FOXO-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. 2023;:56.
https://hdl.handle.net/21.15107/rcub_ibiss_6285 .

Mandić, Miloš, Misirkić Marjanović, Maja, Vučićević, Ljubica, Bošnjak, Mihajlo, Perović, Vladimir, Ristić, Biljana, Ćirić, Darko, Janjetović, Kristina, Paunović, Verica, Stevanović, Danijela, Kosić, Milica, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "MAP kinases activate TFEB/FOXO-dependent autophagy involved in phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells" in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia (2023):56,
https://hdl.handle.net/21.15107/rcub_ibiss_6285 .

TY  - JOUR
AU  - Paunović, Verica
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Perović, Vladimir
AU  - Ristić, Biljana
AU  - Bošnjak, Mihajlo
AU  - Mandić, Miloš
AU  - Stevanović, Danijela
AU  - Harhaji-Trajković, Ljubica
AU  - Lalošević, Jovan
AU  - Nikolić, Miloš
AU  - Bonači-Nikolić, Branka
AU  - Trajković, Vladimir
PY  - 2023
UR  - https://www.mdpi.com/2073-4409/12/9/1282
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5912
AB  - As autophagy can promote or inhibit inflammation, we examined autophagy-inflammation interplay in COVID-19. Autophagy markers in the blood of 19 control subjects and 26 COVID-19 patients at hospital admission and one week later were measured by ELISA, while cytokine levels were examined by flow cytometric bead immunoassay. The antiviral IFN-α and proinflammatory TNF, IL-6, IL-8, IL-17, IL-33, and IFN-γ were elevated in COVID-19 patients at both time points, while IL-10 and IL-1β were increased at admission and one week later, respectively. Autophagy markers LC3 and ATG5 were unaltered in COVID-19. In contrast, the concentration of autophagic cargo receptor p62 was significantly lower and positively correlated with TNF, IL-10, IL-17, and IL-33 at hospital admission, returning to normal levels after one week. The expression of SARS-CoV-2 proteins NSP5 or ORF3a in THP-1 monocytes caused an autophagy-independent decrease or autophagy-inhibition-dependent increase, respectively, of intracellular/secreted p62, as confirmed by immunoblot/ELISA. This was associated with an NSP5-mediated decrease in TNF/IL-10 mRNA and an ORF3a-mediated increase in TNF/IL-1β/IL-6/IL-10/IL-33 mRNA levels. A genetic knockdown of p62 mimicked the immunosuppressive effect of NSP5, and a p62 increase in autophagy-deficient cells mirrored the immunostimulatory action of ORF3a. In conclusion, the proinflammatory autophagy receptor p62 is reduced inacute COVID-19, and the balance between autophagy-independent decrease and autophagy blockade-dependent increase of p62 levels could affect SARS-CoV-induced inflammation.
PB  - Basel: MDPI
T2  - Cells
T1  - Autophagy Receptor p62 Regulates SARS-CoV-2-Induced Inflammation in COVID-19
IS  - 9
VL  - 12
DO  - 10.3390/cells12091282
SP  - 1282
ER  - 

@article{
author = "Paunović, Verica and Vučićević, Ljubica and Misirkić Marjanović, Maja and Perović, Vladimir and Ristić, Biljana and Bošnjak, Mihajlo and Mandić, Miloš and Stevanović, Danijela and Harhaji-Trajković, Ljubica and Lalošević, Jovan and Nikolić, Miloš and Bonači-Nikolić, Branka and Trajković, Vladimir",
year = "2023",
abstract = "As autophagy can promote or inhibit inflammation, we examined autophagy-inflammation interplay in COVID-19. Autophagy markers in the blood of 19 control subjects and 26 COVID-19 patients at hospital admission and one week later were measured by ELISA, while cytokine levels were examined by flow cytometric bead immunoassay. The antiviral IFN-α and proinflammatory TNF, IL-6, IL-8, IL-17, IL-33, and IFN-γ were elevated in COVID-19 patients at both time points, while IL-10 and IL-1β were increased at admission and one week later, respectively. Autophagy markers LC3 and ATG5 were unaltered in COVID-19. In contrast, the concentration of autophagic cargo receptor p62 was significantly lower and positively correlated with TNF, IL-10, IL-17, and IL-33 at hospital admission, returning to normal levels after one week. The expression of SARS-CoV-2 proteins NSP5 or ORF3a in THP-1 monocytes caused an autophagy-independent decrease or autophagy-inhibition-dependent increase, respectively, of intracellular/secreted p62, as confirmed by immunoblot/ELISA. This was associated with an NSP5-mediated decrease in TNF/IL-10 mRNA and an ORF3a-mediated increase in TNF/IL-1β/IL-6/IL-10/IL-33 mRNA levels. A genetic knockdown of p62 mimicked the immunosuppressive effect of NSP5, and a p62 increase in autophagy-deficient cells mirrored the immunostimulatory action of ORF3a. In conclusion, the proinflammatory autophagy receptor p62 is reduced inacute COVID-19, and the balance between autophagy-independent decrease and autophagy blockade-dependent increase of p62 levels could affect SARS-CoV-induced inflammation.",
publisher = "Basel: MDPI",
journal = "Cells",
title = "Autophagy Receptor p62 Regulates SARS-CoV-2-Induced Inflammation in COVID-19",
number = "9",
volume = "12",
doi = "10.3390/cells12091282",
pages = "1282"
}

Paunović, V., Vučićević, L., Misirkić Marjanović, M., Perović, V., Ristić, B., Bošnjak, M., Mandić, M., Stevanović, D., Harhaji-Trajković, L., Lalošević, J., Nikolić, M., Bonači-Nikolić, B.,& Trajković, V.. (2023). Autophagy Receptor p62 Regulates SARS-CoV-2-Induced Inflammation in COVID-19. in Cells
Basel: MDPI., 12(9), 1282.
https://doi.org/10.3390/cells12091282

Paunović V, Vučićević L, Misirkić Marjanović M, Perović V, Ristić B, Bošnjak M, Mandić M, Stevanović D, Harhaji-Trajković L, Lalošević J, Nikolić M, Bonači-Nikolić B, Trajković V. Autophagy Receptor p62 Regulates SARS-CoV-2-Induced Inflammation in COVID-19. in Cells. 2023;12(9):1282.
doi:10.3390/cells12091282 .

Paunović, Verica, Vučićević, Ljubica, Misirkić Marjanović, Maja, Perović, Vladimir, Ristić, Biljana, Bošnjak, Mihajlo, Mandić, Miloš, Stevanović, Danijela, Harhaji-Trajković, Ljubica, Lalošević, Jovan, Nikolić, Miloš, Bonači-Nikolić, Branka, Trajković, Vladimir, "Autophagy Receptor p62 Regulates SARS-CoV-2-Induced Inflammation in COVID-19" in Cells, 12, no. 9 (2023):1282,
https://doi.org/10.3390/cells12091282 . .

TY  - CONF
AU  - Paunović, Verica
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Stevanović, Danijela
AU  - Ristić, Biljana
AU  - Bošnjak, Mihajlo
AU  - Mandić, Miloš
AU  - Trajković, Vladimir
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6570
AB  - Autophagy is a homeostatic lysosome-dependent catabolic process that eliminates damaged organelles, dysfunctional proteins, and macromolecular aggregates. Autophagy plays an important role in host response to viral infection as it enables degradation of viruses in autophagolysosomes and regulates innate and adaptive immunity. However, some viruses, including SARS-CoV-2, have evolved a variety of mechanisms to avoid autophagic degradation and use it for their own benefit. The aim of this study is to investigate the impact of the individual SARS-CoV-2 proteins (M, E, N, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12, NSP14, and NSP15) on autophagy in human lung epithelial cells by analyzing the expression of autophagy-related proteins, LC3-II, p62, and beclin1. The immunoblot analysis revealed that intracellular expression of non-structural proteins NSP4, NSP6, and NSP8 increased the levels of autophagy markers LC3-II and beclin-1, while the structural N protein and non-structural proteins NSP5, NSP10, and NSP15, reduced the degradation of autophagy-selective target p62. These data indicate that some SARS-CoV-2 proteins induce autophagic response, while others block its completion, thus providing grounds for further investigation of the complex interaction between the virus and the autophagic pathway.
AB  - Аутофагија је лизозомски посредован хомеостатски катаболички процес током којег долази до елиминисања оштећених органела, дисфункционалних протеина и макромолекуларних комплекса. Аутофагија игра важну улогу у одговору домаћина на вирусну инфекцију јер омогућава деградацију вируса у аутофаголизозомима и регулише урођени и стечени имунитет. Међутим, неки вируси, укључујући и SARS-CoV-2, су развили различите механизме како би избегли деградацију која се дешава током процеса аутофагије и подредили је у своју корист. Ова студија има за циљ да испита утицај појединачних SARS-CoV-2 протеина (M, E, N, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12, NSP14 и NSP15) на процес аутофагије који се одвија у ћелијама респираторног епитела код људи анализом експресије протеина повезаних са аутофагијом, LC3-II, p62, и беклин 1. Имуноблот анализа је открила да је унутарћелијска експресија неструктурних протеина NSP4, NSP6 и NSP8 повећала нивое маркера аутофагије LC3-II и беклин-1, док су структурни N протеин и неструктурни протеини NSP5, NSP10 и NSP15 довели до смањења деградације рецептора аутофагије p62. Ови подаци указују на то да неки SARS-CoV-2 протеини индукују аутофагни одговор, док други блокирају завршетак процеса аутофагије, чиме се ствара основа за даље истраживање комплексне интеракције између вируса и процеса аутофагије.
PB  - Beograd: Srpska akademija nauka i umetnosti
C3  - Proceedings: COVID-19 Pandemic: Messages, New Information and Dilemmas; 2021 Jun 4; Belgrade, Serbia
T1  - Modulation of autophagy by SARS-CoV-2 proteins
T1  - Модулација аутофагије SARS-CoV-2 протеинима
SP  - 205
EP  - 212
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6570
ER  - 

@conference{
author = "Paunović, Verica and Misirkić Marjanović, Maja and Vučićević, Ljubica and Stevanović, Danijela and Ristić, Biljana and Bošnjak, Mihajlo and Mandić, Miloš and Trajković, Vladimir",
year = "2022",
abstract = "Autophagy is a homeostatic lysosome-dependent catabolic process that eliminates damaged organelles, dysfunctional proteins, and macromolecular aggregates. Autophagy plays an important role in host response to viral infection as it enables degradation of viruses in autophagolysosomes and regulates innate and adaptive immunity. However, some viruses, including SARS-CoV-2, have evolved a variety of mechanisms to avoid autophagic degradation and use it for their own benefit. The aim of this study is to investigate the impact of the individual SARS-CoV-2 proteins (M, E, N, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12, NSP14, and NSP15) on autophagy in human lung epithelial cells by analyzing the expression of autophagy-related proteins, LC3-II, p62, and beclin1. The immunoblot analysis revealed that intracellular expression of non-structural proteins NSP4, NSP6, and NSP8 increased the levels of autophagy markers LC3-II and beclin-1, while the structural N protein and non-structural proteins NSP5, NSP10, and NSP15, reduced the degradation of autophagy-selective target p62. These data indicate that some SARS-CoV-2 proteins induce autophagic response, while others block its completion, thus providing grounds for further investigation of the complex interaction between the virus and the autophagic pathway., Аутофагија је лизозомски посредован хомеостатски катаболички процес током којег долази до елиминисања оштећених органела, дисфункционалних протеина и макромолекуларних комплекса. Аутофагија игра важну улогу у одговору домаћина на вирусну инфекцију јер омогућава деградацију вируса у аутофаголизозомима и регулише урођени и стечени имунитет. Међутим, неки вируси, укључујући и SARS-CoV-2, су развили различите механизме како би избегли деградацију која се дешава током процеса аутофагије и подредили је у своју корист. Ова студија има за циљ да испита утицај појединачних SARS-CoV-2 протеина (M, E, N, NSP4, NSP5, NSP6, NSP7, NSP8, NSP10, NSP12, NSP14 и NSP15) на процес аутофагије који се одвија у ћелијама респираторног епитела код људи анализом експресије протеина повезаних са аутофагијом, LC3-II, p62, и беклин 1. Имуноблот анализа је открила да је унутарћелијска експресија неструктурних протеина NSP4, NSP6 и NSP8 повећала нивое маркера аутофагије LC3-II и беклин-1, док су структурни N протеин и неструктурни протеини NSP5, NSP10 и NSP15 довели до смањења деградације рецептора аутофагије p62. Ови подаци указују на то да неки SARS-CoV-2 протеини индукују аутофагни одговор, док други блокирају завршетак процеса аутофагије, чиме се ствара основа за даље истраживање комплексне интеракције између вируса и процеса аутофагије.",
publisher = "Beograd: Srpska akademija nauka i umetnosti",
journal = "Proceedings: COVID-19 Pandemic: Messages, New Information and Dilemmas; 2021 Jun 4; Belgrade, Serbia",
title = "Modulation of autophagy by SARS-CoV-2 proteins, Модулација аутофагије SARS-CoV-2 протеинима",
pages = "205-212",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6570"
}

Paunović, V., Misirkić Marjanović, M., Vučićević, L., Stevanović, D., Ristić, B., Bošnjak, M., Mandić, M.,& Trajković, V.. (2022). Modulation of autophagy by SARS-CoV-2 proteins. in Proceedings: COVID-19 Pandemic: Messages, New Information and Dilemmas; 2021 Jun 4; Belgrade, Serbia
Beograd: Srpska akademija nauka i umetnosti., 205-212.
https://hdl.handle.net/21.15107/rcub_ibiss_6570

Paunović V, Misirkić Marjanović M, Vučićević L, Stevanović D, Ristić B, Bošnjak M, Mandić M, Trajković V. Modulation of autophagy by SARS-CoV-2 proteins. in Proceedings: COVID-19 Pandemic: Messages, New Information and Dilemmas; 2021 Jun 4; Belgrade, Serbia. 2022;:205-212.
https://hdl.handle.net/21.15107/rcub_ibiss_6570 .

Paunović, Verica, Misirkić Marjanović, Maja, Vučićević, Ljubica, Stevanović, Danijela, Ristić, Biljana, Bošnjak, Mihajlo, Mandić, Miloš, Trajković, Vladimir, "Modulation of autophagy by SARS-CoV-2 proteins" in Proceedings: COVID-19 Pandemic: Messages, New Information and Dilemmas; 2021 Jun 4; Belgrade, Serbia (2022):205-212,
https://hdl.handle.net/21.15107/rcub_ibiss_6570 .

TY  - JOUR
AU  - Mandić, Miloš
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Jovanović, Maja
AU  - Bošnjak, Mihajlo
AU  - Perović, Vladimir
AU  - Ristić, Biljana
AU  - Ćirić, Darko
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2022
UR  - https://linkinghub.elsevier.com/retrieve/pii/S0024320522001813
UR  - http://www.ncbi.nlm.nih.gov/pubmed/35304128
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4947
AB  - We investigated the mechanisms and the role of autophagy in the differentiation of HL-60 human acute myeloid leukemia cells induced by protein kinase C (PKC) activator phorbol myristate acetate (PMA). PMA-triggered differentiation of HL-60 cells into macrophage-like cells was confirmed by cell-cycle arrest accompanied by elevated expression of macrophage markers CD11b, CD13, CD14, CD45, EGR1, CSF1R, and IL-8. The induction of autophagy was demonstrated by the increase in intracellular acidification, accumulation/punctuation of autophagosome marker LC3-II, and the increase in autophagic flux. PMA also increased nuclear translocation of autophagy transcription factors TFEB, FOXO1, and FOXO3, as well as the expression of several autophagy-related (ATG) genes in HL-60 cells. PMA failed to activate autophagy inducer AMP-activated protein kinase (AMPK) and inhibit autophagy suppressor mechanistic target of rapamycin complex 1 (mTORC1). On the other hand, it readily stimulated the phosphorylation of mitogen-activated protein (MAP) kinases extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) via a protein kinase C-dependent mechanism. Pharmacological or genetic inhibition of ERK or JNK suppressed PMA-triggered nuclear translocation of TFEB and FOXO1/3, ATG expression, dissociation of pro-autophagic beclin-1 from its inhibitor BCL2, autophagy induction, and differentiation of HL-60 cells into macrophage-like cells. Pharmacological or genetic inhibition of autophagy also blocked PMA-induced macrophage differentiation of HL-60 cells. Therefore, MAP kinases ERK and JNK control PMA-induced macrophage differentiation of HL-60 leukemia cells through AMPK/mTORC1-independent, TFEB/FOXO-mediated transcriptional and beclin-1-dependent post-translational activation of autophagy.
PB  - Elsevier Inc.
T2  - Life Sciences
T1  - MAP kinase-dependent autophagy controls phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells.
VL  - 297
DO  - 10.1016/j.lfs.2022.120481
SP  - 120481
ER  - 

@article{
author = "Mandić, Miloš and Misirkić Marjanović, Maja and Vučićević, Ljubica and Jovanović, Maja and Bošnjak, Mihajlo and Perović, Vladimir and Ristić, Biljana and Ćirić, Darko and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2022",
abstract = "We investigated the mechanisms and the role of autophagy in the differentiation of HL-60 human acute myeloid leukemia cells induced by protein kinase C (PKC) activator phorbol myristate acetate (PMA). PMA-triggered differentiation of HL-60 cells into macrophage-like cells was confirmed by cell-cycle arrest accompanied by elevated expression of macrophage markers CD11b, CD13, CD14, CD45, EGR1, CSF1R, and IL-8. The induction of autophagy was demonstrated by the increase in intracellular acidification, accumulation/punctuation of autophagosome marker LC3-II, and the increase in autophagic flux. PMA also increased nuclear translocation of autophagy transcription factors TFEB, FOXO1, and FOXO3, as well as the expression of several autophagy-related (ATG) genes in HL-60 cells. PMA failed to activate autophagy inducer AMP-activated protein kinase (AMPK) and inhibit autophagy suppressor mechanistic target of rapamycin complex 1 (mTORC1). On the other hand, it readily stimulated the phosphorylation of mitogen-activated protein (MAP) kinases extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) via a protein kinase C-dependent mechanism. Pharmacological or genetic inhibition of ERK or JNK suppressed PMA-triggered nuclear translocation of TFEB and FOXO1/3, ATG expression, dissociation of pro-autophagic beclin-1 from its inhibitor BCL2, autophagy induction, and differentiation of HL-60 cells into macrophage-like cells. Pharmacological or genetic inhibition of autophagy also blocked PMA-induced macrophage differentiation of HL-60 cells. Therefore, MAP kinases ERK and JNK control PMA-induced macrophage differentiation of HL-60 leukemia cells through AMPK/mTORC1-independent, TFEB/FOXO-mediated transcriptional and beclin-1-dependent post-translational activation of autophagy.",
publisher = "Elsevier Inc.",
journal = "Life Sciences",
title = "MAP kinase-dependent autophagy controls phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells.",
volume = "297",
doi = "10.1016/j.lfs.2022.120481",
pages = "120481"
}

Mandić, M., Misirkić Marjanović, M., Vučićević, L., Jovanović, M., Bošnjak, M., Perović, V., Ristić, B., Ćirić, D., Harhaji-Trajković, L.,& Trajković, V.. (2022). MAP kinase-dependent autophagy controls phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells.. in Life Sciences
Elsevier Inc.., 297, 120481.
https://doi.org/10.1016/j.lfs.2022.120481

Mandić M, Misirkić Marjanović M, Vučićević L, Jovanović M, Bošnjak M, Perović V, Ristić B, Ćirić D, Harhaji-Trajković L, Trajković V. MAP kinase-dependent autophagy controls phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells.. in Life Sciences. 2022;297:120481.
doi:10.1016/j.lfs.2022.120481 .

Mandić, Miloš, Misirkić Marjanović, Maja, Vučićević, Ljubica, Jovanović, Maja, Bošnjak, Mihajlo, Perović, Vladimir, Ristić, Biljana, Ćirić, Darko, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "MAP kinase-dependent autophagy controls phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells." in Life Sciences, 297 (2022):120481,
https://doi.org/10.1016/j.lfs.2022.120481 . .

TY  - CONF
AU  - Janjetović, Kristina
AU  - Stamenković, Marina
AU  - Tovilović-Kovačević, Gordana
AU  - Zogović, Nevena
AU  - Despotović, Ana
AU  - Stevanović, Danijela
AU  - Mandić, Miloš
AU  - Kosić, Milica
AU  - Paunović, Verica
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Trajković, Vladimir
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5737
AB  - I pored stalnog napretka lečenja kancera, ova bolest ostaje druga po smrtnosti u svetu. Kako bi se skratio vremenski i finansijski zahtevan proces razvoja novih hemoterapeutika poslednjih desetak godina intezivno se radi na ispitivanju antikancerskog potencijala lekova koji se već koriste u terapiji drugih bolesti. U ovom radu smo proučavali potencijalni antikancerski efekat inhibitora protonske pumpe pantoprazola (PPZ), terapeutika koji se standardno koristi u lečenju kiselinskih gastrointestinalnih poremećaja. Citotoksični efekat PPZ je ispitivan u kulturama humanog U251 glioblastoma, humanog H460 nesitnoćelijskog karcinoma pluća i mišjeg B16 melanoma. Pokazano je da PPZ aktiviranoj apoptozi u svim ispitivanim ćelijskim linijama prethodi povećana produkcija reaktivnih vrsta kiseonika, depolarizacija mitohondrija i aktivacija kaspaza. U prisustvu PPZ detektovano je povećanje LC3 II proteina ukazujući na aktivaciju autofagije. Detaljnijim ispitivanjem mehanizma koji je u osnovi toksičnog efekta PPZ, utvrđeno je da PPZ aktivira AKT/AMPK signalni put u ispitivanim ćelijskim linijama i stimuliše AMPK zavisnu citoprotektivnu autofagiju u U251 i B16 ćelijskim linijama. Sa druge strane, autofagija aktivirana u ćelijama karcinoma pluća je citotoksična. Sumirano, PPZ ispoljava značajan antikancerski potencijal prema U251, H460 i B16 ćelijama izazivajući apoptozu, pri čemu uloga autofagije u smrti ćelija može biti citoprotektivna ili citotoksična i zavisi od tipa ćelija. Dodatna farmakološka modulacija autofagije mogla bi poboljšati antikancerski potencijal pantoprazola.
AB  - И поред сталног напретка лечења канцера, ова болест остаје друга по смртности у
свету. Како би се скратио временски и финансијски захтеван процес развоја нових
хемотерапеутика последњих десетак година интезивно се ради на испитивању
антиканцерског потенцијала лекова који се већ користе у терапији других болести.
У овом раду смо проучавали потенцијални антиканцерски ефекат инхибитора
протонске пумпе пантопразола (ППЗ), терапеутика који се стандардно користи у
лечењу киселинских гастроинтестиналних поремећаја. Цитотоксични ефекат ППЗ
је испитиван у културама хуманог U251 глиобластома, хуманог H460
неситноћелијског карцинома плућа и мишјег B16 меланома. Показано је да ППЗ
активираној апоптози у свим испитиваним ћелијским линијама претходи повећана
продукција реактивних врста кисеоника, деполаризација митохондрија и
активација каспаза. У присуству ППЗ детектовано је повећање LC3 II протеина
указујући на активацију аутофагије. Детаљнијим испитивањем механизма који је у
основи токсичног ефекта ППЗ, утврђено је да ППЗ активира AKT/AMPK сигнални
пут у испитиваним ћелијским линијама и стимулише AMPK зависну
цитопротективну аутофагију у U251 и B16 ћелијским линијама. Са друге стране,
аутофагија активирана у ћелијама карцинома плућа је цитотоксична. Сумирано,
ППЗ испољава значајан антиканцерски потенцијал према U251, H460 и B16
ћелијама изазивајући апоптозу, при чему улога аутофагије у смрти ћелија може
бити цитопротективна или цитотоксична и зависи од типа ћелија. Додатна
фармаколошка модулација аутофагије могла би побољшати антиканцерски
потенцијал пантопразола.
PB  - Belgrade: Serbian Biological Society
C3  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - Antikancerski potencijal inhibitora protonske pumpe pantoprazola
T1  - Антиканцерски потенцијал инхибитора протонске пумпе пантопразола
SP  - 285
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5737
ER  - 

@conference{
author = "Janjetović, Kristina and Stamenković, Marina and Tovilović-Kovačević, Gordana and Zogović, Nevena and Despotović, Ana and Stevanović, Danijela and Mandić, Miloš and Kosić, Milica and Paunović, Verica and Vučićević, Ljubica and Misirkić Marjanović, Maja and Trajković, Vladimir",
year = "2022",
abstract = "I pored stalnog napretka lečenja kancera, ova bolest ostaje druga po smrtnosti u svetu. Kako bi se skratio vremenski i finansijski zahtevan proces razvoja novih hemoterapeutika poslednjih desetak godina intezivno se radi na ispitivanju antikancerskog potencijala lekova koji se već koriste u terapiji drugih bolesti. U ovom radu smo proučavali potencijalni antikancerski efekat inhibitora protonske pumpe pantoprazola (PPZ), terapeutika koji se standardno koristi u lečenju kiselinskih gastrointestinalnih poremećaja. Citotoksični efekat PPZ je ispitivan u kulturama humanog U251 glioblastoma, humanog H460 nesitnoćelijskog karcinoma pluća i mišjeg B16 melanoma. Pokazano je da PPZ aktiviranoj apoptozi u svim ispitivanim ćelijskim linijama prethodi povećana produkcija reaktivnih vrsta kiseonika, depolarizacija mitohondrija i aktivacija kaspaza. U prisustvu PPZ detektovano je povećanje LC3 II proteina ukazujući na aktivaciju autofagije. Detaljnijim ispitivanjem mehanizma koji je u osnovi toksičnog efekta PPZ, utvrđeno je da PPZ aktivira AKT/AMPK signalni put u ispitivanim ćelijskim linijama i stimuliše AMPK zavisnu citoprotektivnu autofagiju u U251 i B16 ćelijskim linijama. Sa druge strane, autofagija aktivirana u ćelijama karcinoma pluća je citotoksična. Sumirano, PPZ ispoljava značajan antikancerski potencijal prema U251, H460 i B16 ćelijama izazivajući apoptozu, pri čemu uloga autofagije u smrti ćelija može biti citoprotektivna ili citotoksična i zavisi od tipa ćelija. Dodatna farmakološka modulacija autofagije mogla bi poboljšati antikancerski potencijal pantoprazola., И поред сталног напретка лечења канцера, ова болест остаје друга по смртности у
свету. Како би се скратио временски и финансијски захтеван процес развоја нових
хемотерапеутика последњих десетак година интезивно се ради на испитивању
антиканцерског потенцијала лекова који се већ користе у терапији других болести.
У овом раду смо проучавали потенцијални антиканцерски ефекат инхибитора
протонске пумпе пантопразола (ППЗ), терапеутика који се стандардно користи у
лечењу киселинских гастроинтестиналних поремећаја. Цитотоксични ефекат ППЗ
је испитиван у културама хуманог U251 глиобластома, хуманог H460
неситноћелијског карцинома плућа и мишјег B16 меланома. Показано је да ППЗ
активираној апоптози у свим испитиваним ћелијским линијама претходи повећана
продукција реактивних врста кисеоника, деполаризација митохондрија и
активација каспаза. У присуству ППЗ детектовано је повећање LC3 II протеина
указујући на активацију аутофагије. Детаљнијим испитивањем механизма који је у
основи токсичног ефекта ППЗ, утврђено је да ППЗ активира AKT/AMPK сигнални
пут у испитиваним ћелијским линијама и стимулише AMPK зависну
цитопротективну аутофагију у U251 и B16 ћелијским линијама. Са друге стране,
аутофагија активирана у ћелијама карцинома плућа је цитотоксична. Сумирано,
ППЗ испољава значајан антиканцерски потенцијал према U251, H460 и B16
ћелијама изазивајући апоптозу, при чему улога аутофагије у смрти ћелија може
бити цитопротективна или цитотоксична и зависи од типа ћелија. Додатна
фармаколошка модулација аутофагије могла би побољшати антиканцерски
потенцијал пантопразола.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia",
title = "Antikancerski potencijal inhibitora protonske pumpe pantoprazola, Антиканцерски потенцијал инхибитора протонске пумпе пантопразола",
pages = "285",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5737"
}

Janjetović, K., Stamenković, M., Tovilović-Kovačević, G., Zogović, N., Despotović, A., Stevanović, D., Mandić, M., Kosić, M., Paunović, V., Vučićević, L., Misirkić Marjanović, M.,& Trajković, V.. (2022). Antikancerski potencijal inhibitora protonske pumpe pantoprazola. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 285.
https://hdl.handle.net/21.15107/rcub_ibiss_5737

Janjetović K, Stamenković M, Tovilović-Kovačević G, Zogović N, Despotović A, Stevanović D, Mandić M, Kosić M, Paunović V, Vučićević L, Misirkić Marjanović M, Trajković V. Antikancerski potencijal inhibitora protonske pumpe pantoprazola. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:285.
https://hdl.handle.net/21.15107/rcub_ibiss_5737 .

Janjetović, Kristina, Stamenković, Marina, Tovilović-Kovačević, Gordana, Zogović, Nevena, Despotović, Ana, Stevanović, Danijela, Mandić, Miloš, Kosić, Milica, Paunović, Verica, Vučićević, Ljubica, Misirkić Marjanović, Maja, Trajković, Vladimir, "Antikancerski potencijal inhibitora protonske pumpe pantoprazola" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):285,
https://hdl.handle.net/21.15107/rcub_ibiss_5737 .

TY  - CONF
AU  - Kosić, Milica
AU  - Paunović, Verica
AU  - Ristić, Biljana
AU  - Mirčić, Aleksandar
AU  - Bošnjak, Mihajlo
AU  - Stevanović, Danijela
AU  - Mandić, Miloš
AU  - Stamenković, Marina
AU  - Janjetović, Kristina
AU  - Vučićević, Ljubica
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
PY  - 2021
UR  - https://www.sfrre2021belgrade.rs/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4727
AB  - There is no effective therapy for melanoma, a malignant tumor of melanocytes with an
increasing incidence. High energy demands of melanoma cells are predominantly satisfied by
aerobic glycolysis. When glycolysis is suppressed, these metabolically plastic cells switch to
oxidative phosphorylation. The aim of this study was to investigate the antimelanoma effects of
simultaneous inhibition of glycolysis by 2-deoxy-D-glucose (2DG) and oxidative phosphorylation
by rotenone (ROT). 2DG synergized with ROT in inducing death of B16 melanoma, but not
primary mesenchymal cells. Combined treatment stimulated caspase activation, but not PARP
cleavage and DNA fragmentation. Disintegration of plasma membrane and inability of caspase
inhibitors and necrostatin to suppress toxicity of 2DG/ROT implied that combined treatment
induced necrosis, rather than apoptosis and necroptosis. 2DG/ROT stimulated ATP depletion,
mitochondrial superoxide production, and mitochondrial swelling, but not depolarization
of mitochondria. 2DG/ROT-induced toxicity was suppressed by antioxidant α-tocopherol,
but not mitochondrial depolarization inhibitor cyclosporine. Combined treatment induced
the translocation of hexokinase II, a suppressor of voltage-dependent anion channel (VDAC)
opening, and cytochrome c from mitochondria in the cytoplasm, while VDAC opening inhibitor
DIDS suppressed 2DG/ROT toxicity. Our results suggest that 2DG/ROT treatment stimulates
mitochondrial swelling, release of hexokinase II and subsequent opening of VDAC in the outer
mitochondrial membrane. These events allow cytochrome c to exit and activate caspases, which
are unable to stimulate PARP and consequent DNA fragmentation in the energy-depleted state.
On the other hand, superoxide synthesized in mitochondria upon 2DG/ROT treatment also exits
through VDAC and triggers energy-independent necrosis. Simultaneous inhibition of glycolysis
and oxidative phosphorylation appears to be promising strategy for further development of
novel anticancer therapeutics.
PB  - Elsevier Inc.
C3  - Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia
T1  - Synergistic anticancer effect of glycolysis inhibition and oxidative phosphorylation suppression
DO  - 10.1016/j.freeradbiomed.2021.08.205
SP  - 203
ER  - 

@conference{
author = "Kosić, Milica and Paunović, Verica and Ristić, Biljana and Mirčić, Aleksandar and Bošnjak, Mihajlo and Stevanović, Danijela and Mandić, Miloš and Stamenković, Marina and Janjetović, Kristina and Vučićević, Ljubica and Trajković, Vladimir and Harhaji-Trajković, Ljubica",
year = "2021",
abstract = "There is no effective therapy for melanoma, a malignant tumor of melanocytes with an
increasing incidence. High energy demands of melanoma cells are predominantly satisfied by
aerobic glycolysis. When glycolysis is suppressed, these metabolically plastic cells switch to
oxidative phosphorylation. The aim of this study was to investigate the antimelanoma effects of
simultaneous inhibition of glycolysis by 2-deoxy-D-glucose (2DG) and oxidative phosphorylation
by rotenone (ROT). 2DG synergized with ROT in inducing death of B16 melanoma, but not
primary mesenchymal cells. Combined treatment stimulated caspase activation, but not PARP
cleavage and DNA fragmentation. Disintegration of plasma membrane and inability of caspase
inhibitors and necrostatin to suppress toxicity of 2DG/ROT implied that combined treatment
induced necrosis, rather than apoptosis and necroptosis. 2DG/ROT stimulated ATP depletion,
mitochondrial superoxide production, and mitochondrial swelling, but not depolarization
of mitochondria. 2DG/ROT-induced toxicity was suppressed by antioxidant α-tocopherol,
but not mitochondrial depolarization inhibitor cyclosporine. Combined treatment induced
the translocation of hexokinase II, a suppressor of voltage-dependent anion channel (VDAC)
opening, and cytochrome c from mitochondria in the cytoplasm, while VDAC opening inhibitor
DIDS suppressed 2DG/ROT toxicity. Our results suggest that 2DG/ROT treatment stimulates
mitochondrial swelling, release of hexokinase II and subsequent opening of VDAC in the outer
mitochondrial membrane. These events allow cytochrome c to exit and activate caspases, which
are unable to stimulate PARP and consequent DNA fragmentation in the energy-depleted state.
On the other hand, superoxide synthesized in mitochondria upon 2DG/ROT treatment also exits
through VDAC and triggers energy-independent necrosis. Simultaneous inhibition of glycolysis
and oxidative phosphorylation appears to be promising strategy for further development of
novel anticancer therapeutics.",
publisher = "Elsevier Inc.",
journal = "Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia",
title = "Synergistic anticancer effect of glycolysis inhibition and oxidative phosphorylation suppression",
doi = "10.1016/j.freeradbiomed.2021.08.205",
pages = "203"
}

Kosić, M., Paunović, V., Ristić, B., Mirčić, A., Bošnjak, M., Stevanović, D., Mandić, M., Stamenković, M., Janjetović, K., Vučićević, L., Trajković, V.,& Harhaji-Trajković, L.. (2021). Synergistic anticancer effect of glycolysis inhibition and oxidative phosphorylation suppression. in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia
Elsevier Inc.., 203.
https://doi.org/10.1016/j.freeradbiomed.2021.08.205

Kosić M, Paunović V, Ristić B, Mirčić A, Bošnjak M, Stevanović D, Mandić M, Stamenković M, Janjetović K, Vučićević L, Trajković V, Harhaji-Trajković L. Synergistic anticancer effect of glycolysis inhibition and oxidative phosphorylation suppression. in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia. 2021;:203.
doi:10.1016/j.freeradbiomed.2021.08.205 .

Kosić, Milica, Paunović, Verica, Ristić, Biljana, Mirčić, Aleksandar, Bošnjak, Mihajlo, Stevanović, Danijela, Mandić, Miloš, Stamenković, Marina, Janjetović, Kristina, Vučićević, Ljubica, Trajković, Vladimir, Harhaji-Trajković, Ljubica, "Synergistic anticancer effect of glycolysis inhibition and oxidative phosphorylation suppression" in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia (2021):203,
https://doi.org/10.1016/j.freeradbiomed.2021.08.205 . .

TY  - CONF
AU  - Ristić, Biljana
AU  - Krunić, Matija
AU  - Bošnjak, Mihajlo
AU  - Paunović, Verica
AU  - Zogović, Nevena
AU  - Tovilović-Kovačević, Gordana
AU  - Mirčić, Aleksandar
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Kosić, Milica
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
PY  - 2021
UR  - https://www.sfrre2021belgrade.rs/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4726
AB  - We here investigated the ability of graphene quantum dots (GQD), graphene nanoparticles with antioxidative capacity, to protect SH-SY5Y human neuroblastoma cells from oxidative/nitrosative stress induced by iron-nitrosyl complex sodium nitroprusside (SNP). Although GQD diminished the levels of nitric oxide (NO) in both cell free condition and SNPexposed cells, NO scavengers (PTIO and uric acid), displayed only slight protection from SNP, suggesting that NO scavenging was not the main protective mechanism of GQD. Moreover, GQD significantly protected SH-SY5Y cells from neurotoxicity of light exhausted SNP, incapable of producing NO, implying the existence of protective mechanism independent of NO-scavenging. GQD lowered the increase in the concentration of hydroxyl radical (•OH) and superoxide anion (O2•−) caused by SNP both in the cell-free condition and inside cells, as well as ensuing oxidative stress and lipid peroxidation. Nonspecific antioxidants (glutathione, NAC), •OH scavenger (DMSO), and iron chelators (DTPA, BPDSA), but not superoxide dismutase, mimicked the cytoprotective activity of GQD, suggesting that GQD protect cells by neutralizing •OH generated in the presence of iron released from SNP. GQD were readily internalized by SH-SY5Y cells, while extensive washing of cells pre-incubated with GQD only partly reduced their protective activity, suggesting that GQD exerted neuroprotective effect both intra- and extracellularly. By demonstrating that GQD protect neuroblastoma cells from SNP-induced neurotoxicity by both extracellular •OH/NO scavenging and some unknown intracellular mechanism, our results suggest that GQD could be valuable candidate for treatment of neurodegenerative and neuroinflammatory disorders associated with oxidative/nitrosative stress.
PB  - Elsevier Inc.
C3  - Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia
T1  - Graphene quantum dots protect SH-SY5Y cells from SNP-induced neurotoxicity by ROS/RNS scavenging
DO  - 10.1016/j.freeradbiomed.2021.08.167
SP  - 165
ER  - 

@conference{
author = "Ristić, Biljana and Krunić, Matija and Bošnjak, Mihajlo and Paunović, Verica and Zogović, Nevena and Tovilović-Kovačević, Gordana and Mirčić, Aleksandar and Misirkić Marjanović, Maja and Vučićević, Ljubica and Kosić, Milica and Trajković, Vladimir and Harhaji-Trajković, Ljubica",
year = "2021",
abstract = "We here investigated the ability of graphene quantum dots (GQD), graphene nanoparticles with antioxidative capacity, to protect SH-SY5Y human neuroblastoma cells from oxidative/nitrosative stress induced by iron-nitrosyl complex sodium nitroprusside (SNP). Although GQD diminished the levels of nitric oxide (NO) in both cell free condition and SNPexposed cells, NO scavengers (PTIO and uric acid), displayed only slight protection from SNP, suggesting that NO scavenging was not the main protective mechanism of GQD. Moreover, GQD significantly protected SH-SY5Y cells from neurotoxicity of light exhausted SNP, incapable of producing NO, implying the existence of protective mechanism independent of NO-scavenging. GQD lowered the increase in the concentration of hydroxyl radical (•OH) and superoxide anion (O2•−) caused by SNP both in the cell-free condition and inside cells, as well as ensuing oxidative stress and lipid peroxidation. Nonspecific antioxidants (glutathione, NAC), •OH scavenger (DMSO), and iron chelators (DTPA, BPDSA), but not superoxide dismutase, mimicked the cytoprotective activity of GQD, suggesting that GQD protect cells by neutralizing •OH generated in the presence of iron released from SNP. GQD were readily internalized by SH-SY5Y cells, while extensive washing of cells pre-incubated with GQD only partly reduced their protective activity, suggesting that GQD exerted neuroprotective effect both intra- and extracellularly. By demonstrating that GQD protect neuroblastoma cells from SNP-induced neurotoxicity by both extracellular •OH/NO scavenging and some unknown intracellular mechanism, our results suggest that GQD could be valuable candidate for treatment of neurodegenerative and neuroinflammatory disorders associated with oxidative/nitrosative stress.",
publisher = "Elsevier Inc.",
journal = "Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia",
title = "Graphene quantum dots protect SH-SY5Y cells from SNP-induced neurotoxicity by ROS/RNS scavenging",
doi = "10.1016/j.freeradbiomed.2021.08.167",
pages = "165"
}

Ristić, B., Krunić, M., Bošnjak, M., Paunović, V., Zogović, N., Tovilović-Kovačević, G., Mirčić, A., Misirkić Marjanović, M., Vučićević, L., Kosić, M., Trajković, V.,& Harhaji-Trajković, L.. (2021). Graphene quantum dots protect SH-SY5Y cells from SNP-induced neurotoxicity by ROS/RNS scavenging. in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia
Elsevier Inc.., 165.
https://doi.org/10.1016/j.freeradbiomed.2021.08.167

Ristić B, Krunić M, Bošnjak M, Paunović V, Zogović N, Tovilović-Kovačević G, Mirčić A, Misirkić Marjanović M, Vučićević L, Kosić M, Trajković V, Harhaji-Trajković L. Graphene quantum dots protect SH-SY5Y cells from SNP-induced neurotoxicity by ROS/RNS scavenging. in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia. 2021;:165.
doi:10.1016/j.freeradbiomed.2021.08.167 .

Ristić, Biljana, Krunić, Matija, Bošnjak, Mihajlo, Paunović, Verica, Zogović, Nevena, Tovilović-Kovačević, Gordana, Mirčić, Aleksandar, Misirkić Marjanović, Maja, Vučićević, Ljubica, Kosić, Milica, Trajković, Vladimir, Harhaji-Trajković, Ljubica, "Graphene quantum dots protect SH-SY5Y cells from SNP-induced neurotoxicity by ROS/RNS scavenging" in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia (2021):165,
https://doi.org/10.1016/j.freeradbiomed.2021.08.167 . .

TY  - CONF
AU  - Stevanović, Danijela
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Paunović, Verica
AU  - Kosić, Milica
AU  - Mandić, Miloš
AU  - Ristić, Biljana
AU  - Bošnjak, Mihajlo
AU  - Janjetović, Kristina
AU  - Zogović, Nevena
AU  - Tovilović-Kovačević, Gordana
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2021
UR  - https://www.sfrre2021belgrade.rs/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4725
AB  - 6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenylpyridinium (MPP+) are the most common neurotoxins used to induce experimental model of Parkinson’s disease both in vivo and in vitro. Neurotoxic action of 6-OHDA and MPP+
 is mediated by oxidative stress, mitochondrial damage and induction of apoptotic cell death. Natural disaccharide trehalose exhibits antioxidative properties and stimulates removal of damaged proteins, and thus exhibits powerful
neuroprotective effect in certain brain injury models. We investigated the effects of trehalose in 6-OHDA and MPP+
 - induced oxidative stress and neurotoxicity in human neuroblastoma SH-SY5Y cells. The effects of trehalose on the cell viability and death were assessed by MTT, crystal violet, lactate dehydrogenase assay and AnnexinV-FITC/propidium iodide staining. The production of reactive oxygen species was analyzed by flow cytometry using redox-sensitive dyes dihydrorhodamine 123 (DHR) and MitoSOX Red. Further, activation of stress-related MAP kinases, p38 and JNK were investigated by immunoblot analysis. Our study demonstrated that trehalose pretreatment significantly improved cell viability and reduced neurotoxic effect of 6-OHDA, while slightly decreased cell viability and increased neurotoxic effect of MPP+. Trehalose decreased the number of 6-OHDA-induced apoptotic cells (shown by the reduced % of Annexin V+ and AnnexinV+ PI+ cells) whereas it increased apoptosis in MPP+ treated cells. Flow
cytometric analysis of DHR and MitoSOX stained cells demonstrated that trehalose pretreatment significantly reduced 6-OHDA-triggered ROS and superoxide anion radical generation. However, in MPP+-treated neurons trehalose augmented oxidative stress and production of superoxide anion. Immunoblot analysis showed that trehalose significantly decreased p38 and JNK activation only in 6-OHDA treated cells. These results indicate that trehalose has different effects on oxidative stress induced by two different neurotoxins, 6-OHDA and MPP+, and suggests further
exploration of the mechanism of its antioxidative action.
PB  - Elsevier Inc.
C3  - Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia
T1  - The opposite effects of trehalose on 6-hydroxydopamine and 1-methyl-4- phenylpyridinium induced oxidative stress in human neuroblastoma SH-SY5Y cells
DO  - 10.1016/j.freeradbiomed.2021.08.097
SP  - 94
ER  - 

@conference{
author = "Stevanović, Danijela and Vučićević, Ljubica and Misirkić Marjanović, Maja and Paunović, Verica and Kosić, Milica and Mandić, Miloš and Ristić, Biljana and Bošnjak, Mihajlo and Janjetović, Kristina and Zogović, Nevena and Tovilović-Kovačević, Gordana and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2021",
abstract = "6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenylpyridinium (MPP+) are the most common neurotoxins used to induce experimental model of Parkinson’s disease both in vivo and in vitro. Neurotoxic action of 6-OHDA and MPP+
 is mediated by oxidative stress, mitochondrial damage and induction of apoptotic cell death. Natural disaccharide trehalose exhibits antioxidative properties and stimulates removal of damaged proteins, and thus exhibits powerful
neuroprotective effect in certain brain injury models. We investigated the effects of trehalose in 6-OHDA and MPP+
 - induced oxidative stress and neurotoxicity in human neuroblastoma SH-SY5Y cells. The effects of trehalose on the cell viability and death were assessed by MTT, crystal violet, lactate dehydrogenase assay and AnnexinV-FITC/propidium iodide staining. The production of reactive oxygen species was analyzed by flow cytometry using redox-sensitive dyes dihydrorhodamine 123 (DHR) and MitoSOX Red. Further, activation of stress-related MAP kinases, p38 and JNK were investigated by immunoblot analysis. Our study demonstrated that trehalose pretreatment significantly improved cell viability and reduced neurotoxic effect of 6-OHDA, while slightly decreased cell viability and increased neurotoxic effect of MPP+. Trehalose decreased the number of 6-OHDA-induced apoptotic cells (shown by the reduced % of Annexin V+ and AnnexinV+ PI+ cells) whereas it increased apoptosis in MPP+ treated cells. Flow
cytometric analysis of DHR and MitoSOX stained cells demonstrated that trehalose pretreatment significantly reduced 6-OHDA-triggered ROS and superoxide anion radical generation. However, in MPP+-treated neurons trehalose augmented oxidative stress and production of superoxide anion. Immunoblot analysis showed that trehalose significantly decreased p38 and JNK activation only in 6-OHDA treated cells. These results indicate that trehalose has different effects on oxidative stress induced by two different neurotoxins, 6-OHDA and MPP+, and suggests further
exploration of the mechanism of its antioxidative action.",
publisher = "Elsevier Inc.",
journal = "Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia",
title = "The opposite effects of trehalose on 6-hydroxydopamine and 1-methyl-4- phenylpyridinium induced oxidative stress in human neuroblastoma SH-SY5Y cells",
doi = "10.1016/j.freeradbiomed.2021.08.097",
pages = "94"
}

Stevanović, D., Vučićević, L., Misirkić Marjanović, M., Paunović, V., Kosić, M., Mandić, M., Ristić, B., Bošnjak, M., Janjetović, K., Zogović, N., Tovilović-Kovačević, G., Harhaji-Trajković, L.,& Trajković, V.. (2021). The opposite effects of trehalose on 6-hydroxydopamine and 1-methyl-4- phenylpyridinium induced oxidative stress in human neuroblastoma SH-SY5Y cells. in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia
Elsevier Inc.., 94.
https://doi.org/10.1016/j.freeradbiomed.2021.08.097

Stevanović D, Vučićević L, Misirkić Marjanović M, Paunović V, Kosić M, Mandić M, Ristić B, Bošnjak M, Janjetović K, Zogović N, Tovilović-Kovačević G, Harhaji-Trajković L, Trajković V. The opposite effects of trehalose on 6-hydroxydopamine and 1-methyl-4- phenylpyridinium induced oxidative stress in human neuroblastoma SH-SY5Y cells. in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia. 2021;:94.
doi:10.1016/j.freeradbiomed.2021.08.097 .

Stevanović, Danijela, Vučićević, Ljubica, Misirkić Marjanović, Maja, Paunović, Verica, Kosić, Milica, Mandić, Miloš, Ristić, Biljana, Bošnjak, Mihajlo, Janjetović, Kristina, Zogović, Nevena, Tovilović-Kovačević, Gordana, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "The opposite effects of trehalose on 6-hydroxydopamine and 1-methyl-4- phenylpyridinium induced oxidative stress in human neuroblastoma SH-SY5Y cells" in Free Radical Research Europe (SFRR-E) Annual Meeting Abstracts “Redox biology in the 21st century: a new scientific discipline” 15-18 June 2021, Belgrade, Serbia (2021):94,
https://doi.org/10.1016/j.freeradbiomed.2021.08.097 . .

TY  - JOUR
AU  - Dinić, Jelena
AU  - Harhaji-Trajković, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4174
AB  - In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.
PB  - Informa UK Limited
T2  - Autophagy
T1  - Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)
DO  - 10.1080/15548627.2020.1797280
ER  - 

@article{
author = "Dinić, Jelena and Harhaji-Trajković, Ljubica and Misirkić Marjanović, Maja and Vučićević, Ljubica",
year = "2021",
abstract = "In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.",
publisher = "Informa UK Limited",
journal = "Autophagy",
title = "Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)",
doi = "10.1080/15548627.2020.1797280"
}

Dinić, J., Harhaji-Trajković, L., Misirkić Marjanović, M.,& Vučićević, L.. (2021). Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition). in Autophagy
Informa UK Limited..
https://doi.org/10.1080/15548627.2020.1797280

Dinić J, Harhaji-Trajković L, Misirkić Marjanović M, Vučićević L. Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition). in Autophagy. 2021;.
doi:10.1080/15548627.2020.1797280 .

Dinić, Jelena, Harhaji-Trajković, Ljubica, Misirkić Marjanović, Maja, Vučićević, Ljubica, "Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)" in Autophagy (2021),
https://doi.org/10.1080/15548627.2020.1797280 . .

TY  - JOUR
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Despotović, Ana
AU  - Stamenković, Marina
AU  - Janjetović, Kristina
PY  - 2021
UR  - http://www.ajcr.us/files/ajcr0136757.pdf
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4679
AB  - Metformin has been known to treat type 2 diabetes for decades and is widely prescribed antidiabetic drug.
Recently, its anticancer potential has also been discovered. Moreover, metformin has low cost thus it has attained profound research interest. Comprehensing the complexity of the molecular regulatory networks in cancer provides a mode for advancement of research in cancer development and treatment. Metformin targets many pathways that play an important role in cancer cell survival outcome. Here, we described anticancer activity of metformin on the AMPK dependent/independent mechanisms regulating metabolism, oncogene/tumor suppressor signaling pathways together with the issue of clinical studies. We also provided brief overwiev about recently described metformin’s role in cancer immunity. Insight in these complex molecular networks, will simplify application of metformin in clinical trials and contribute to improvement of anti-cancer therapy.
PB  - Madison, USA : e-Century Publishing Corporation
T2  - American Journal of Cancer Research
T1  - Dual anticancer role of metformin: an old drug regulating AMPK dependent/independent pathways in metabolic, oncogenic/tumorsuppresing and immunity context
IS  - 11
VL  - 11
SP  - 5625
EP  - 5643
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4679
ER  - 

@article{
author = "Misirkić Marjanović, Maja and Vučićević, Ljubica and Despotović, Ana and Stamenković, Marina and Janjetović, Kristina",
year = "2021",
abstract = "Metformin has been known to treat type 2 diabetes for decades and is widely prescribed antidiabetic drug.
Recently, its anticancer potential has also been discovered. Moreover, metformin has low cost thus it has attained profound research interest. Comprehensing the complexity of the molecular regulatory networks in cancer provides a mode for advancement of research in cancer development and treatment. Metformin targets many pathways that play an important role in cancer cell survival outcome. Here, we described anticancer activity of metformin on the AMPK dependent/independent mechanisms regulating metabolism, oncogene/tumor suppressor signaling pathways together with the issue of clinical studies. We also provided brief overwiev about recently described metformin’s role in cancer immunity. Insight in these complex molecular networks, will simplify application of metformin in clinical trials and contribute to improvement of anti-cancer therapy.",
publisher = "Madison, USA : e-Century Publishing Corporation",
journal = "American Journal of Cancer Research",
title = "Dual anticancer role of metformin: an old drug regulating AMPK dependent/independent pathways in metabolic, oncogenic/tumorsuppresing and immunity context",
number = "11",
volume = "11",
pages = "5625-5643",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4679"
}

Misirkić Marjanović, M., Vučićević, L., Despotović, A., Stamenković, M.,& Janjetović, K.. (2021). Dual anticancer role of metformin: an old drug regulating AMPK dependent/independent pathways in metabolic, oncogenic/tumorsuppresing and immunity context. in American Journal of Cancer Research
Madison, USA : e-Century Publishing Corporation., 11(11), 5625-5643.
https://hdl.handle.net/21.15107/rcub_ibiss_4679

Misirkić Marjanović M, Vučićević L, Despotović A, Stamenković M, Janjetović K. Dual anticancer role of metformin: an old drug regulating AMPK dependent/independent pathways in metabolic, oncogenic/tumorsuppresing and immunity context. in American Journal of Cancer Research. 2021;11(11):5625-5643.
https://hdl.handle.net/21.15107/rcub_ibiss_4679 .

Misirkić Marjanović, Maja, Vučićević, Ljubica, Despotović, Ana, Stamenković, Marina, Janjetović, Kristina, "Dual anticancer role of metformin: an old drug regulating AMPK dependent/independent pathways in metabolic, oncogenic/tumorsuppresing and immunity context" in American Journal of Cancer Research, 11, no. 11 (2021):5625-5643,
https://hdl.handle.net/21.15107/rcub_ibiss_4679 .

TY  - JOUR
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Ćirić, Darko
AU  - Martinović, Tamara
AU  - Jovanović, Maja
AU  - Isaković, Aleksandra
AU  - Marković, Ivanka
AU  - Šaponjić, Jasna
AU  - Foretz, Marc
AU  - Rabanal-Ruiz, Yoana
AU  - Korolchuk, Viktor I.
AU  - Trajković, Vladimir
PY  - 2020
UR  - http://link.springer.com/10.1007/s00018-019-03356-2
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3528
AB  - We investigated the role of autophagy, a controlled lysosomal degradation of cellular macromolecules and organelles, in glutamate excitotoxicity during nutrient deprivation in vitro. The incubation in low-glucose serum/amino acid-free cell culture medium synergized with glutamate in increasing AMP/ATP ratio and causing excitotoxic necrosis in SH-SY5Y human neuroblastoma cells. Glutamate suppressed starvation-triggered autophagy, as confirmed by diminished intracellular acidification, lower LC3 punctuation and LC3-I conversion to autophagosome-associated LC3-II, reduced expression of proautophagic beclin-1 and ATG5, increase of the selective autophagic target NBR1, and decreased number of autophagic vesicles. Similar results were observed in PC12 rat pheochromocytoma cells. Both glutamate-mediated excitotoxicity and autophagy inhibition in starved SH-SY5Y cells were reverted by NMDA antagonist memantine and mimicked by NMDA agonists D-aspartate and ibotenate. Glutamate reduced starvation-triggered phosphorylation of the energy sensor AMP-activated protein kinase (AMPK) without affecting the activity of mammalian target of rapamycin complex 1, a major negative regulator of autophagy. This was associated with reduced mRNA levels of autophagy transcriptional activators (FOXO3, ATF4) and molecules involved in autophagy initiation (ULK1, ATG13, FIP200), autophagosome nucleation/elongation (ATG14, beclin-1, ATG5), and autophagic cargo delivery to autophagosomes (SQSTM1). Glutamate-mediated transcriptional repression of autophagy was alleviated by overexpression of constitutively active AMPK. Genetic or pharmacological AMPK activation by AMPK overexpression or metformin, as well as genetic or pharmacological autophagy induction by TFEB overexpression or lithium chloride, reduced the sensitivity of nutrient-deprived SH-SY5Y cells to glutamate excitotoxicity. These data indicate that transcriptional inhibition of AMPK-dependent cytoprotective autophagy is involved in glutamate-mediated excitotoxicity during nutrient deprivation in vitro.
T2  - Cellular and Molecular Life Sciences
T1  - Transcriptional block of AMPK-induced autophagy promotes glutamate excitotoxicity in nutrient-deprived SH-SY5Y neuroblastoma cells.
VL  - 77
DO  - 10.1007/s00018-019-03356-2
SP  - 3383
EP  - 3399
ER  - 

@article{
author = "Vučićević, Ljubica and Misirkić Marjanović, Maja and Ćirić, Darko and Martinović, Tamara and Jovanović, Maja and Isaković, Aleksandra and Marković, Ivanka and Šaponjić, Jasna and Foretz, Marc and Rabanal-Ruiz, Yoana and Korolchuk, Viktor I. and Trajković, Vladimir",
year = "2020",
abstract = "We investigated the role of autophagy, a controlled lysosomal degradation of cellular macromolecules and organelles, in glutamate excitotoxicity during nutrient deprivation in vitro. The incubation in low-glucose serum/amino acid-free cell culture medium synergized with glutamate in increasing AMP/ATP ratio and causing excitotoxic necrosis in SH-SY5Y human neuroblastoma cells. Glutamate suppressed starvation-triggered autophagy, as confirmed by diminished intracellular acidification, lower LC3 punctuation and LC3-I conversion to autophagosome-associated LC3-II, reduced expression of proautophagic beclin-1 and ATG5, increase of the selective autophagic target NBR1, and decreased number of autophagic vesicles. Similar results were observed in PC12 rat pheochromocytoma cells. Both glutamate-mediated excitotoxicity and autophagy inhibition in starved SH-SY5Y cells were reverted by NMDA antagonist memantine and mimicked by NMDA agonists D-aspartate and ibotenate. Glutamate reduced starvation-triggered phosphorylation of the energy sensor AMP-activated protein kinase (AMPK) without affecting the activity of mammalian target of rapamycin complex 1, a major negative regulator of autophagy. This was associated with reduced mRNA levels of autophagy transcriptional activators (FOXO3, ATF4) and molecules involved in autophagy initiation (ULK1, ATG13, FIP200), autophagosome nucleation/elongation (ATG14, beclin-1, ATG5), and autophagic cargo delivery to autophagosomes (SQSTM1). Glutamate-mediated transcriptional repression of autophagy was alleviated by overexpression of constitutively active AMPK. Genetic or pharmacological AMPK activation by AMPK overexpression or metformin, as well as genetic or pharmacological autophagy induction by TFEB overexpression or lithium chloride, reduced the sensitivity of nutrient-deprived SH-SY5Y cells to glutamate excitotoxicity. These data indicate that transcriptional inhibition of AMPK-dependent cytoprotective autophagy is involved in glutamate-mediated excitotoxicity during nutrient deprivation in vitro.",
journal = "Cellular and Molecular Life Sciences",
title = "Transcriptional block of AMPK-induced autophagy promotes glutamate excitotoxicity in nutrient-deprived SH-SY5Y neuroblastoma cells.",
volume = "77",
doi = "10.1007/s00018-019-03356-2",
pages = "3383-3399"
}

Vučićević, L., Misirkić Marjanović, M., Ćirić, D., Martinović, T., Jovanović, M., Isaković, A., Marković, I., Šaponjić, J., Foretz, M., Rabanal-Ruiz, Y., Korolchuk, V. I.,& Trajković, V.. (2020). Transcriptional block of AMPK-induced autophagy promotes glutamate excitotoxicity in nutrient-deprived SH-SY5Y neuroblastoma cells.. in Cellular and Molecular Life Sciences, 77, 3383-3399.
https://doi.org/10.1007/s00018-019-03356-2

Vučićević L, Misirkić Marjanović M, Ćirić D, Martinović T, Jovanović M, Isaković A, Marković I, Šaponjić J, Foretz M, Rabanal-Ruiz Y, Korolchuk VI, Trajković V. Transcriptional block of AMPK-induced autophagy promotes glutamate excitotoxicity in nutrient-deprived SH-SY5Y neuroblastoma cells.. in Cellular and Molecular Life Sciences. 2020;77:3383-3399.
doi:10.1007/s00018-019-03356-2 .

Vučićević, Ljubica, Misirkić Marjanović, Maja, Ćirić, Darko, Martinović, Tamara, Jovanović, Maja, Isaković, Aleksandra, Marković, Ivanka, Šaponjić, Jasna, Foretz, Marc, Rabanal-Ruiz, Yoana, Korolchuk, Viktor I., Trajković, Vladimir, "Transcriptional block of AMPK-induced autophagy promotes glutamate excitotoxicity in nutrient-deprived SH-SY5Y neuroblastoma cells." in Cellular and Molecular Life Sciences, 77 (2020):3383-3399,
https://doi.org/10.1007/s00018-019-03356-2 . .

TY  - CONF
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Ćirić, Darko
AU  - Martinović, Tamara
AU  - Jovanović, Maja
AU  - Isaković, Aleksandra
AU  - Marković, Ivanka
AU  - Trajković, Vladimir
PY  - 2019
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6661
AB  - We investigated the role of autophagy in glutamate excitotoxicity during nutrient deprivation in vitro. Lack of serum, amino acids, and glucose markedly increased the sensitivity of SH-SY5Y human neuroblastoma cell line to glutamate-induced excitotoxic necrosis. Glutamate suppressed starvation-triggered autophagic response, as confirmed by diminished intracellular acidification, lower LC3 punctuation and conversion of LC3I to autophagosome associated LC3II, reduced levels of autophagy activators beclin-1 and ATG5, increased levels of the selective autophagic target NBR1, and reduced appearance of autophagic vesicles observed by transmission electron microscopy. Glutamate reduced starvation-triggered phosphorylation of the intracellular energy sensor AMP-activated protein kinase (AMPK), without affecting the activity of mammalian target of rapamycin complex1 as a major negative regulator of autophagy. Similar results were shown on PC12 cells, which are often exploited as a model for excitotoxicity. We also detected reduced mRNA expression of autophagy transcription factors FOXO3 and ATF4, as well as molecules involved in autophagy initiation (ULK1, ATG13, FIP200), autophagosome nucleation/elongation (ATG14, beclin 1, ATG5, ATG12), and the autophagy cargo delivery to autophagosmes (SQSTM1/p62). Genetic or pharmacological AMPK activation by AMPK overexpression or metformin, reduced the sensitivity of nutrient-deprived SH-SY5Y cells to glutamate dependent autophagy is involved in glutamate-mediated excitotoxicity during nutrient deprivation in vitro.
PB  - Nordic Autophagy Society
C3  - 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Nederlands
T1  - Glutamate-mediated autophagy inhibition intensifies excitotoxic death of nutrient-deprived SH-SY5Y neuroblastoma cells
SP  - 34
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6661
ER  - 

@conference{
author = "Misirkić Marjanović, Maja and Vučićević, Ljubica and Ćirić, Darko and Martinović, Tamara and Jovanović, Maja and Isaković, Aleksandra and Marković, Ivanka and Trajković, Vladimir",
year = "2019",
abstract = "We investigated the role of autophagy in glutamate excitotoxicity during nutrient deprivation in vitro. Lack of serum, amino acids, and glucose markedly increased the sensitivity of SH-SY5Y human neuroblastoma cell line to glutamate-induced excitotoxic necrosis. Glutamate suppressed starvation-triggered autophagic response, as confirmed by diminished intracellular acidification, lower LC3 punctuation and conversion of LC3I to autophagosome associated LC3II, reduced levels of autophagy activators beclin-1 and ATG5, increased levels of the selective autophagic target NBR1, and reduced appearance of autophagic vesicles observed by transmission electron microscopy. Glutamate reduced starvation-triggered phosphorylation of the intracellular energy sensor AMP-activated protein kinase (AMPK), without affecting the activity of mammalian target of rapamycin complex1 as a major negative regulator of autophagy. Similar results were shown on PC12 cells, which are often exploited as a model for excitotoxicity. We also detected reduced mRNA expression of autophagy transcription factors FOXO3 and ATF4, as well as molecules involved in autophagy initiation (ULK1, ATG13, FIP200), autophagosome nucleation/elongation (ATG14, beclin 1, ATG5, ATG12), and the autophagy cargo delivery to autophagosmes (SQSTM1/p62). Genetic or pharmacological AMPK activation by AMPK overexpression or metformin, reduced the sensitivity of nutrient-deprived SH-SY5Y cells to glutamate dependent autophagy is involved in glutamate-mediated excitotoxicity during nutrient deprivation in vitro.",
publisher = "Nordic Autophagy Society",
journal = "3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Nederlands",
title = "Glutamate-mediated autophagy inhibition intensifies excitotoxic death of nutrient-deprived SH-SY5Y neuroblastoma cells",
pages = "34",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6661"
}

Misirkić Marjanović, M., Vučićević, L., Ćirić, D., Martinović, T., Jovanović, M., Isaković, A., Marković, I.,& Trajković, V.. (2019). Glutamate-mediated autophagy inhibition intensifies excitotoxic death of nutrient-deprived SH-SY5Y neuroblastoma cells. in 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Nederlands
Nordic Autophagy Society., 34.
https://hdl.handle.net/21.15107/rcub_ibiss_6661

Misirkić Marjanović M, Vučićević L, Ćirić D, Martinović T, Jovanović M, Isaković A, Marković I, Trajković V. Glutamate-mediated autophagy inhibition intensifies excitotoxic death of nutrient-deprived SH-SY5Y neuroblastoma cells. in 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Nederlands. 2019;:34.
https://hdl.handle.net/21.15107/rcub_ibiss_6661 .

Misirkić Marjanović, Maja, Vučićević, Ljubica, Ćirić, Darko, Martinović, Tamara, Jovanović, Maja, Isaković, Aleksandra, Marković, Ivanka, Trajković, Vladimir, "Glutamate-mediated autophagy inhibition intensifies excitotoxic death of nutrient-deprived SH-SY5Y neuroblastoma cells" in 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Nederlands (2019):34,
https://hdl.handle.net/21.15107/rcub_ibiss_6661 .

TY  - CONF
AU  - Martinović, Tamara
AU  - Ćirić, Darko
AU  - Pantić, Igor
AU  - Lalić, Katarina
AU  - Rasulić, Iva
AU  - Despotović, Sanja
AU  - Lalić, Ivana
AU  - Đuričić, Danica
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Trajković, Vladimir
AU  - Bumbaširević, Vladimir
AU  - Kravić-Stevović, Tamara
PY  - 2019
UR  - https://klinbiolabmed.rs/wp-content/uploads/2022/05/Knjiga-sazetka-Kongresa.pdf
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6662
AB  - Uvod: Osobine tip 2 dijabetes melitusa (T2DM) su insulinska rezistencija, poremećena sekrecija insulina i hiperglikemija. Kao početna terapija T2DM koristi se metformin. Novija istraživanja su pokazala da u T2DM dolazi do morfoloških promena u izgledu nukleusa u vidu nepravilnosti oblika i binukelacije nukleusa.
Cilj: Cilj istraživanja je da se analiziraju ultrastrukturne karakteristike nukleusa limfocita periferne krvi kod pacijenata sa T2DM pomoću kompjuterizovane analize slike, fraktalne i teksturalne analize, kao i utvrđivanje efekta metformina na karakteristike nukleusa.
Metodologija: Mononuklearne ćelije izolovane iz periferne krvi novootkrivenih T2DM bolesnika, bolesnika lečenih metforminom i zdravih ispitanika analizirane su na transmisionom elektronskom mikroskopu (TEM). Primenom ImageJ programa analizirani su oblik i procenat heterohromatina, kao i fraktalna i teksturalna analiza nukleusa limfocita.
Rezultati: Limfociti zdravih osoba su imali okrugle, predominantno heterohromatične nukleuse i malu količinu citoplazme sa retko prisutnim organelama, dok su limfociti T2DM bolesnika imali euhromatične nukleu uz povećanje strukturnih praznina kod T2DM bolesnika. Nivo glukoze našte i HbA1c koreliraju sa fraktalnom dimenzijom i sa parametrima oblika nukleusa. Postoji korelacija između nivoa triglicerida u krvi i fraktalne dimenzije nukleusa.
Zaključak: Nukleusi limfocita bolesnika sa T2DM su nepravilnog oblika i sa većom količinom euhromatina, a promena njihovog izgleda je u vezi sa nivoom glikemije.
PB  - Beograd: Srpsko lekarsko društvo, Sekcija kliničke biohemije
C3  - Knjiga sažetaka: 1. kongres kliničkih biohemičara i specijalista laboratorijske medicine Srbije sa međunarodnim učešćem;  2019 Nov 27-29; Belgrade, Serbia
T1  - Uticaj glikemije i lipidnog statusa na morfološke karakteristike nukleusa kod pacijenata sa tip 2 dijabetes melitusom
SP  - 58
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6662
ER  - 

@conference{
author = "Martinović, Tamara and Ćirić, Darko and Pantić, Igor and Lalić, Katarina and Rasulić, Iva and Despotović, Sanja and Lalić, Ivana and Đuričić, Danica and Vučićević, Ljubica and Misirkić Marjanović, Maja and Trajković, Vladimir and Bumbaširević, Vladimir and Kravić-Stevović, Tamara",
year = "2019",
abstract = "Uvod: Osobine tip 2 dijabetes melitusa (T2DM) su insulinska rezistencija, poremećena sekrecija insulina i hiperglikemija. Kao početna terapija T2DM koristi se metformin. Novija istraživanja su pokazala da u T2DM dolazi do morfoloških promena u izgledu nukleusa u vidu nepravilnosti oblika i binukelacije nukleusa.
Cilj: Cilj istraživanja je da se analiziraju ultrastrukturne karakteristike nukleusa limfocita periferne krvi kod pacijenata sa T2DM pomoću kompjuterizovane analize slike, fraktalne i teksturalne analize, kao i utvrđivanje efekta metformina na karakteristike nukleusa.
Metodologija: Mononuklearne ćelije izolovane iz periferne krvi novootkrivenih T2DM bolesnika, bolesnika lečenih metforminom i zdravih ispitanika analizirane su na transmisionom elektronskom mikroskopu (TEM). Primenom ImageJ programa analizirani su oblik i procenat heterohromatina, kao i fraktalna i teksturalna analiza nukleusa limfocita.
Rezultati: Limfociti zdravih osoba su imali okrugle, predominantno heterohromatične nukleuse i malu količinu citoplazme sa retko prisutnim organelama, dok su limfociti T2DM bolesnika imali euhromatične nukleu uz povećanje strukturnih praznina kod T2DM bolesnika. Nivo glukoze našte i HbA1c koreliraju sa fraktalnom dimenzijom i sa parametrima oblika nukleusa. Postoji korelacija između nivoa triglicerida u krvi i fraktalne dimenzije nukleusa.
Zaključak: Nukleusi limfocita bolesnika sa T2DM su nepravilnog oblika i sa većom količinom euhromatina, a promena njihovog izgleda je u vezi sa nivoom glikemije.",
publisher = "Beograd: Srpsko lekarsko društvo, Sekcija kliničke biohemije",
journal = "Knjiga sažetaka: 1. kongres kliničkih biohemičara i specijalista laboratorijske medicine Srbije sa međunarodnim učešćem;  2019 Nov 27-29; Belgrade, Serbia",
title = "Uticaj glikemije i lipidnog statusa na morfološke karakteristike nukleusa kod pacijenata sa tip 2 dijabetes melitusom",
pages = "58",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6662"
}

Martinović, T., Ćirić, D., Pantić, I., Lalić, K., Rasulić, I., Despotović, S., Lalić, I., Đuričić, D., Vučićević, L., Misirkić Marjanović, M., Trajković, V., Bumbaširević, V.,& Kravić-Stevović, T.. (2019). Uticaj glikemije i lipidnog statusa na morfološke karakteristike nukleusa kod pacijenata sa tip 2 dijabetes melitusom. in Knjiga sažetaka: 1. kongres kliničkih biohemičara i specijalista laboratorijske medicine Srbije sa međunarodnim učešćem;  2019 Nov 27-29; Belgrade, Serbia
Beograd: Srpsko lekarsko društvo, Sekcija kliničke biohemije., 58.
https://hdl.handle.net/21.15107/rcub_ibiss_6662

Martinović T, Ćirić D, Pantić I, Lalić K, Rasulić I, Despotović S, Lalić I, Đuričić D, Vučićević L, Misirkić Marjanović M, Trajković V, Bumbaširević V, Kravić-Stevović T. Uticaj glikemije i lipidnog statusa na morfološke karakteristike nukleusa kod pacijenata sa tip 2 dijabetes melitusom. in Knjiga sažetaka: 1. kongres kliničkih biohemičara i specijalista laboratorijske medicine Srbije sa međunarodnim učešćem;  2019 Nov 27-29; Belgrade, Serbia. 2019;:58.
https://hdl.handle.net/21.15107/rcub_ibiss_6662 .

Martinović, Tamara, Ćirić, Darko, Pantić, Igor, Lalić, Katarina, Rasulić, Iva, Despotović, Sanja, Lalić, Ivana, Đuričić, Danica, Vučićević, Ljubica, Misirkić Marjanović, Maja, Trajković, Vladimir, Bumbaširević, Vladimir, Kravić-Stevović, Tamara, "Uticaj glikemije i lipidnog statusa na morfološke karakteristike nukleusa kod pacijenata sa tip 2 dijabetes melitusom" in Knjiga sažetaka: 1. kongres kliničkih biohemičara i specijalista laboratorijske medicine Srbije sa međunarodnim učešćem;  2019 Nov 27-29; Belgrade, Serbia (2019):58,
https://hdl.handle.net/21.15107/rcub_ibiss_6662 .

TY  - CONF
AU  - Mandić, Miloš
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Jovanović, Maja
AU  - Bošnjak, Mihajlo
AU  - Perović, Vladimir
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2019
UR  - https://nordicautophagy.org
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6597
AB  - We investigated the mechanism and the role of autophagy in phorbol myristate acetate (PMA)-induced myeloid differentiation of human acute myeloid leukemia HL-60 cells. Methods: The mRNA levels of myeloid differentiation markers colony stimulating factor 1 receptor (CSF1R), early growth response protein 1 (EGR1), and interleukin 8 (IL-8), were assessed by real-time RT-PCR. Cell cycle arrest and the expression of surface myeloid marker CD11b were analyzed by flow cytometry. Autophagy was monitored by acridine orange staining, RT-PCR analysis of autophagy-related (ATG) gene expression, LC3-II/p62 immunoblotting, Beclin-1/Bcl-2 interaction, nuclear translocation of transcription factor EB (TFEB). The activation of MAP kinases extracelluar signal-regulated kinase (ERK) and c-Jun-N terminal kinase (JNK) was assessed by immunoblotting. Pharmacological inhibition and RNA interference (RNAi) were used to determine the role of MAP kinases in autophagy and HL60 cell differentiation, while the role of autophagy in HL60 differentiation was analyzed using RNAi-mediated knockdown of ATG5 and p62. Results: PMA-induced differentiation of HL-60 cells into macrophage-like cells was confirmed by cell-cycle arrest accompanied by elevated expression of p21, CD11b, CSF1R, EGR1, and IL-8. The induction of autophagy was demonstrated by accumulation of LC3-II, the increase in autophagic flux, the increase in expression of ATG genes, nuclear translocation of TFEB and dissociation of Beclin1from Bcl-2.The suppression of autophagy by RNAi–mediated knockdown of ATG5 or p62 counteracted myeloid differentiation of HL60 cells. Both ERK and JNK were activated by PMA, and their pharmacological and genetic inhibition decreased PMA-induced autophagy and differentiation of HL60 cells. Conclusion: Our study revealed the involvement of JNK and ERK in autophagy-dependent myeloid differentiation of HL60 cells, indicating MAP kinase-mediated autophagy as a possible target for treatment of acute myeloid leukemia
PB  - Nordic Autophagy Society
C3  - 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Netherlands
T1  - MAP kinase-dependent autophagy is involved in phorbol myristate acetate differentiation of HL-60 leukemia cells
SP  - 33
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6597
ER  - 

@conference{
author = "Mandić, Miloš and Misirkić Marjanović, Maja and Vučićević, Ljubica and Jovanović, Maja and Bošnjak, Mihajlo and Perović, Vladimir and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2019",
abstract = "We investigated the mechanism and the role of autophagy in phorbol myristate acetate (PMA)-induced myeloid differentiation of human acute myeloid leukemia HL-60 cells. Methods: The mRNA levels of myeloid differentiation markers colony stimulating factor 1 receptor (CSF1R), early growth response protein 1 (EGR1), and interleukin 8 (IL-8), were assessed by real-time RT-PCR. Cell cycle arrest and the expression of surface myeloid marker CD11b were analyzed by flow cytometry. Autophagy was monitored by acridine orange staining, RT-PCR analysis of autophagy-related (ATG) gene expression, LC3-II/p62 immunoblotting, Beclin-1/Bcl-2 interaction, nuclear translocation of transcription factor EB (TFEB). The activation of MAP kinases extracelluar signal-regulated kinase (ERK) and c-Jun-N terminal kinase (JNK) was assessed by immunoblotting. Pharmacological inhibition and RNA interference (RNAi) were used to determine the role of MAP kinases in autophagy and HL60 cell differentiation, while the role of autophagy in HL60 differentiation was analyzed using RNAi-mediated knockdown of ATG5 and p62. Results: PMA-induced differentiation of HL-60 cells into macrophage-like cells was confirmed by cell-cycle arrest accompanied by elevated expression of p21, CD11b, CSF1R, EGR1, and IL-8. The induction of autophagy was demonstrated by accumulation of LC3-II, the increase in autophagic flux, the increase in expression of ATG genes, nuclear translocation of TFEB and dissociation of Beclin1from Bcl-2.The suppression of autophagy by RNAi–mediated knockdown of ATG5 or p62 counteracted myeloid differentiation of HL60 cells. Both ERK and JNK were activated by PMA, and their pharmacological and genetic inhibition decreased PMA-induced autophagy and differentiation of HL60 cells. Conclusion: Our study revealed the involvement of JNK and ERK in autophagy-dependent myeloid differentiation of HL60 cells, indicating MAP kinase-mediated autophagy as a possible target for treatment of acute myeloid leukemia",
publisher = "Nordic Autophagy Society",
journal = "3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Netherlands",
title = "MAP kinase-dependent autophagy is involved in phorbol myristate acetate differentiation of HL-60 leukemia cells",
pages = "33",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6597"
}

Mandić, M., Misirkić Marjanović, M., Vučićević, L., Jovanović, M., Bošnjak, M., Perović, V., Harhaji-Trajković, L.,& Trajković, V.. (2019). MAP kinase-dependent autophagy is involved in phorbol myristate acetate differentiation of HL-60 leukemia cells. in 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Netherlands
Nordic Autophagy Society., 33.
https://hdl.handle.net/21.15107/rcub_ibiss_6597

Mandić M, Misirkić Marjanović M, Vučićević L, Jovanović M, Bošnjak M, Perović V, Harhaji-Trajković L, Trajković V. MAP kinase-dependent autophagy is involved in phorbol myristate acetate differentiation of HL-60 leukemia cells. in 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Netherlands. 2019;:33.
https://hdl.handle.net/21.15107/rcub_ibiss_6597 .

Mandić, Miloš, Misirkić Marjanović, Maja, Vučićević, Ljubica, Jovanović, Maja, Bošnjak, Mihajlo, Perović, Vladimir, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "MAP kinase-dependent autophagy is involved in phorbol myristate acetate differentiation of HL-60 leukemia cells" in 3rd Nordic Autophagy Society (NAS) Conference; 2019 May 22-24; Utrecht, Netherlands (2019):33,
https://hdl.handle.net/21.15107/rcub_ibiss_6597 .

TY  - CONF
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Kosić, Milica
AU  - Paunović, Verica
AU  - Arsikin-Csordas, Katarina
AU  - Ristić, Biljana
AU  - Marić, Nađa
AU  - Bošnjak, Mihajlo
AU  - Zogović, Nevena
AU  - Mandić, Miloš
AU  - Kravić-Stevović, Tamara
AU  - Martinović, Tamara
AU  - Ćirić, Darko
AU  - Mirčić, Aleksandar
AU  - Petričević, Saša
AU  - Bumbaširević, Vladimir
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2019
UR  - https://www.mitoeagle.org/index.php/MiP2019/MitoEAGLE_Belgrade_RS
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6353
AB  - We analyzed the impact of mitochondrial damage in anticancer action of combining lysosomal
membrane permeabilization (LMP)-inducing agent N- dodecylimidazole (NDI)[1] with
glycolytic inhibitor 2-deoxy-D-glucose (2DG) and in antipsychotic action of atypical antipsychotic
olanzapine.
NDI-triggered LMP and 2DG-mediated glycolysis block synergized in inducing ATP depletion,
mitochondrial damage and reactive oxygen species production, eventually leading to necrotic
death of U251 glioma cells but not primary astrocytes. NDI/2DG-induced death of glioma
cells was partly prevented by lysosomal cathepsin inhibitor E64 and antioxidant tocopherol, suggesting
the involvement of LMP and oxidative stress in the observed cytotoxicity. Moreover, the
combined oral administration of NDI and 2DG reduced in vivo melanoma growth in C57BL/6
mice by inducing necrotic death of tumor cells.
Based on these results, we propose that NDI-triggered LMPcauses initial mitochondrial damage
that is further increased by 2DG due to the lack of glycolytic ATP required to maintain mitochondrial
health. This leads to a positive feedback cycle of mitochondrial dysfunction, ATP loss,
and reactive oxygen species production, culminating in necrotic cell death.
We also investigated the role of autophagy, a controlled cellular self-digestion process, in regulating
survival of neurons exposed to olanzapine. Olanzapine induced autophagy in human
SH-SY5Y neuronal cell line, as confirmed by the increase in autophagic flux and presence of
autophagic vesicles, fusion of autophagosomes with lysosomes, and increase in the expression
of autophagy-related (ATG) genes ATG4B, ATG5, andATG7. The production of reactive oxygen
species, but not modulation of the main autophagy repressor mTOR or its upstream regulators
AMP-activated protein kinase and AKT1, was responsible for olanzapine-triggered autophagy.
Olanzapine-mediated oxidative stress also induced mitochondrial depolarization and damage,
and the autophagic clearance of dysfunctional mitochondria [2] was confirmed by electron microscopy,
colocalization of autophagosome associated MAP1LC3B (LC3B) and mitochondria,
and mitochondrial association with the autophagic cargo receptor p62. While olanzapine-triggered
mitochondrial damage was not visibly toxic to SH-SY5Ycells, their death was readily initiated
upon the inhibition of autophagy with pharmacological inhibitors, RNA interference knockdown
of BECN1 and LC3B. The treatment of mice with olanzapine increased the brain levels of
LC3B-II and mRNA encoding Atg4b,Atg5, Atg7, Atg12, Gabarap, and Becn1.
These data indicate that olanzapine-triggered autophagy protects neurons from otherwise fatal
mitochondrial damage, and that inhibition of autophagy might unmask the neurotoxic action
of the drug.
References;
1. Repnik U, Turk B (2010) Lysosomal-mitochondrial cross-talk during cell death.
Mitochondrion10: 662-669.
2. Wang K, Klionsky DJ(2011) Mitochondrial removal by autophagy. Autophagy 7:297-300.
PB  - The Mitochondrial Physiology Society
C3  - Programme abstract book: 14th Conference on Mitochondrial Physiology: Mitochondrial function: changes during life cycle and in noncommunicable diseases: COST MitoEAGLE perspectives and MitoEAGLE WG and MC Meeting: MiP2019/MitoEAGLE; 2019 Oct 13-16; Belgrade, Serbia
T1  - Dual role of mitochondrial damage in anticancer and antipsychotic treatment
SP  - 29
EP  - 29
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6353
ER  - 

@conference{
author = "Misirkić Marjanović, Maja and Vučićević, Ljubica and Kosić, Milica and Paunović, Verica and Arsikin-Csordas, Katarina and Ristić, Biljana and Marić, Nađa and Bošnjak, Mihajlo and Zogović, Nevena and Mandić, Miloš and Kravić-Stevović, Tamara and Martinović, Tamara and Ćirić, Darko and Mirčić, Aleksandar and Petričević, Saša and Bumbaširević, Vladimir and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2019",
abstract = "We analyzed the impact of mitochondrial damage in anticancer action of combining lysosomal
membrane permeabilization (LMP)-inducing agent N- dodecylimidazole (NDI)[1] with
glycolytic inhibitor 2-deoxy-D-glucose (2DG) and in antipsychotic action of atypical antipsychotic
olanzapine.
NDI-triggered LMP and 2DG-mediated glycolysis block synergized in inducing ATP depletion,
mitochondrial damage and reactive oxygen species production, eventually leading to necrotic
death of U251 glioma cells but not primary astrocytes. NDI/2DG-induced death of glioma
cells was partly prevented by lysosomal cathepsin inhibitor E64 and antioxidant tocopherol, suggesting
the involvement of LMP and oxidative stress in the observed cytotoxicity. Moreover, the
combined oral administration of NDI and 2DG reduced in vivo melanoma growth in C57BL/6
mice by inducing necrotic death of tumor cells.
Based on these results, we propose that NDI-triggered LMPcauses initial mitochondrial damage
that is further increased by 2DG due to the lack of glycolytic ATP required to maintain mitochondrial
health. This leads to a positive feedback cycle of mitochondrial dysfunction, ATP loss,
and reactive oxygen species production, culminating in necrotic cell death.
We also investigated the role of autophagy, a controlled cellular self-digestion process, in regulating
survival of neurons exposed to olanzapine. Olanzapine induced autophagy in human
SH-SY5Y neuronal cell line, as confirmed by the increase in autophagic flux and presence of
autophagic vesicles, fusion of autophagosomes with lysosomes, and increase in the expression
of autophagy-related (ATG) genes ATG4B, ATG5, andATG7. The production of reactive oxygen
species, but not modulation of the main autophagy repressor mTOR or its upstream regulators
AMP-activated protein kinase and AKT1, was responsible for olanzapine-triggered autophagy.
Olanzapine-mediated oxidative stress also induced mitochondrial depolarization and damage,
and the autophagic clearance of dysfunctional mitochondria [2] was confirmed by electron microscopy,
colocalization of autophagosome associated MAP1LC3B (LC3B) and mitochondria,
and mitochondrial association with the autophagic cargo receptor p62. While olanzapine-triggered
mitochondrial damage was not visibly toxic to SH-SY5Ycells, their death was readily initiated
upon the inhibition of autophagy with pharmacological inhibitors, RNA interference knockdown
of BECN1 and LC3B. The treatment of mice with olanzapine increased the brain levels of
LC3B-II and mRNA encoding Atg4b,Atg5, Atg7, Atg12, Gabarap, and Becn1.
These data indicate that olanzapine-triggered autophagy protects neurons from otherwise fatal
mitochondrial damage, and that inhibition of autophagy might unmask the neurotoxic action
of the drug.
References;
1. Repnik U, Turk B (2010) Lysosomal-mitochondrial cross-talk during cell death.
Mitochondrion10: 662-669.
2. Wang K, Klionsky DJ(2011) Mitochondrial removal by autophagy. Autophagy 7:297-300.",
publisher = "The Mitochondrial Physiology Society",
journal = "Programme abstract book: 14th Conference on Mitochondrial Physiology: Mitochondrial function: changes during life cycle and in noncommunicable diseases: COST MitoEAGLE perspectives and MitoEAGLE WG and MC Meeting: MiP2019/MitoEAGLE; 2019 Oct 13-16; Belgrade, Serbia",
title = "Dual role of mitochondrial damage in anticancer and antipsychotic treatment",
pages = "29-29",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6353"
}

Misirkić Marjanović, M., Vučićević, L., Kosić, M., Paunović, V., Arsikin-Csordas, K., Ristić, B., Marić, N., Bošnjak, M., Zogović, N., Mandić, M., Kravić-Stevović, T., Martinović, T., Ćirić, D., Mirčić, A., Petričević, S., Bumbaširević, V., Harhaji-Trajković, L.,& Trajković, V.. (2019). Dual role of mitochondrial damage in anticancer and antipsychotic treatment. in Programme abstract book: 14th Conference on Mitochondrial Physiology: Mitochondrial function: changes during life cycle and in noncommunicable diseases: COST MitoEAGLE perspectives and MitoEAGLE WG and MC Meeting: MiP2019/MitoEAGLE; 2019 Oct 13-16; Belgrade, Serbia
The Mitochondrial Physiology Society., 29-29.
https://hdl.handle.net/21.15107/rcub_ibiss_6353

Misirkić Marjanović M, Vučićević L, Kosić M, Paunović V, Arsikin-Csordas K, Ristić B, Marić N, Bošnjak M, Zogović N, Mandić M, Kravić-Stevović T, Martinović T, Ćirić D, Mirčić A, Petričević S, Bumbaširević V, Harhaji-Trajković L, Trajković V. Dual role of mitochondrial damage in anticancer and antipsychotic treatment. in Programme abstract book: 14th Conference on Mitochondrial Physiology: Mitochondrial function: changes during life cycle and in noncommunicable diseases: COST MitoEAGLE perspectives and MitoEAGLE WG and MC Meeting: MiP2019/MitoEAGLE; 2019 Oct 13-16; Belgrade, Serbia. 2019;:29-29.
https://hdl.handle.net/21.15107/rcub_ibiss_6353 .

Misirkić Marjanović, Maja, Vučićević, Ljubica, Kosić, Milica, Paunović, Verica, Arsikin-Csordas, Katarina, Ristić, Biljana, Marić, Nađa, Bošnjak, Mihajlo, Zogović, Nevena, Mandić, Miloš, Kravić-Stevović, Tamara, Martinović, Tamara, Ćirić, Darko, Mirčić, Aleksandar, Petričević, Saša, Bumbaširević, Vladimir, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "Dual role of mitochondrial damage in anticancer and antipsychotic treatment" in Programme abstract book: 14th Conference on Mitochondrial Physiology: Mitochondrial function: changes during life cycle and in noncommunicable diseases: COST MitoEAGLE perspectives and MitoEAGLE WG and MC Meeting: MiP2019/MitoEAGLE; 2019 Oct 13-16; Belgrade, Serbia (2019):29-29,
https://hdl.handle.net/21.15107/rcub_ibiss_6353 .

TY  - CONF
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Ćirić, Darko
AU  - Martinović, Tamara
AU  - Jovanović, Maja
AU  - Isaković, Aleksandra
AU  - Marković, Ivanka
AU  - Zogović, Nevena
AU  - Foretz, Mark
AU  - Rabanal-Ruiz, Yoana
AU  - Korolchuk, Viktor
AU  - Trajković, Vladimir
PY  - 2019
UR  - https://www.fens.org/news-activities/fens-and-societies-calendar/meeting-event/fens-regional-meeting-2019
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6357
AB  - We investigated the effect of excitotoxic glutamate on nutrient starvation-induced autophagy, a process of lysosome-mediated degradation of cellular macromolecules and organelles. Incubation of SH-SY5Y human neuroblastoma cell line in glucose/amino acid/serum-free Hank Balanced Salt solution synergized with glutamate in causing energy stress and excitotoxic necrosis. Glutamate inhibited starvation-induced autophagy, as demonstrated by decreased intracellular acidification, lower LC3 punctuation, reduced conversion of LC3-I to LC3-II, reduced expression of autophagy activators beclin-1 and ATG5, increased
levels of the selective autophagic target NBR1, and decline in the number of autophagic vesicles observed by transmission electron microscopy. NMDA antagonist memantine restored LC3B-II accumulation in starved cells exposed to glutamate, indicating that glutamate exerts its inhibitory role on autophagy by activating NMDA receptors. The modulation of mTOR, the negative regulator of autophagy, was not responsible for glutamate-mediated autophagy inhibition during starvation. On the other hand, glutamate downregulated starvation-induced activation of the intracellular energy sensor AMP-activated protein
kinase (AMPK). This was associated with reduced mRNA expression of autophagy transcription factors FOXO3 and ATF4, as well as molecules involved in autophagy process (ULK1, ATG13, FIP200, ATG14, beclin-1, ATG5, ATG12, SQSTM1). The ability of glutamate to repress transcription of autophagy genes in starved cells was partly mediated by AMPK downregulation. Genetic or pharmacological AMPK activation by AMPK overexpression or metformin, as well as genetic or pharmacological autophagy induction by TFEB overexpression or lithium chloride, rescued cells from glutamate-mediated excitoxicity. These data indicate that transcriptional inhibition of AMPK-dependent autophagy is involved in glutamate-mediated excitotoxicity during nutrient deprivation in vitro.
PB  - Belgrade : Serbian Neuroscience Society
C3  - Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia
T1  - Autophagy regulation and its role in glutamate excitotoxicity during nutrient stress
SP  - 144
EP  - 144
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6357
ER  - 

@conference{
author = "Vučićević, Ljubica and Misirkić Marjanović, Maja and Ćirić, Darko and Martinović, Tamara and Jovanović, Maja and Isaković, Aleksandra and Marković, Ivanka and Zogović, Nevena and Foretz, Mark and Rabanal-Ruiz, Yoana and Korolchuk, Viktor and Trajković, Vladimir",
year = "2019",
abstract = "We investigated the effect of excitotoxic glutamate on nutrient starvation-induced autophagy, a process of lysosome-mediated degradation of cellular macromolecules and organelles. Incubation of SH-SY5Y human neuroblastoma cell line in glucose/amino acid/serum-free Hank Balanced Salt solution synergized with glutamate in causing energy stress and excitotoxic necrosis. Glutamate inhibited starvation-induced autophagy, as demonstrated by decreased intracellular acidification, lower LC3 punctuation, reduced conversion of LC3-I to LC3-II, reduced expression of autophagy activators beclin-1 and ATG5, increased
levels of the selective autophagic target NBR1, and decline in the number of autophagic vesicles observed by transmission electron microscopy. NMDA antagonist memantine restored LC3B-II accumulation in starved cells exposed to glutamate, indicating that glutamate exerts its inhibitory role on autophagy by activating NMDA receptors. The modulation of mTOR, the negative regulator of autophagy, was not responsible for glutamate-mediated autophagy inhibition during starvation. On the other hand, glutamate downregulated starvation-induced activation of the intracellular energy sensor AMP-activated protein
kinase (AMPK). This was associated with reduced mRNA expression of autophagy transcription factors FOXO3 and ATF4, as well as molecules involved in autophagy process (ULK1, ATG13, FIP200, ATG14, beclin-1, ATG5, ATG12, SQSTM1). The ability of glutamate to repress transcription of autophagy genes in starved cells was partly mediated by AMPK downregulation. Genetic or pharmacological AMPK activation by AMPK overexpression or metformin, as well as genetic or pharmacological autophagy induction by TFEB overexpression or lithium chloride, rescued cells from glutamate-mediated excitoxicity. These data indicate that transcriptional inhibition of AMPK-dependent autophagy is involved in glutamate-mediated excitotoxicity during nutrient deprivation in vitro.",
publisher = "Belgrade : Serbian Neuroscience Society",
journal = "Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia",
title = "Autophagy regulation and its role in glutamate excitotoxicity during nutrient stress",
pages = "144-144",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6357"
}

Vučićević, L., Misirkić Marjanović, M., Ćirić, D., Martinović, T., Jovanović, M., Isaković, A., Marković, I., Zogović, N., Foretz, M., Rabanal-Ruiz, Y., Korolchuk, V.,& Trajković, V.. (2019). Autophagy regulation and its role in glutamate excitotoxicity during nutrient stress. in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia
Belgrade : Serbian Neuroscience Society., 144-144.
https://hdl.handle.net/21.15107/rcub_ibiss_6357

Vučićević L, Misirkić Marjanović M, Ćirić D, Martinović T, Jovanović M, Isaković A, Marković I, Zogović N, Foretz M, Rabanal-Ruiz Y, Korolchuk V, Trajković V. Autophagy regulation and its role in glutamate excitotoxicity during nutrient stress. in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia. 2019;:144-144.
https://hdl.handle.net/21.15107/rcub_ibiss_6357 .

Vučićević, Ljubica, Misirkić Marjanović, Maja, Ćirić, Darko, Martinović, Tamara, Jovanović, Maja, Isaković, Aleksandra, Marković, Ivanka, Zogović, Nevena, Foretz, Mark, Rabanal-Ruiz, Yoana, Korolchuk, Viktor, Trajković, Vladimir, "Autophagy regulation and its role in glutamate excitotoxicity during nutrient stress" in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia (2019):144-144,
https://hdl.handle.net/21.15107/rcub_ibiss_6357 .

TY  - CONF
AU  - Mandić, Miloš
AU  - Vučićević, Ljubica
AU  - Misirkić Marjanović, Maja
AU  - Jovanović, Maja
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2018
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6287
AB  - Introduction: Phorbol myristate acetate (PMA) is in
clinical investigation for treatment of acute myeloid
leukemia due to its differentiating ability. Cell differentiation could be accompanied by senescence, a state
of irreversible cell cycle arrest.
Our aim was to investigate the ability of PMA to initiate
senescence in HL60 human leukemia cells.
Methods: Cell morphology was analyzed using phase
contrast microscopy. Cell cycle arrest was assessed
by flow cytometric analysis of propidium iodide stained
cells and BrdU colorimetric assay. Activity of senescence-associated beta-galactosidase (SA-βgal) was
assessed by cytochemical staining and flow cytometric analysis of fluorescein di-β-D-galactopyranoside
(FDG) stained cells. Senescence-associated gene expression of: cell cycle inhibitor p21, interleukin-8 (IL-8),
lamin B1 were quantified by RT-PCR, while activation
of NF-κB, main regulator of senescence associated secretory phenotype, was examined by immunoblotting.
Results: After the PMA treatment HL60 were enlarged and flattened with cytoplasmic vacuoles resembling morphology of senescent cells. Block in
leukemia cell proliferation in G1 phase was accompanied with increase in expression of cell cycle inhibitor p21 in PMA treated cells. Finally, PMA stimulated
SA-βgal activity, expression of genes responsible for
senescence associated secretory phenotype, NF-κB
and IL-8, while downregulating Lamin B1 expression.
Conclusion: Our results suggest that in addition to
PMA-induced cellular differentiation, senescence
participates in its previously shown cytostatic effect,
further supporting its investigation as a potential antileukemic drug.
PB  - Belgrade: Institute for Biological Research "Siniša Stanković", University of Belgrade
C3  - Program & Book of Abstracts. IUBMB Advanced School Nutrition, Metabolism and Aging; 2018 Oct 15-19; Petnica, Serbia
T1  - Phorbol 12-myristate 13-acetate induces senescence of HL-60 leukemic cells
SP  - 38
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6287
ER  - 

@conference{
author = "Mandić, Miloš and Vučićević, Ljubica and Misirkić Marjanović, Maja and Jovanović, Maja and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2018",
abstract = "Introduction: Phorbol myristate acetate (PMA) is in
clinical investigation for treatment of acute myeloid
leukemia due to its differentiating ability. Cell differentiation could be accompanied by senescence, a state
of irreversible cell cycle arrest.
Our aim was to investigate the ability of PMA to initiate
senescence in HL60 human leukemia cells.
Methods: Cell morphology was analyzed using phase
contrast microscopy. Cell cycle arrest was assessed
by flow cytometric analysis of propidium iodide stained
cells and BrdU colorimetric assay. Activity of senescence-associated beta-galactosidase (SA-βgal) was
assessed by cytochemical staining and flow cytometric analysis of fluorescein di-β-D-galactopyranoside
(FDG) stained cells. Senescence-associated gene expression of: cell cycle inhibitor p21, interleukin-8 (IL-8),
lamin B1 were quantified by RT-PCR, while activation
of NF-κB, main regulator of senescence associated secretory phenotype, was examined by immunoblotting.
Results: After the PMA treatment HL60 were enlarged and flattened with cytoplasmic vacuoles resembling morphology of senescent cells. Block in
leukemia cell proliferation in G1 phase was accompanied with increase in expression of cell cycle inhibitor p21 in PMA treated cells. Finally, PMA stimulated
SA-βgal activity, expression of genes responsible for
senescence associated secretory phenotype, NF-κB
and IL-8, while downregulating Lamin B1 expression.
Conclusion: Our results suggest that in addition to
PMA-induced cellular differentiation, senescence
participates in its previously shown cytostatic effect,
further supporting its investigation as a potential antileukemic drug.",
publisher = "Belgrade: Institute for Biological Research "Siniša Stanković", University of Belgrade",
journal = "Program & Book of Abstracts. IUBMB Advanced School Nutrition, Metabolism and Aging; 2018 Oct 15-19; Petnica, Serbia",
title = "Phorbol 12-myristate 13-acetate induces senescence of HL-60 leukemic cells",
pages = "38",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6287"
}

Mandić, M., Vučićević, L., Misirkić Marjanović, M., Jovanović, M., Harhaji-Trajković, L.,& Trajković, V.. (2018). Phorbol 12-myristate 13-acetate induces senescence of HL-60 leukemic cells. in Program & Book of Abstracts. IUBMB Advanced School Nutrition, Metabolism and Aging; 2018 Oct 15-19; Petnica, Serbia
Belgrade: Institute for Biological Research "Siniša Stanković", University of Belgrade., 38.
https://hdl.handle.net/21.15107/rcub_ibiss_6287

Mandić M, Vučićević L, Misirkić Marjanović M, Jovanović M, Harhaji-Trajković L, Trajković V. Phorbol 12-myristate 13-acetate induces senescence of HL-60 leukemic cells. in Program & Book of Abstracts. IUBMB Advanced School Nutrition, Metabolism and Aging; 2018 Oct 15-19; Petnica, Serbia. 2018;:38.
https://hdl.handle.net/21.15107/rcub_ibiss_6287 .

Mandić, Miloš, Vučićević, Ljubica, Misirkić Marjanović, Maja, Jovanović, Maja, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "Phorbol 12-myristate 13-acetate induces senescence of HL-60 leukemic cells" in Program & Book of Abstracts. IUBMB Advanced School Nutrition, Metabolism and Aging; 2018 Oct 15-19; Petnica, Serbia (2018):38,
https://hdl.handle.net/21.15107/rcub_ibiss_6287 .

TY  - GEN
AU  - Gazdić, Marina
AU  - Simović Marković, Bojana
AU  - Vučićević, Ljubica
AU  - Nikolić, Tamara
AU  - Đonov, Valentin
AU  - Arsenijević, Nebojša
AU  - Trajković, Vladimir
AU  - Lukić, Miodrag L.
AU  - Volarević, Vladislav
PY  - 2018
UR  - http://doi.wiley.com/10.1002/term.2452
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2833
AB  - The effects of mesenchymal stem cells (MSCs) on the phenotype and function of natural killer T (NKT) cells is not understood. We used concanavalin A (Con A) and α-galactosylceramide (α-GalCer)-induced liver injury to evaluate the effects of MSCs on NKT-dependent hepatotoxicity. Mouse MSCs (mMSCs) significantly reduced Con A- and α-GalCer-mediated hepatitis in C57Bl/6 mice, as demonstrated by histopathological and biochemical analysis, attenuated the influx of inflammatory [T-bet + , tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ)-producing and GATA3 + , interleukin-4 (IL-4)-producing] liver NKT cells and downregulated TNF-α, IFN-γ and IL-4 levels in the sera. The liver NKT cells cultured in vitro with mMSCs produced lower amounts of inflammatory cytokines (TNF-α, IFN-γ, IL-4) and higher amounts of immunosuppressive IL-10 upon α-GalCer stimulation. mMSC treatment attenuated expression of apoptosis-inducing ligands on liver NKT cells and suppressed the expression of pro-apoptotic genes in the livers of α-GalCer-treated mice. mMSCs reduced the cytotoxicity of liver NKT cells against hepatocytes in vitro. The presence of 1-methyl-dl-tryptophan, a specific inhibitor of indoleamine 2,3-dioxygenase (IDO), or l-N G -monomethyl arginine citrate, a specific inhibitor of inducible nitric oxide synthase (iNOS), in mMSC-conditioned medium injected into α-GalCer-treated mice, counteracted the hepatoprotective effect of mMSCs in vivo and restored pro-inflammatory cytokine production and cytotoxicity of NKT cells in vitro. Human MSCs attenuated the production of inflammatory cytokines in α-GalCer-stimulated human peripheral blood mononuclear cells in an iNOS- and IDO-dependent manner and reduced their cytotoxicity against HepG2 cells. In conclusion, MSCs protect from acute liver injury by attenuating the cytotoxicity and capacity of liver NKT cells to produce inflammatory cytokines in an iNOS- and IDO-dependent manner.
T2  - Journal of Tissue Engineering and Regenerative Medicine
T1  - Mesenchymal stem cells protect from acute liver injury by attenuating hepatotoxicity of liver natural killer T cells in an inducible nitric oxide synthase- and indoleamine 2,3-dioxygenase-dependent manner
IS  - 2
VL  - 12
DO  - 10.1002/term.2452
SP  - e1173
EP  - e1185
ER  - 

@misc{
author = "Gazdić, Marina and Simović Marković, Bojana and Vučićević, Ljubica and Nikolić, Tamara and Đonov, Valentin and Arsenijević, Nebojša and Trajković, Vladimir and Lukić, Miodrag L. and Volarević, Vladislav",
year = "2018",
abstract = "The effects of mesenchymal stem cells (MSCs) on the phenotype and function of natural killer T (NKT) cells is not understood. We used concanavalin A (Con A) and α-galactosylceramide (α-GalCer)-induced liver injury to evaluate the effects of MSCs on NKT-dependent hepatotoxicity. Mouse MSCs (mMSCs) significantly reduced Con A- and α-GalCer-mediated hepatitis in C57Bl/6 mice, as demonstrated by histopathological and biochemical analysis, attenuated the influx of inflammatory [T-bet + , tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ)-producing and GATA3 + , interleukin-4 (IL-4)-producing] liver NKT cells and downregulated TNF-α, IFN-γ and IL-4 levels in the sera. The liver NKT cells cultured in vitro with mMSCs produced lower amounts of inflammatory cytokines (TNF-α, IFN-γ, IL-4) and higher amounts of immunosuppressive IL-10 upon α-GalCer stimulation. mMSC treatment attenuated expression of apoptosis-inducing ligands on liver NKT cells and suppressed the expression of pro-apoptotic genes in the livers of α-GalCer-treated mice. mMSCs reduced the cytotoxicity of liver NKT cells against hepatocytes in vitro. The presence of 1-methyl-dl-tryptophan, a specific inhibitor of indoleamine 2,3-dioxygenase (IDO), or l-N G -monomethyl arginine citrate, a specific inhibitor of inducible nitric oxide synthase (iNOS), in mMSC-conditioned medium injected into α-GalCer-treated mice, counteracted the hepatoprotective effect of mMSCs in vivo and restored pro-inflammatory cytokine production and cytotoxicity of NKT cells in vitro. Human MSCs attenuated the production of inflammatory cytokines in α-GalCer-stimulated human peripheral blood mononuclear cells in an iNOS- and IDO-dependent manner and reduced their cytotoxicity against HepG2 cells. In conclusion, MSCs protect from acute liver injury by attenuating the cytotoxicity and capacity of liver NKT cells to produce inflammatory cytokines in an iNOS- and IDO-dependent manner.",
journal = "Journal of Tissue Engineering and Regenerative Medicine",
title = "Mesenchymal stem cells protect from acute liver injury by attenuating hepatotoxicity of liver natural killer T cells in an inducible nitric oxide synthase- and indoleamine 2,3-dioxygenase-dependent manner",
number = "2",
volume = "12",
doi = "10.1002/term.2452",
pages = "e1173-e1185"
}

Gazdić, M., Simović Marković, B., Vučićević, L., Nikolić, T., Đonov, V., Arsenijević, N., Trajković, V., Lukić, M. L.,& Volarević, V.. (2018). Mesenchymal stem cells protect from acute liver injury by attenuating hepatotoxicity of liver natural killer T cells in an inducible nitric oxide synthase- and indoleamine 2,3-dioxygenase-dependent manner. in Journal of Tissue Engineering and Regenerative Medicine, 12(2), e1173-e1185.
https://doi.org/10.1002/term.2452

Gazdić M, Simović Marković B, Vučićević L, Nikolić T, Đonov V, Arsenijević N, Trajković V, Lukić ML, Volarević V. Mesenchymal stem cells protect from acute liver injury by attenuating hepatotoxicity of liver natural killer T cells in an inducible nitric oxide synthase- and indoleamine 2,3-dioxygenase-dependent manner. in Journal of Tissue Engineering and Regenerative Medicine. 2018;12(2):e1173-e1185.
doi:10.1002/term.2452 .

Gazdić, Marina, Simović Marković, Bojana, Vučićević, Ljubica, Nikolić, Tamara, Đonov, Valentin, Arsenijević, Nebojša, Trajković, Vladimir, Lukić, Miodrag L., Volarević, Vladislav, "Mesenchymal stem cells protect from acute liver injury by attenuating hepatotoxicity of liver natural killer T cells in an inducible nitric oxide synthase- and indoleamine 2,3-dioxygenase-dependent manner" in Journal of Tissue Engineering and Regenerative Medicine, 12, no. 2 (2018):e1173-e1185,
https://doi.org/10.1002/term.2452 . .

TY  - JOUR
AU  - Zogović, Nevena
AU  - Tovilović-Kovačević, Gordana
AU  - Misirkić Marjanović, Maja
AU  - Vučićević, Ljubica
AU  - Janjetović, Kristina
AU  - Harhaji-Trajković, Ljubica
AU  - Trajkovic, Vladimir
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1978
AB  - We explored the interplay between the intracellular energy sensor
   AMP-activated protein kinase (AMPK), extracellular signal-regulated
   kinase (ERK), and autophagy in phorbol myristate acetate (PMA)-induced
   neuronal differentiation of SH-SY5Y human neuroblastoma cells.
   PMA-triggered expression of neuronal markers (dopamine transporter,
   microtubule-associated protein 2, -tubulin) was associated with an
   autophagic response, measured by the conversion of
   microtubule-associated protein light chain 3 (LC3)-I to
   autophagosome-bound LC3-II, increase in autophagic flux, and expression
   of autophagy-related (Atg) proteins Atg7 and beclin-1. This coincided
   with the transient activation of AMPK and sustained activation of ERK.
   Pharmacological inhibition or RNA interference-mediated silencing of
   AMPK suppressed PMA-induced expression of neuronal markers, as well as
   ERK activation and autophagy. A selective pharmacological blockade of
   ERK prevented PMA-induced neuronal differentiation and autophagy
   induction without affecting AMPK phosphorylation. Conversely, the
   inhibition of autophagy downstream of AMPK/ERK, either by
   pharmacological agents or LC3 knockdown, promoted the expression of
   neuronal markers, thus indicating a role of autophagy in the suppression
   of PMA-induced differentiation of SH-SY5Y cells. Therefore, PMA-induced
   neuronal differentiation of SH-SY5Y cells depends on a complex interplay
   between AMPK, ERK, and autophagy, in which the stimulatory effects of
   AMPK/ERK signaling are counteracted by the coinciding autophagic
   response.
T2  - Journal of Neurochemistry
T1  - Coordinated activation of AMP-activated protein kinase, extracellular
 signal-regulated kinase, and autophagy regulates phorbol myristate
 acetate-induced differentiation of SH-SY5Y neuroblastoma cells
IS  - 2
VL  - 133
DO  - 10.1111/jnc.12980
SP  - 223
EP  - 232
ER  - 

@article{
author = "Zogović, Nevena and Tovilović-Kovačević, Gordana and Misirkić Marjanović, Maja and Vučićević, Ljubica and Janjetović, Kristina and Harhaji-Trajković, Ljubica and Trajkovic, Vladimir",
year = "2015",
abstract = "We explored the interplay between the intracellular energy sensor
   AMP-activated protein kinase (AMPK), extracellular signal-regulated
   kinase (ERK), and autophagy in phorbol myristate acetate (PMA)-induced
   neuronal differentiation of SH-SY5Y human neuroblastoma cells.
   PMA-triggered expression of neuronal markers (dopamine transporter,
   microtubule-associated protein 2, -tubulin) was associated with an
   autophagic response, measured by the conversion of
   microtubule-associated protein light chain 3 (LC3)-I to
   autophagosome-bound LC3-II, increase in autophagic flux, and expression
   of autophagy-related (Atg) proteins Atg7 and beclin-1. This coincided
   with the transient activation of AMPK and sustained activation of ERK.
   Pharmacological inhibition or RNA interference-mediated silencing of
   AMPK suppressed PMA-induced expression of neuronal markers, as well as
   ERK activation and autophagy. A selective pharmacological blockade of
   ERK prevented PMA-induced neuronal differentiation and autophagy
   induction without affecting AMPK phosphorylation. Conversely, the
   inhibition of autophagy downstream of AMPK/ERK, either by
   pharmacological agents or LC3 knockdown, promoted the expression of
   neuronal markers, thus indicating a role of autophagy in the suppression
   of PMA-induced differentiation of SH-SY5Y cells. Therefore, PMA-induced
   neuronal differentiation of SH-SY5Y cells depends on a complex interplay
   between AMPK, ERK, and autophagy, in which the stimulatory effects of
   AMPK/ERK signaling are counteracted by the coinciding autophagic
   response.",
journal = "Journal of Neurochemistry",
title = "Coordinated activation of AMP-activated protein kinase, extracellular
 signal-regulated kinase, and autophagy regulates phorbol myristate
 acetate-induced differentiation of SH-SY5Y neuroblastoma cells",
number = "2",
volume = "133",
doi = "10.1111/jnc.12980",
pages = "223-232"
}

Zogović, N., Tovilović-Kovačević, G., Misirkić Marjanović, M., Vučićević, L., Janjetović, K., Harhaji-Trajković, L.,& Trajkovic, V.. (2015). Coordinated activation of AMP-activated protein kinase, extracellular
 signal-regulated kinase, and autophagy regulates phorbol myristate
 acetate-induced differentiation of SH-SY5Y neuroblastoma cells. in Journal of Neurochemistry, 133(2), 223-232.
https://doi.org/10.1111/jnc.12980

Zogović N, Tovilović-Kovačević G, Misirkić Marjanović M, Vučićević L, Janjetović K, Harhaji-Trajković L, Trajkovic V. Coordinated activation of AMP-activated protein kinase, extracellular
 signal-regulated kinase, and autophagy regulates phorbol myristate
 acetate-induced differentiation of SH-SY5Y neuroblastoma cells. in Journal of Neurochemistry. 2015;133(2):223-232.
doi:10.1111/jnc.12980 .

Zogović, Nevena, Tovilović-Kovačević, Gordana, Misirkić Marjanović, Maja, Vučićević, Ljubica, Janjetović, Kristina, Harhaji-Trajković, Ljubica, Trajkovic, Vladimir, "Coordinated activation of AMP-activated protein kinase, extracellular
 signal-regulated kinase, and autophagy regulates phorbol myristate
 acetate-induced differentiation of SH-SY5Y neuroblastoma cells" in Journal of Neurochemistry, 133, no. 2 (2015):223-232,
https://doi.org/10.1111/jnc.12980 . .

TY  - JOUR
AU  - Milovanovic, E. Sudar
AU  - Jovanovic, A.
AU  - Misirkic-Marjanovic, M.
AU  - Vučićević, Ljubica
AU  - Janjetović, Kristina
AU  - Isenovic, E. R.
PY  - 2015
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2342
AB  - The aim of this study was to examine the effects of ghrelin on
   regulation of cardiac inducible nitric oxide synthase (iNOS)
   activity/expression in high fat (HF), obese rats.
   For this study, male Wistar rats fed with HF diet (30 \% fat) for 4
   weeks were injected every 24 h for 5 days intracerebroventriculary (ICV)
   with ghrelin (0.3 nmol/5 mu l) or with an equal volume of phosphate
   buffered saline (PBS). Control rats were ICV injected with an equal
   volume of PBS. Glucose, insulin and nitric oxide (NO) concentrations
   were measured in serum, while arginase activity and citrulline
   concentrations were measured in heart lysate. Protein iNOS and
   regulatory subunit of nuclear factor-kappa B (NF kappa B-p65),
   phosphorylation of enzymes protein kinase B (Akt) at Ser(473), and
   extracellular signal-regulated kinases 1/2 (ERK1/2) at Tyr(202)/Tyr(204)
   were determined in heart lysate by Western blot. For gene expression of
   iNOS qRT-PCR was used.
   Results show significantly (p < 0.01) higher serum NO production in
   ghrelin treated HF rats compared with HF rats. Ghrelin significantly
   reduced citrulline concentration (p < 0.05) and arginase activity (p <
   0.01) in HF rats. In ghrelin treated HF rats, gene and protein
   expression of iNOS and NF kappa B-p65 levels were significantly (p <
   0.05) increased compared with HF rats. Increased phosphorylation of Akt
   (p < 0.01) and decreased (p < 0.05) ERK1/2 phosphorylation were detected
   in HF ghrelin treated rats compared with HF rats hearts.
   Results from this study indicate that exogenous ghrelin induces
   expression and activity of cardiac iNOS via Akt phosphorylation followed
   by NF kappa B activation in HF rats.
T2  - Experimental and Clinical Endocrinology & Diabetes
T1  - Effects of Intracerebroventricularly (ICV) Injected Ghrelin on Cardiac
 Inducible Nitric Oxide Synthase Activity/Expression in Obese Rats
IS  - 10
VL  - 123
DO  - 10.1055/s-0035-1559758
SP  - 581
EP  - 588
ER  - 

@article{
author = "Milovanovic, E. Sudar and Jovanovic, A. and Misirkic-Marjanovic, M. and Vučićević, Ljubica and Janjetović, Kristina and Isenovic, E. R.",
year = "2015",
abstract = "The aim of this study was to examine the effects of ghrelin on
   regulation of cardiac inducible nitric oxide synthase (iNOS)
   activity/expression in high fat (HF), obese rats.
   For this study, male Wistar rats fed with HF diet (30 \% fat) for 4
   weeks were injected every 24 h for 5 days intracerebroventriculary (ICV)
   with ghrelin (0.3 nmol/5 mu l) or with an equal volume of phosphate
   buffered saline (PBS). Control rats were ICV injected with an equal
   volume of PBS. Glucose, insulin and nitric oxide (NO) concentrations
   were measured in serum, while arginase activity and citrulline
   concentrations were measured in heart lysate. Protein iNOS and
   regulatory subunit of nuclear factor-kappa B (NF kappa B-p65),
   phosphorylation of enzymes protein kinase B (Akt) at Ser(473), and
   extracellular signal-regulated kinases 1/2 (ERK1/2) at Tyr(202)/Tyr(204)
   were determined in heart lysate by Western blot. For gene expression of
   iNOS qRT-PCR was used.
   Results show significantly (p < 0.01) higher serum NO production in
   ghrelin treated HF rats compared with HF rats. Ghrelin significantly
   reduced citrulline concentration (p < 0.05) and arginase activity (p <
   0.01) in HF rats. In ghrelin treated HF rats, gene and protein
   expression of iNOS and NF kappa B-p65 levels were significantly (p <
   0.05) increased compared with HF rats. Increased phosphorylation of Akt
   (p < 0.01) and decreased (p < 0.05) ERK1/2 phosphorylation were detected
   in HF ghrelin treated rats compared with HF rats hearts.
   Results from this study indicate that exogenous ghrelin induces
   expression and activity of cardiac iNOS via Akt phosphorylation followed
   by NF kappa B activation in HF rats.",
journal = "Experimental and Clinical Endocrinology & Diabetes",
title = "Effects of Intracerebroventricularly (ICV) Injected Ghrelin on Cardiac
 Inducible Nitric Oxide Synthase Activity/Expression in Obese Rats",
number = "10",
volume = "123",
doi = "10.1055/s-0035-1559758",
pages = "581-588"
}

Milovanovic, E. S., Jovanovic, A., Misirkic-Marjanovic, M., Vučićević, L., Janjetović, K.,& Isenovic, E. R.. (2015). Effects of Intracerebroventricularly (ICV) Injected Ghrelin on Cardiac
 Inducible Nitric Oxide Synthase Activity/Expression in Obese Rats. in Experimental and Clinical Endocrinology & Diabetes, 123(10), 581-588.
https://doi.org/10.1055/s-0035-1559758

Milovanovic ES, Jovanovic A, Misirkic-Marjanovic M, Vučićević L, Janjetović K, Isenovic ER. Effects of Intracerebroventricularly (ICV) Injected Ghrelin on Cardiac
 Inducible Nitric Oxide Synthase Activity/Expression in Obese Rats. in Experimental and Clinical Endocrinology & Diabetes. 2015;123(10):581-588.
doi:10.1055/s-0035-1559758 .

Milovanovic, E. Sudar, Jovanovic, A., Misirkic-Marjanovic, M., Vučićević, Ljubica, Janjetović, Kristina, Isenovic, E. R., "Effects of Intracerebroventricularly (ICV) Injected Ghrelin on Cardiac
 Inducible Nitric Oxide Synthase Activity/Expression in Obese Rats" in Experimental and Clinical Endocrinology & Diabetes, 123, no. 10 (2015):581-588,
https://doi.org/10.1055/s-0035-1559758 . .

TY  - JOUR
AU  - Tovilović-Kovačević, Gordana
AU  - Zogović, Nevena
AU  - Šoškić, Vukić
AU  - Schrattenholz, Andre
AU  - Kostić-Rajačić, Slađana
AU  - Misirkić Marjanović, Maja
AU  - Janjetović, Kristina
AU  - Vučićević, Ljubica
AU  - Arsikin, Katarina
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2013
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6343
AB  - We investigated the ability of 19 recently synthesized arylpiperazine compounds to protect human SHSY5Y neuroblastoma cells from the neurotoxin 6-hydroxydopamine (6-OHDA). The compound with the most potent neuroprotective action was N-{3-[2-(4-phenyl-piperazin-1-yl)-ethyl]-phenyl}-picolinamide (6b), which reduced 6-OHDA-induced apoptotic death through stabilization of mitochondrial membrane and subsequent prevention of superoxide production, caspase activation and DNA fragmentation. 6-OHDA-triggered autophagic response was also reduced by 6b, which prevented inactivation of the main autophagy repressor mTOR, upregulation of proautophagic beclin-1, conversion of microtubule-associated protein 1 light chain 3 (LC3)-I to autophagosome-associated LC3-II, as well as intracytoplasmic acidification induced by 6-OHDA. The inhibition of autophagy using LC3b gene silencing or pharmacological autophagy blockers 3-methyladenine or bafilomycin A1, mimicked the cytoprotective effect of 6b. While the treatment with 6b had no effect on the phosphorylation of proapoptotic MAP kinases ERK and JNK, it markedly increased the phosphorylation of the prosurvival kinase Akt in 6-OHDA-treated cells. Akt inhibitor DEBC or RNA interference-mediated Akt silencing reduced the ability of 6b to block 6-OHDA-triggered apoptotic and autophagic responses, thus confirming their dependency on Akt activation. The cytoprotective effect of 6b was also observed in 6-OHDA-treated neuronal PC12 cells, but not in SH-SY5Y or PC12 cells exposed to 1-methyl-4-phenylpyridinium, indicating that the observed neuroprotection was dependent on the cytotoxic stimulus. Because of the ability to prevent 6-OHDA induced apoptotic/autophagic cell death through activation of Akt, the investigated arylpiperazines could be potential candidates for treatment of neurodegenerative diseases.
PB  - Elsevier Ltd.
T2  - Neuropharmacology
T1  - Arylpiperazine-mediated activation of Akt protects SH-SY5Y neuroblastoma cells from 6-hydroxydopamine-induced apoptotic and autophagic death
VL  - 72
DO  - 10.1016/j.neuropharm.2013.04.037
SP  - 224
EP  - 235
ER  - 

@article{
author = "Tovilović-Kovačević, Gordana and Zogović, Nevena and Šoškić, Vukić and Schrattenholz, Andre and Kostić-Rajačić, Slađana and Misirkić Marjanović, Maja and Janjetović, Kristina and Vučićević, Ljubica and Arsikin, Katarina and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2013",
abstract = "We investigated the ability of 19 recently synthesized arylpiperazine compounds to protect human SHSY5Y neuroblastoma cells from the neurotoxin 6-hydroxydopamine (6-OHDA). The compound with the most potent neuroprotective action was N-{3-[2-(4-phenyl-piperazin-1-yl)-ethyl]-phenyl}-picolinamide (6b), which reduced 6-OHDA-induced apoptotic death through stabilization of mitochondrial membrane and subsequent prevention of superoxide production, caspase activation and DNA fragmentation. 6-OHDA-triggered autophagic response was also reduced by 6b, which prevented inactivation of the main autophagy repressor mTOR, upregulation of proautophagic beclin-1, conversion of microtubule-associated protein 1 light chain 3 (LC3)-I to autophagosome-associated LC3-II, as well as intracytoplasmic acidification induced by 6-OHDA. The inhibition of autophagy using LC3b gene silencing or pharmacological autophagy blockers 3-methyladenine or bafilomycin A1, mimicked the cytoprotective effect of 6b. While the treatment with 6b had no effect on the phosphorylation of proapoptotic MAP kinases ERK and JNK, it markedly increased the phosphorylation of the prosurvival kinase Akt in 6-OHDA-treated cells. Akt inhibitor DEBC or RNA interference-mediated Akt silencing reduced the ability of 6b to block 6-OHDA-triggered apoptotic and autophagic responses, thus confirming their dependency on Akt activation. The cytoprotective effect of 6b was also observed in 6-OHDA-treated neuronal PC12 cells, but not in SH-SY5Y or PC12 cells exposed to 1-methyl-4-phenylpyridinium, indicating that the observed neuroprotection was dependent on the cytotoxic stimulus. Because of the ability to prevent 6-OHDA induced apoptotic/autophagic cell death through activation of Akt, the investigated arylpiperazines could be potential candidates for treatment of neurodegenerative diseases.",
publisher = "Elsevier Ltd.",
journal = "Neuropharmacology",
title = "Arylpiperazine-mediated activation of Akt protects SH-SY5Y neuroblastoma cells from 6-hydroxydopamine-induced apoptotic and autophagic death",
volume = "72",
doi = "10.1016/j.neuropharm.2013.04.037",
pages = "224-235"
}

Tovilović-Kovačević, G., Zogović, N., Šoškić, V., Schrattenholz, A., Kostić-Rajačić, S., Misirkić Marjanović, M., Janjetović, K., Vučićević, L., Arsikin, K., Harhaji-Trajković, L.,& Trajković, V.. (2013). Arylpiperazine-mediated activation of Akt protects SH-SY5Y neuroblastoma cells from 6-hydroxydopamine-induced apoptotic and autophagic death. in Neuropharmacology
Elsevier Ltd.., 72, 224-235.
https://doi.org/10.1016/j.neuropharm.2013.04.037

Tovilović-Kovačević G, Zogović N, Šoškić V, Schrattenholz A, Kostić-Rajačić S, Misirkić Marjanović M, Janjetović K, Vučićević L, Arsikin K, Harhaji-Trajković L, Trajković V. Arylpiperazine-mediated activation of Akt protects SH-SY5Y neuroblastoma cells from 6-hydroxydopamine-induced apoptotic and autophagic death. in Neuropharmacology. 2013;72:224-235.
doi:10.1016/j.neuropharm.2013.04.037 .

Tovilović-Kovačević, Gordana, Zogović, Nevena, Šoškić, Vukić, Schrattenholz, Andre, Kostić-Rajačić, Slađana, Misirkić Marjanović, Maja, Janjetović, Kristina, Vučićević, Ljubica, Arsikin, Katarina, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "Arylpiperazine-mediated activation of Akt protects SH-SY5Y neuroblastoma cells from 6-hydroxydopamine-induced apoptotic and autophagic death" in Neuropharmacology, 72 (2013):224-235,
https://doi.org/10.1016/j.neuropharm.2013.04.037 . .

TY  - CONF
AU  - Zogović, Nevena
AU  - Tovilović-Kovačević, Gordana
AU  - Misirkić Marjanović, Maja
AU  - Janjetović, Kristina
AU  - Vučićević, Ljubica
AU  - Trajković, Vladimir
PY  - 2013
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6354
AB  - Introduction: Neural differentiation involves
intracellular signaling-controlled maturation of neural
progenitors into cells with fully developed neuronal
phenotype.
Aim: We explored the interplay between the
adenosine monophosphate-activated protein kinase
(AMPK), extracellular signal-regulated kinase (ERK) and
autophagy in phorbol myristate acetate (PMA)-induced
differentiation of SH-SY5Y human neuroblastoma cells.
Methods: The levels of neuronal marker expression
and acidification of autophagic compartments were
determined by flow cytometry of cells labeled with
appropriate antibodies and acridine orange, respectively.
The activation (phosphorylation) of AMPK and ERK, as
well as the levels of autophagic proteins beclin-1, Atg7,
microtubule-associated protein light chain 3 (LC3) and p62,
were assessed using immunoblot. RNA interference was
used for AMPK and LC3 knockdown.
Results: PMA initiated autophagic response in SH-
SY5Y cells simultaneously with activation of AMPK, a
major intracellular energy sensor, and ERK, a kinase
involved in cell proliferation and differentiation.
Pharmacological inhibition of AMPK or AMPK gene
silencing attenuated differentiation of SH-SY5Y cells, as
well as PMA-induced ERK activation and autophagy. A
selective pharmacological blockade of ERK prevented
PMA-induced neuronal differentiation and autophagy
induction, without affecting AMPK phosphorylation. On the
other hand, the inhibition of autophagy downstream of
AMPK/ERK activation, either by pharmacological agents or
LC3 knockdown, actually promoted the expression of
neuronal markers, thus indicating a role for autophagy in
suppression of PMA-induced differentiation of SH-SY5Y
cells.
Conclusion: PMA-induced differentiation of SH-
SY5Y cells depends on a complex interplay between
AMPK, ERK and autophagy, where AMPK and ERK
promote differentiation independently of autophagy, while
simultaneously inhibiting differentiation process through
autophagy-dependent mechanisms.
AB  - Uvod: Diferencijacija neurona je proces
kontrolisan od strane nekoliko unutarćelijskih signala. U
toku diferencijacije dolazi do potpunog sazrevanja
progenitorskih ćelija u neurone.
Cilj: Cilj ovog rada je bio da se ispita uloga protein
kinaze regulisane adenozin monofosfatom (AMPK), kinaze
regulisane vanćelijskim signalima (ERK) i autofagije tokom
diferencijacije humanih neuroblastoma SH-SY5Y izazvane
forbol miristat acetatom (PMA).
Metode: Ekspresija neuronskih markera i
autofagija su praćene pomoću protočnog citofluorimetra
nakon bojenja ćelija sa odgovarajućim antitelima, odnosno
akridin oranžom. Aktivnosti AMPK, ERK, beklina, Atg7,
proteina lakog lanca 3 povezanog sa mikrotubulima (LC3),
p62, p70S6K i aktina su odreñene pomoću imunoblota.
Utišavanje AMPK i LC3 gena je izvedeno pomoću malih
interferirajućih RNK.
Rezultati: Istovremeno sa započinjanjem
autofagije, PMA je aktivirao AMPK protein, glavni
energetski senzor u ćeliji, i ERK kinazu, zaduženu za
regulaciju diferencijacije i proliferacije. Farmakološka
inhibicija AMPK i utišavanje gena za AMPK usporili su
diferencijaciju SH-SY5Y ćelija, i inhibirali su aktivaciju
ERK kinaze i autofagiju izazvane pomoću PMA. Blokiranje
aktivnosti ERK proteina pomoću specifičnog inhibitora
značajno je usporilo diferencijaciju SH-SY5Y ćelija i
autofagiju izazvane posredstvom PMA, bez uticaja na
aktivnost AMPK. Sa druge strane, inhibicija autofagije
nishodno od AMPK/ERK (farmakološka ili utišavanjem
gena za LC3) je povećala ekspresiju neuronskih markera,
što ukazuje na činjenicu da autofagija izazvana pomoću
PMA najverovatnije suprimira diferencijaciju SH-SY5Y
ćelija.
Zaključak: Diferencijaciju SH-SY5Y ćelija
izazvanu posredstvom PMA pospešuju AMPK i ERK
proteini nezavisno od autofagnog procesa. Istovremeno oba
molekula inhibiraju diferencijaciju preko mehanizama
zavisnih od autofagije.
PB  - Belgrade : Serbian Neuroscience Society
C3  - 6th SNS Serbian Neuroscience Society Congress: book of abstracts; 2013 Nov 14-16; Belgrade, Serbia
T1  - AMPK and ERK regulate phorbol myristate acetate induced differentiation of SH-SY5Y neuroblastoma cells trough autophagy-dependent and –independent mehanisms
T1  - Protein kinaza regulisana adenozin monofosfatom i kinaza regulisana vanćelijskim signalima regulišu diferencijaciju SH-SY5Y ćelija neuroblastoma izazvanu forbol miristat acetatom preko mehanizama zavisnih i nezavisnih od autofagije
SP  - 58
EP  - 58
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6354
ER  - 

@conference{
editor = "Kanazir, Selma, Savić, Danijela, Isaković, Aleksandra",
author = "Zogović, Nevena and Tovilović-Kovačević, Gordana and Misirkić Marjanović, Maja and Janjetović, Kristina and Vučićević, Ljubica and Trajković, Vladimir",
year = "2013",
abstract = "Introduction: Neural differentiation involves
intracellular signaling-controlled maturation of neural
progenitors into cells with fully developed neuronal
phenotype.
Aim: We explored the interplay between the
adenosine monophosphate-activated protein kinase
(AMPK), extracellular signal-regulated kinase (ERK) and
autophagy in phorbol myristate acetate (PMA)-induced
differentiation of SH-SY5Y human neuroblastoma cells.
Methods: The levels of neuronal marker expression
and acidification of autophagic compartments were
determined by flow cytometry of cells labeled with
appropriate antibodies and acridine orange, respectively.
The activation (phosphorylation) of AMPK and ERK, as
well as the levels of autophagic proteins beclin-1, Atg7,
microtubule-associated protein light chain 3 (LC3) and p62,
were assessed using immunoblot. RNA interference was
used for AMPK and LC3 knockdown.
Results: PMA initiated autophagic response in SH-
SY5Y cells simultaneously with activation of AMPK, a
major intracellular energy sensor, and ERK, a kinase
involved in cell proliferation and differentiation.
Pharmacological inhibition of AMPK or AMPK gene
silencing attenuated differentiation of SH-SY5Y cells, as
well as PMA-induced ERK activation and autophagy. A
selective pharmacological blockade of ERK prevented
PMA-induced neuronal differentiation and autophagy
induction, without affecting AMPK phosphorylation. On the
other hand, the inhibition of autophagy downstream of
AMPK/ERK activation, either by pharmacological agents or
LC3 knockdown, actually promoted the expression of
neuronal markers, thus indicating a role for autophagy in
suppression of PMA-induced differentiation of SH-SY5Y
cells.
Conclusion: PMA-induced differentiation of SH-
SY5Y cells depends on a complex interplay between
AMPK, ERK and autophagy, where AMPK and ERK
promote differentiation independently of autophagy, while
simultaneously inhibiting differentiation process through
autophagy-dependent mechanisms., Uvod: Diferencijacija neurona je proces
kontrolisan od strane nekoliko unutarćelijskih signala. U
toku diferencijacije dolazi do potpunog sazrevanja
progenitorskih ćelija u neurone.
Cilj: Cilj ovog rada je bio da se ispita uloga protein
kinaze regulisane adenozin monofosfatom (AMPK), kinaze
regulisane vanćelijskim signalima (ERK) i autofagije tokom
diferencijacije humanih neuroblastoma SH-SY5Y izazvane
forbol miristat acetatom (PMA).
Metode: Ekspresija neuronskih markera i
autofagija su praćene pomoću protočnog citofluorimetra
nakon bojenja ćelija sa odgovarajućim antitelima, odnosno
akridin oranžom. Aktivnosti AMPK, ERK, beklina, Atg7,
proteina lakog lanca 3 povezanog sa mikrotubulima (LC3),
p62, p70S6K i aktina su odreñene pomoću imunoblota.
Utišavanje AMPK i LC3 gena je izvedeno pomoću malih
interferirajućih RNK.
Rezultati: Istovremeno sa započinjanjem
autofagije, PMA je aktivirao AMPK protein, glavni
energetski senzor u ćeliji, i ERK kinazu, zaduženu za
regulaciju diferencijacije i proliferacije. Farmakološka
inhibicija AMPK i utišavanje gena za AMPK usporili su
diferencijaciju SH-SY5Y ćelija, i inhibirali su aktivaciju
ERK kinaze i autofagiju izazvane pomoću PMA. Blokiranje
aktivnosti ERK proteina pomoću specifičnog inhibitora
značajno je usporilo diferencijaciju SH-SY5Y ćelija i
autofagiju izazvane posredstvom PMA, bez uticaja na
aktivnost AMPK. Sa druge strane, inhibicija autofagije
nishodno od AMPK/ERK (farmakološka ili utišavanjem
gena za LC3) je povećala ekspresiju neuronskih markera,
što ukazuje na činjenicu da autofagija izazvana pomoću
PMA najverovatnije suprimira diferencijaciju SH-SY5Y
ćelija.
Zaključak: Diferencijaciju SH-SY5Y ćelija
izazvanu posredstvom PMA pospešuju AMPK i ERK
proteini nezavisno od autofagnog procesa. Istovremeno oba
molekula inhibiraju diferencijaciju preko mehanizama
zavisnih od autofagije.",
publisher = "Belgrade : Serbian Neuroscience Society",
journal = "6th SNS Serbian Neuroscience Society Congress: book of abstracts; 2013 Nov 14-16; Belgrade, Serbia",
title = "AMPK and ERK regulate phorbol myristate acetate induced differentiation of SH-SY5Y neuroblastoma cells trough autophagy-dependent and –independent mehanisms, Protein kinaza regulisana adenozin monofosfatom i kinaza regulisana vanćelijskim signalima regulišu diferencijaciju SH-SY5Y ćelija neuroblastoma izazvanu forbol miristat acetatom preko mehanizama zavisnih i nezavisnih od autofagije",
pages = "58-58",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6354"
}

Kanazir, S., Savić, D., Isaković, A., Zogović, N., Tovilović-Kovačević, G., Misirkić Marjanović, M., Janjetović, K., Vučićević, L.,& Trajković, V.. (2013). AMPK and ERK regulate phorbol myristate acetate induced differentiation of SH-SY5Y neuroblastoma cells trough autophagy-dependent and –independent mehanisms. in 6th SNS Serbian Neuroscience Society Congress: book of abstracts; 2013 Nov 14-16; Belgrade, Serbia
Belgrade : Serbian Neuroscience Society., 58-58.
https://hdl.handle.net/21.15107/rcub_ibiss_6354

Kanazir S, Savić D, Isaković A, Zogović N, Tovilović-Kovačević G, Misirkić Marjanović M, Janjetović K, Vučićević L, Trajković V. AMPK and ERK regulate phorbol myristate acetate induced differentiation of SH-SY5Y neuroblastoma cells trough autophagy-dependent and –independent mehanisms. in 6th SNS Serbian Neuroscience Society Congress: book of abstracts; 2013 Nov 14-16; Belgrade, Serbia. 2013;:58-58.
https://hdl.handle.net/21.15107/rcub_ibiss_6354 .

Kanazir, Selma, Savić, Danijela, Isaković, Aleksandra, Zogović, Nevena, Tovilović-Kovačević, Gordana, Misirkić Marjanović, Maja, Janjetović, Kristina, Vučićević, Ljubica, Trajković, Vladimir, "AMPK and ERK regulate phorbol myristate acetate induced differentiation of SH-SY5Y neuroblastoma cells trough autophagy-dependent and –independent mehanisms" in 6th SNS Serbian Neuroscience Society Congress: book of abstracts; 2013 Nov 14-16; Belgrade, Serbia (2013):58-58,
https://hdl.handle.net/21.15107/rcub_ibiss_6354 .

TY  - THES
AU  - Vučićević, Ljubica
PY  - 2013
UR  - http://eteze.bg.ac.rs/application/showtheses?thesesId=786
UR  - https://fedorabg.bg.ac.rs/fedora/get/o:7104/bdef:Content/download
UR  - http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=1024639666
UR  - http://nardus.mpn.gov.rs/123456789/2112
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2431
AB  - U ovoj doktorskoj disertaciji ispitivan je uticaj inhibicije intracelularnog energetskog senzora protein kinaze aktivirane adenozin-monofosfatom (AMPK) na indukciju apoptoze i autofagije u tumorskim ćelija. Farmakološki inhibitor AMPK dorzomorfin indukovao je G2/M blokadu ćelijskog ciklusa, praćen apoptozom koju karakteriše aktivacija kaspaza, eksternalizacija fosfatidilserina i fragmentacija DNK u U251 humanim i C6 pacovskim ćelijama glioma, dok na vijabilitet primarnih pacovskih astrocita i ćelija mišjeg melanoma B16 nije imao uticaja. Mehanizam indukcije apoptoze dorzomorfinom bio je posredovan stimulacijom produkcije reaktivnih vrsta kiseonika i inhibicijom ekspresije antiapoptotskog Bcl-2 proteina. Dorzomorfin je inhibirao fosforilaciju i enzimatsku aktivnost AMPK, što je za posledicu imalo smanjenje fosforilacije njenog supstrata acetil-CoA karboksilaze. Aktivatori AMPK metformin i AICAR delimično su neutralisali blokadu ćelijskog ciklusa, oksidativni stres i apoptozu indukovanu dorzomorfinom. Mala interferirajuća RNK (siRNK) koja sprečava ekspresiju humanog AMPK enzima je poput dorzomorfina zaustavila proliferaciju ćelija u G2/M fazi ćelijskog ciklusa, ali nije izazvala oksidativni stres i apoptozu u U251 ćelijama. Dakle, inhibicija AMPK je neophodna, ali ne i dovoljna za indukciju apotpoze dorzomorfinom u ćelijama glioma.
U ovoj studiji takođe je pokazano da dorzomorfin indukuje autofagiju u ćelijama kancera. Indukcija autofagije u U251 ćelijama detektovana je fluorescentnim bojenjem unutarćelijskih kiselih vezikula akridin oranžom, indukcijom beklina-1, degradacijom p62 proteina i konverzijom LC3-I u formu asociranu sa autofagozomima LC3-II u odsustvu i
prisustvu proteolitičkih inhibitora. Prisustvo autofagozomima sličnih vezikula potvrđeno je transmisionom elektronskom mikroskopijom. Inhibicija AMPK i Raptora indukovana dorzomorfinom u U251 ćelijama bila je paradoksalno asocirana sa smanjenjem fosforilacije AMPK/Raptorom inhibiranog glavnog represora autofagije mTOR i njegovog supstrata p70S6K. Fosforilacija mTOR aktivatora Akt i PI3K-aktivirane Src kinaze bile su takođe inhibirane u ćelijama tretiranim dorzomorfinom. Poništavanje ekspresije AMPK sa siRNA nije redukovalo aktivnost Akt/mTOR/p70S6K signalnog puta, a AMPK aktivatori metformin i AICAR nisu uspeli da blokiraju autofagiju indukovanu dorzomorfinom. Inhibitori autofagije bafilomicin i hlorokin značajno su povećali citotoksičnost dorzomorfina prema U251 ćelijama, što je pokazano povećanjem oslobađanja laktat dehidrogenaze, fragmentacije DNK i aktivacije kaspaze-3. Sličan efekat dorzomorfin je imao i prema ćelijama pacovskog glioma C6, mišjeg fibrosarkoma L929 i mišjeg melanoma B16. Pošto je ranije pokazano da dorzomorfin suprimira AMPK-zavisnu autofagiju u različitim ćelijskim tipovima, rezultati ove studije sugerišu da efekat dorzomorfina na autofagiju zavisi od doze, konteksta i/ili vrste ćelija. Imajući u vidu da dorzomorfin indukuje autofagiju u ćelijama kancera nezavisno od inhibicije AMPK, inhibicijom Akt/mTOR signalnog puta, rezultati ove studije ukazuju na neophodnost opreza pri interpretaciji rezultata eksperimenata u kojima se dorzomorfin koristi kao inhibitor AMPK, ali takođe sugerišu da bi dorzomorfin, sam ili u kombinaciji sa inhibitorima autofagije, mogao biti potencijalni kandidat za terapiju tumora.
AB  - In this doctoral dissertation the effect of intracellular energy sensor AMP-activated protein kinase (AMPK) inhibition on induction of apoptosis and autophagy in tumor cells was investigated. Pharmacological AMPK inhibitor dorsomorphin caused G2/M cell cycle block, accompanied by apoptotic cell death characterized by caspase activation, phosphatidylserine exposure and DNA fragmentation in U251 human and C6 rat glioma cells, while it had no effect on viability of primary rat astrocytes and B16 mouse melanoma cells. The mechanisms underlying the pro-apoptotic action of dorsomorphin involved induction of oxidative stress and down-regulation of antiapoptotic molecule Bcl-2. Dorsomorphin diminished AMPK phosphorylation and enzymatic activity, resulting in reduced phosphorylation of its target acetyl CoA carboxylase. AMPK activators metformin and AICAR partly prevented the cell cycle block, oxidative stress and apoptosis induced by dorsomorphin. The small interfering RNA (siRNA) targeting of human AMPK mimicked dorsomorphin-induced G2/M cell cycle arrest, but failed to induce oxidative stress and apoptosis in U251 glioma cells. Therefore, AMPK inhibition is required, but not sufficient for dorsomorphin-mediated apoptotic death of glioma cells.
In this study, it was also reported that dorsomorphin can induce autophagy in cancer cells. The induction of autophagy in U251 human glioma cell line was demonstrated by acridine orange staining of intracellular acidic vesicles, Beclin 1 induction, p62 decrease and conversion of LC3-I to autophagosome-associated LC3-II in the absence and presence of proteolysis inhibitors. The presence of autophagosome like vesicles was confirmed by transmission electron microscopy. Dorsomorphin-mediated inhibition of AMPK and Raptor in U251 cells was associated with paradoxical decrease in phosphorylation of
AMPK/Raptor-repressed mTOR, a major negative regulator of autophagy, and its downstream target p70S6K. The phosphorylation of mTOR activator Akt and PI3K-activating kinase Src was also impaired in dorsomorphin-treated cells. The siRNA-mediated AMPK silencing did not reduce the activity of the Akt/mTOR/p70S6K pathway and AMPK activators metformin and AICAR failed to block dorsomorphin-induced autophagy. Autophagy inhibitors bafilomycin and chloroquine significantly increased the cytotoxicity of dorsomorphin towards U251 cells, as confirmed by the increase in lactate dehydrogenase release, DNA fragmentation and caspase-3 activation. Similar effects of dorsomorphin were also observed in C6 rat glioma, L929 mouse fibrosarcoma and B16 mouse melanoma cell lines. Since dorsomorphin has previously been reported to suppress AMPK dependent autophagy in different cell types, results in this study suggest that the effects of dorsomorphin on autophagy might be dose-, cell type- and/or context-dependent. By demonstrating the ability of dorsomorphin to induce autophagic response in cancer cells via AMPK inhibition-independent downregulation of Akt/mTOR pathway, results warrant caution when using dorsomorphin to inhibit AMPK-dependent cellular responses, but also suggest that dorsomorphin, alone or in combination with autophagy inhibitors, could be potential candidate for anticancer therapy.
PB  - Belgrade: University of Belgrade, Faculty of Biology
T2  - University of Belgrade, Faculty of Biology
T1  - Uloga inhibicije protein-kinaze aktivirane adenozin-monofosfatom u indukciji apoptoze i autofagije u tumorskim ćelijskim linijama
T1  - The role of adenosine monophosphate-activated protein kinase inhibition in apoptosis and autophagy induction in tumor cell lines
SP  - 1
EP  - 90
UR  - https://hdl.handle.net/21.15107/rcub_nardus_2112
ER  - 

@phdthesis{
author = "Vučićević, Ljubica",
year = "2013",
abstract = "U ovoj doktorskoj disertaciji ispitivan je uticaj inhibicije intracelularnog energetskog senzora protein kinaze aktivirane adenozin-monofosfatom (AMPK) na indukciju apoptoze i autofagije u tumorskim ćelija. Farmakološki inhibitor AMPK dorzomorfin indukovao je G2/M blokadu ćelijskog ciklusa, praćen apoptozom koju karakteriše aktivacija kaspaza, eksternalizacija fosfatidilserina i fragmentacija DNK u U251 humanim i C6 pacovskim ćelijama glioma, dok na vijabilitet primarnih pacovskih astrocita i ćelija mišjeg melanoma B16 nije imao uticaja. Mehanizam indukcije apoptoze dorzomorfinom bio je posredovan stimulacijom produkcije reaktivnih vrsta kiseonika i inhibicijom ekspresije antiapoptotskog Bcl-2 proteina. Dorzomorfin je inhibirao fosforilaciju i enzimatsku aktivnost AMPK, što je za posledicu imalo smanjenje fosforilacije njenog supstrata acetil-CoA karboksilaze. Aktivatori AMPK metformin i AICAR delimično su neutralisali blokadu ćelijskog ciklusa, oksidativni stres i apoptozu indukovanu dorzomorfinom. Mala interferirajuća RNK (siRNK) koja sprečava ekspresiju humanog AMPK enzima je poput dorzomorfina zaustavila proliferaciju ćelija u G2/M fazi ćelijskog ciklusa, ali nije izazvala oksidativni stres i apoptozu u U251 ćelijama. Dakle, inhibicija AMPK je neophodna, ali ne i dovoljna za indukciju apotpoze dorzomorfinom u ćelijama glioma.
U ovoj studiji takođe je pokazano da dorzomorfin indukuje autofagiju u ćelijama kancera. Indukcija autofagije u U251 ćelijama detektovana je fluorescentnim bojenjem unutarćelijskih kiselih vezikula akridin oranžom, indukcijom beklina-1, degradacijom p62 proteina i konverzijom LC3-I u formu asociranu sa autofagozomima LC3-II u odsustvu i
prisustvu proteolitičkih inhibitora. Prisustvo autofagozomima sličnih vezikula potvrđeno je transmisionom elektronskom mikroskopijom. Inhibicija AMPK i Raptora indukovana dorzomorfinom u U251 ćelijama bila je paradoksalno asocirana sa smanjenjem fosforilacije AMPK/Raptorom inhibiranog glavnog represora autofagije mTOR i njegovog supstrata p70S6K. Fosforilacija mTOR aktivatora Akt i PI3K-aktivirane Src kinaze bile su takođe inhibirane u ćelijama tretiranim dorzomorfinom. Poništavanje ekspresije AMPK sa siRNA nije redukovalo aktivnost Akt/mTOR/p70S6K signalnog puta, a AMPK aktivatori metformin i AICAR nisu uspeli da blokiraju autofagiju indukovanu dorzomorfinom. Inhibitori autofagije bafilomicin i hlorokin značajno su povećali citotoksičnost dorzomorfina prema U251 ćelijama, što je pokazano povećanjem oslobađanja laktat dehidrogenaze, fragmentacije DNK i aktivacije kaspaze-3. Sličan efekat dorzomorfin je imao i prema ćelijama pacovskog glioma C6, mišjeg fibrosarkoma L929 i mišjeg melanoma B16. Pošto je ranije pokazano da dorzomorfin suprimira AMPK-zavisnu autofagiju u različitim ćelijskim tipovima, rezultati ove studije sugerišu da efekat dorzomorfina na autofagiju zavisi od doze, konteksta i/ili vrste ćelija. Imajući u vidu da dorzomorfin indukuje autofagiju u ćelijama kancera nezavisno od inhibicije AMPK, inhibicijom Akt/mTOR signalnog puta, rezultati ove studije ukazuju na neophodnost opreza pri interpretaciji rezultata eksperimenata u kojima se dorzomorfin koristi kao inhibitor AMPK, ali takođe sugerišu da bi dorzomorfin, sam ili u kombinaciji sa inhibitorima autofagije, mogao biti potencijalni kandidat za terapiju tumora., In this doctoral dissertation the effect of intracellular energy sensor AMP-activated protein kinase (AMPK) inhibition on induction of apoptosis and autophagy in tumor cells was investigated. Pharmacological AMPK inhibitor dorsomorphin caused G2/M cell cycle block, accompanied by apoptotic cell death characterized by caspase activation, phosphatidylserine exposure and DNA fragmentation in U251 human and C6 rat glioma cells, while it had no effect on viability of primary rat astrocytes and B16 mouse melanoma cells. The mechanisms underlying the pro-apoptotic action of dorsomorphin involved induction of oxidative stress and down-regulation of antiapoptotic molecule Bcl-2. Dorsomorphin diminished AMPK phosphorylation and enzymatic activity, resulting in reduced phosphorylation of its target acetyl CoA carboxylase. AMPK activators metformin and AICAR partly prevented the cell cycle block, oxidative stress and apoptosis induced by dorsomorphin. The small interfering RNA (siRNA) targeting of human AMPK mimicked dorsomorphin-induced G2/M cell cycle arrest, but failed to induce oxidative stress and apoptosis in U251 glioma cells. Therefore, AMPK inhibition is required, but not sufficient for dorsomorphin-mediated apoptotic death of glioma cells.
In this study, it was also reported that dorsomorphin can induce autophagy in cancer cells. The induction of autophagy in U251 human glioma cell line was demonstrated by acridine orange staining of intracellular acidic vesicles, Beclin 1 induction, p62 decrease and conversion of LC3-I to autophagosome-associated LC3-II in the absence and presence of proteolysis inhibitors. The presence of autophagosome like vesicles was confirmed by transmission electron microscopy. Dorsomorphin-mediated inhibition of AMPK and Raptor in U251 cells was associated with paradoxical decrease in phosphorylation of
AMPK/Raptor-repressed mTOR, a major negative regulator of autophagy, and its downstream target p70S6K. The phosphorylation of mTOR activator Akt and PI3K-activating kinase Src was also impaired in dorsomorphin-treated cells. The siRNA-mediated AMPK silencing did not reduce the activity of the Akt/mTOR/p70S6K pathway and AMPK activators metformin and AICAR failed to block dorsomorphin-induced autophagy. Autophagy inhibitors bafilomycin and chloroquine significantly increased the cytotoxicity of dorsomorphin towards U251 cells, as confirmed by the increase in lactate dehydrogenase release, DNA fragmentation and caspase-3 activation. Similar effects of dorsomorphin were also observed in C6 rat glioma, L929 mouse fibrosarcoma and B16 mouse melanoma cell lines. Since dorsomorphin has previously been reported to suppress AMPK dependent autophagy in different cell types, results in this study suggest that the effects of dorsomorphin on autophagy might be dose-, cell type- and/or context-dependent. By demonstrating the ability of dorsomorphin to induce autophagic response in cancer cells via AMPK inhibition-independent downregulation of Akt/mTOR pathway, results warrant caution when using dorsomorphin to inhibit AMPK-dependent cellular responses, but also suggest that dorsomorphin, alone or in combination with autophagy inhibitors, could be potential candidate for anticancer therapy.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology",
journal = "University of Belgrade, Faculty of Biology",
title = "Uloga inhibicije protein-kinaze aktivirane adenozin-monofosfatom u indukciji apoptoze i autofagije u tumorskim ćelijskim linijama, The role of adenosine monophosphate-activated protein kinase inhibition in apoptosis and autophagy induction in tumor cell lines",
pages = "1-90",
url = "https://hdl.handle.net/21.15107/rcub_nardus_2112"
}

Vučićević, L.. (2013). Uloga inhibicije protein-kinaze aktivirane adenozin-monofosfatom u indukciji apoptoze i autofagije u tumorskim ćelijskim linijama. in University of Belgrade, Faculty of Biology
Belgrade: University of Belgrade, Faculty of Biology., 1-90.
https://hdl.handle.net/21.15107/rcub_nardus_2112

Vučićević L. Uloga inhibicije protein-kinaze aktivirane adenozin-monofosfatom u indukciji apoptoze i autofagije u tumorskim ćelijskim linijama. in University of Belgrade, Faculty of Biology. 2013;:1-90.
https://hdl.handle.net/21.15107/rcub_nardus_2112 .

Vučićević, Ljubica, "Uloga inhibicije protein-kinaze aktivirane adenozin-monofosfatom u indukciji apoptoze i autofagije u tumorskim ćelijskim linijama" in University of Belgrade, Faculty of Biology (2013):1-90,
https://hdl.handle.net/21.15107/rcub_nardus_2112 .