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  - CONF
AU  - Ristić, Biljana
AU  - Krunić, Matija
AU  - Paunović, Verica
AU  - Bošnjak, Mihajlo
AU  - Tovilović-Kovačević, Gordana
AU  - Zogović, Nevena
AU  - Mirčić, Aleksandar
AU  - Vuković, Irena
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/123456789/6284
AB  - Introduction: We examined the molecular mechanisms of graphene quantum dot (GQD)- mediated
protection of SH-SY5Y human neuroblastoma cells from oxidative/nitrosative stress induced by iron-nitrosyl complex sodium nitroprusside (SNP).
Methods: GQD was produced by electrochemical oxidation of graphite and characterized by AFM, UVVIS and FTIR spectroscopy. The antioxidant activity of GQD in cell-free conditions was assessed by DPPH,
NBT and EPR analysis. The neuroprotective potential of GQD was determined by cell viability assays MTT,
CV. Flow cytometry was used to assess markers of apoptosis and GQD scavenging of intracellular
ROS/RNS as well. Cellular internalization of GQD was determined using TEM.
Results: GQD prevented SNP-induced apoptosis, caspase activation and mitochondrial depolarization
in neuroblastoma cells. Although GQD diminished the NO levelsin SNP-treated cells, NO scavengers displayed only a slight protection. GQD significantly protected SH-SY5Y cells from neurotoxicity of lightexhausted SNP, incapable of producing NO, implying that protective mechanism is independent of
NO-scavenging. GQD reduced SNP-triggered increase in intracellular levels of ROS, particularly •OH, O2•−
in cells and cell-free condition. Nonselective antioxidants, •OH scavengers and iron chelators, mimicked
GQD cytoprotection, indicating that GQD protect cells by neutralizing •OH generated in the Fenton reaction. Cellular GQD internalization wasrequired for optimal protection since the removal of extracellular GQD by extensive washing partly diminished their protective effect, suggesting that GQD exerted
neuroprotective effect intra- and extracellularly.
Conclusion: By demonstrating that GQD protect neuroblastoma cells from SNP-induced apoptosis by
•OH/NO scavenging, our results suggest that GQD could be valuable candidates for treatment of neurodegenerative diseases associated with oxidative/nitrosative stress.
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  - Graphen quantum dots protect SH-SY5Y neuronal cells from SNP-indced apoptotic death
SP  - 27
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6284
ER  - 

@conference{
author = "Ristić, Biljana and Krunić, Matija and Paunović, Verica and Bošnjak, Mihajlo and Tovilović-Kovačević, Gordana and Zogović, Nevena and Mirčić, Aleksandar and Vuković, Irena and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2023",
abstract = "Introduction: We examined the molecular mechanisms of graphene quantum dot (GQD)- mediated
protection of SH-SY5Y human neuroblastoma cells from oxidative/nitrosative stress induced by iron-nitrosyl complex sodium nitroprusside (SNP).
Methods: GQD was produced by electrochemical oxidation of graphite and characterized by AFM, UVVIS and FTIR spectroscopy. The antioxidant activity of GQD in cell-free conditions was assessed by DPPH,
NBT and EPR analysis. The neuroprotective potential of GQD was determined by cell viability assays MTT,
CV. Flow cytometry was used to assess markers of apoptosis and GQD scavenging of intracellular
ROS/RNS as well. Cellular internalization of GQD was determined using TEM.
Results: GQD prevented SNP-induced apoptosis, caspase activation and mitochondrial depolarization
in neuroblastoma cells. Although GQD diminished the NO levelsin SNP-treated cells, NO scavengers displayed only a slight protection. GQD significantly protected SH-SY5Y cells from neurotoxicity of lightexhausted SNP, incapable of producing NO, implying that protective mechanism is independent of
NO-scavenging. GQD reduced SNP-triggered increase in intracellular levels of ROS, particularly •OH, O2•−
in cells and cell-free condition. Nonselective antioxidants, •OH scavengers and iron chelators, mimicked
GQD cytoprotection, indicating that GQD protect cells by neutralizing •OH generated in the Fenton reaction. Cellular GQD internalization wasrequired for optimal protection since the removal of extracellular GQD by extensive washing partly diminished their protective effect, suggesting that GQD exerted
neuroprotective effect intra- and extracellularly.
Conclusion: By demonstrating that GQD protect neuroblastoma cells from SNP-induced apoptosis by
•OH/NO scavenging, our results suggest that GQD could be valuable candidates for treatment of neurodegenerative diseases associated with oxidative/nitrosative stress.",
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 = "Graphen quantum dots protect SH-SY5Y neuronal cells from SNP-indced apoptotic death",
pages = "27",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6284"
}

Ristić, B., Krunić, M., Paunović, V., Bošnjak, M., Tovilović-Kovačević, G., Zogović, N., Mirčić, A., Vuković, I., Harhaji-Trajković, L.,& Trajković, V.. (2023). Graphen quantum dots protect SH-SY5Y neuronal cells from SNP-indced apoptotic death. 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., 27.
https://hdl.handle.net/21.15107/rcub_ibiss_6284

Ristić B, Krunić M, Paunović V, Bošnjak M, Tovilović-Kovačević G, Zogović N, Mirčić A, Vuković I, Harhaji-Trajković L, Trajković V. Graphen quantum dots protect SH-SY5Y neuronal cells from SNP-indced apoptotic death. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. 2023;:27.
https://hdl.handle.net/21.15107/rcub_ibiss_6284 .

Ristić, Biljana, Krunić, Matija, Paunović, Verica, Bošnjak, Mihajlo, Tovilović-Kovačević, Gordana, Zogović, Nevena, Mirčić, Aleksandar, Vuković, Irena, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "Graphen quantum dots protect SH-SY5Y neuronal cells from SNP-indced apoptotic death" in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia (2023):27,
https://hdl.handle.net/21.15107/rcub_ibiss_6284 .

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  - JOUR
AU  - Saksida, Tamara
AU  - Paunović, Verica
AU  - Koprivica, Ivan
AU  - Mićanović, Dragica
AU  - Jevtić, Bojan
AU  - Jonić, Natalija
AU  - Stojanović, Ivana D.
AU  - Pejnović, Nada
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5557
AB  - Recent data indicate the link between the number and function of T regulatory cells (Treg)
in the gut immune tissue and initiation and development of autoimmunity associated with type
1 diabetes (T1D). Since type 3 innate lymphoid cells (ILC3) in the small intestine are essential for
maintaining FoxP3+ Treg and there are no data about the possible role of ILC3 in T1D pathogenesis,
the aim of this study was to explore ILC3-Treg link during the development of T1D. Mature diabetic
NOD mice had lower frequencies of IL-2-producing ILC3 and Treg in small intestine lamina propria
(SILP) compared to prediabetic NOD mice. Similarly, in multiple low doses of streptozotocin (MLDS)-
induced T1D in C57BL/6 mice, hyperglycemic mice exhibited lower numbers of ILC3, IL-2+ ILC3
and Treg in SILP compared to healthy controls. To boost T1D severity, mice were treated with
broad-spectrum antibiotics (ABX) for 14 days prior to T1D induction by MLDS. The higher incidence
of T1D in ABX-treated mice was associated with significantly lower frequencies of IL-2+ ILC3 and
FoxP3+ Treg in SILP compared with mice without ABX treatment. The obtained findings show that
the lower proportions of IL-2-expressing ILC3 and FoxP3+ Treg in SILP coincided with diabetes
progression and severity.
PB  - MDPI
PB  - Basel: MDPI
T2  - Molecules
T1  - Development of Type 1 Diabetes in Mice Is Associated with a Decrease in IL-2-Producing ILC3 and FoxP3+ Treg in the Small Intestine
IS  - 8
VL  - 28
DO  - 10.3390/molecules28083366
SP  - 3366
ER  - 

@article{
author = "Saksida, Tamara and Paunović, Verica and Koprivica, Ivan and Mićanović, Dragica and Jevtić, Bojan and Jonić, Natalija and Stojanović, Ivana D. and Pejnović, Nada",
year = "2023",
abstract = "Recent data indicate the link between the number and function of T regulatory cells (Treg)
in the gut immune tissue and initiation and development of autoimmunity associated with type
1 diabetes (T1D). Since type 3 innate lymphoid cells (ILC3) in the small intestine are essential for
maintaining FoxP3+ Treg and there are no data about the possible role of ILC3 in T1D pathogenesis,
the aim of this study was to explore ILC3-Treg link during the development of T1D. Mature diabetic
NOD mice had lower frequencies of IL-2-producing ILC3 and Treg in small intestine lamina propria
(SILP) compared to prediabetic NOD mice. Similarly, in multiple low doses of streptozotocin (MLDS)-
induced T1D in C57BL/6 mice, hyperglycemic mice exhibited lower numbers of ILC3, IL-2+ ILC3
and Treg in SILP compared to healthy controls. To boost T1D severity, mice were treated with
broad-spectrum antibiotics (ABX) for 14 days prior to T1D induction by MLDS. The higher incidence
of T1D in ABX-treated mice was associated with significantly lower frequencies of IL-2+ ILC3 and
FoxP3+ Treg in SILP compared with mice without ABX treatment. The obtained findings show that
the lower proportions of IL-2-expressing ILC3 and FoxP3+ Treg in SILP coincided with diabetes
progression and severity.",
publisher = "MDPI, Basel: MDPI",
journal = "Molecules",
title = "Development of Type 1 Diabetes in Mice Is Associated with a Decrease in IL-2-Producing ILC3 and FoxP3+ Treg in the Small Intestine",
number = "8",
volume = "28",
doi = "10.3390/molecules28083366",
pages = "3366"
}

Saksida, T., Paunović, V., Koprivica, I., Mićanović, D., Jevtić, B., Jonić, N., Stojanović, I. D.,& Pejnović, N.. (2023). Development of Type 1 Diabetes in Mice Is Associated with a Decrease in IL-2-Producing ILC3 and FoxP3+ Treg in the Small Intestine. in Molecules
MDPI., 28(8), 3366.
https://doi.org/10.3390/molecules28083366

Saksida T, Paunović V, Koprivica I, Mićanović D, Jevtić B, Jonić N, Stojanović ID, Pejnović N. Development of Type 1 Diabetes in Mice Is Associated with a Decrease in IL-2-Producing ILC3 and FoxP3+ Treg in the Small Intestine. in Molecules. 2023;28(8):3366.
doi:10.3390/molecules28083366 .

Saksida, Tamara, Paunović, Verica, Koprivica, Ivan, Mićanović, Dragica, Jevtić, Bojan, Jonić, Natalija, Stojanović, Ivana D., Pejnović, Nada, "Development of Type 1 Diabetes in Mice Is Associated with a Decrease in IL-2-Producing ILC3 and FoxP3+ Treg in the Small Intestine" in Molecules, 28, no. 8 (2023):3366,
https://doi.org/10.3390/molecules28083366 . .

TY  - DATA
AU  - Đorđević, Marija
AU  - Stepper, Peter
AU  - Feuerstein-Akgoz, Clarissa
AU  - Gerhauser, Clarissa
AU  - Paunović, Verica
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Jurkowska, Renata
AU  - Jurkowski, Tomasz
AU  - Arambašić Jovanović, Jelena
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5669
AB  - Figure 1. UCSC genome browser views of murine Arx gene’s DNA methylation sites in the pancreas and other tissues. If compared with other tissues, Arx gene in pancreatic tissue (young and old) exhibits low DNA methylation. CpG island was shown as a green box. Table 1. Targeted sequences for sgRNAs. Table 2. Primers used for RT-qPCR. Table 3. Primers used for HRM. Table 4. Touchdown PCR program for amplification of bisulfite converted DNA, starting at 55 °C. Table 5. Primers for NGS library preparation. Table 6. Antibodies used for Immunoblot analysis (IBA) and Immunocytochemistry (ICC). Table 7. Primers for PCR reaction after ChIP.
PB  - Frontiers Media S.A.
T2  - Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
T1  - Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.
VL  - 14
SP  - 1134478
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5669
ER  - 

@misc{
author = "Đorđević, Marija and Stepper, Peter and Feuerstein-Akgoz, Clarissa and Gerhauser, Clarissa and Paunović, Verica and Tolić, Anja and Rajić, Jovana and Dinić, Svetlana and Uskoković, Aleksandra and Grdović, Nevena and Mihailović, Mirjana and Jurkowska, Renata and Jurkowski, Tomasz and Arambašić Jovanović, Jelena and Vidaković, Melita",
year = "2023",
abstract = "Figure 1. UCSC genome browser views of murine Arx gene’s DNA methylation sites in the pancreas and other tissues. If compared with other tissues, Arx gene in pancreatic tissue (young and old) exhibits low DNA methylation. CpG island was shown as a green box. Table 1. Targeted sequences for sgRNAs. Table 2. Primers used for RT-qPCR. Table 3. Primers used for HRM. Table 4. Touchdown PCR program for amplification of bisulfite converted DNA, starting at 55 °C. Table 5. Primers for NGS library preparation. Table 6. Antibodies used for Immunoblot analysis (IBA) and Immunocytochemistry (ICC). Table 7. Primers for PCR reaction after ChIP.",
publisher = "Frontiers Media S.A.",
journal = "Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms",
title = "Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.",
volume = "14",
pages = "1134478",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5669"
}

Đorđević, M., Stepper, P., Feuerstein-Akgoz, C., Gerhauser, C., Paunović, V., Tolić, A., Rajić, J., Dinić, S., Uskoković, A., Grdović, N., Mihailović, M., Jurkowska, R., Jurkowski, T., Arambašić Jovanović, J.,& Vidaković, M.. (2023). Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
Frontiers Media S.A.., 14, 1134478.
https://hdl.handle.net/21.15107/rcub_ibiss_5669

Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska R, Jurkowski T, Arambašić Jovanović J, Vidaković M. Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478.. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms. 2023;14:1134478.
https://hdl.handle.net/21.15107/rcub_ibiss_5669 .

Đorđević, Marija, Stepper, Peter, Feuerstein-Akgoz, Clarissa, Gerhauser, Clarissa, Paunović, Verica, Tolić, Anja, Rajić, Jovana, Dinić, Svetlana, Uskoković, Aleksandra, Grdović, Nevena, Mihailović, Mirjana, Jurkowska, Renata, Jurkowski, Tomasz, Arambašić Jovanović, Jelena, Vidaković, Melita, "Supplementary Material for Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska RZ, Jurkowski TP, Jovanović JA, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. Front Endocrinol (Lausanne). 2023;14:1134478." in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms, 14 (2023):1134478,
https://hdl.handle.net/21.15107/rcub_ibiss_5669 .

TY  - JOUR
AU  - Đorđević, Marija
AU  - Stepper, Peter
AU  - Feuerstein-Akgoz, Clarissa
AU  - Gerhauser, Clarissa
AU  - Paunović, Verica
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Jurkowska, Renata
AU  - Jurkowski, Tomasz
AU  - Arambašić Jovanović, Jelena
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5507
AB  - Introduction: Beta cell dysfunction by loss of beta cell identity, dedifferentiation, and the presence of polyhormonal cells are main characteristics of diabetes. The straightforward strategy for curing diabetes implies reestablishment of pancreatic beta cell function by beta cell replacement therapy. Aristaless-related homeobox (Arx) gene encodes protein which plays an important role in the development of pancreatic alpha cells and is a main target for changing alpha cell identity.

Results: In this study we used CRISPR/dCas9-based epigenetic tools for targeted hypermethylation of Arx gene promoter and its subsequent suppression in mouse pancreatic αTC1-6 cell line. Bisulfite sequencing and methylation profiling revealed that the dCas9-Dnmt3a3L-KRAB single chain fusion constructs (EpiCRISPR) was the most efficient. Epigenetic silencing of Arx expression was accompanied by an increase in transcription of the insulin gene (Ins2) mRNA on 5th and 7th post-transfection day, quantified by both RT-qPCR and RNA-seq. Insulin production and secretion was determined by immunocytochemistry and ELISA assay, respectively. Eventually, we were able to induce switch of approximately 1% of transiently transfected cells which were able to produce 35% more insulin than Mock transfected alpha cells.

Conclusion: In conclusion, we successfully triggered a direct, transient switch of pancreatic alpha to insulin-producing cells opening a future research on promising therapeutic avenue for diabetes management.
PB  - Frontiers Media S.A.
T2  - Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
T1  - EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells
VL  - 14
DO  - 10.3389/fendo.2023.1134478
SP  - 1134478
ER  - 

@article{
author = "Đorđević, Marija and Stepper, Peter and Feuerstein-Akgoz, Clarissa and Gerhauser, Clarissa and Paunović, Verica and Tolić, Anja and Rajić, Jovana and Dinić, Svetlana and Uskoković, Aleksandra and Grdović, Nevena and Mihailović, Mirjana and Jurkowska, Renata and Jurkowski, Tomasz and Arambašić Jovanović, Jelena and Vidaković, Melita",
year = "2023",
abstract = "Introduction: Beta cell dysfunction by loss of beta cell identity, dedifferentiation, and the presence of polyhormonal cells are main characteristics of diabetes. The straightforward strategy for curing diabetes implies reestablishment of pancreatic beta cell function by beta cell replacement therapy. Aristaless-related homeobox (Arx) gene encodes protein which plays an important role in the development of pancreatic alpha cells and is a main target for changing alpha cell identity.

Results: In this study we used CRISPR/dCas9-based epigenetic tools for targeted hypermethylation of Arx gene promoter and its subsequent suppression in mouse pancreatic αTC1-6 cell line. Bisulfite sequencing and methylation profiling revealed that the dCas9-Dnmt3a3L-KRAB single chain fusion constructs (EpiCRISPR) was the most efficient. Epigenetic silencing of Arx expression was accompanied by an increase in transcription of the insulin gene (Ins2) mRNA on 5th and 7th post-transfection day, quantified by both RT-qPCR and RNA-seq. Insulin production and secretion was determined by immunocytochemistry and ELISA assay, respectively. Eventually, we were able to induce switch of approximately 1% of transiently transfected cells which were able to produce 35% more insulin than Mock transfected alpha cells.

Conclusion: In conclusion, we successfully triggered a direct, transient switch of pancreatic alpha to insulin-producing cells opening a future research on promising therapeutic avenue for diabetes management.",
publisher = "Frontiers Media S.A.",
journal = "Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms",
title = "EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells",
volume = "14",
doi = "10.3389/fendo.2023.1134478",
pages = "1134478"
}

Đorđević, M., Stepper, P., Feuerstein-Akgoz, C., Gerhauser, C., Paunović, V., Tolić, A., Rajić, J., Dinić, S., Uskoković, A., Grdović, N., Mihailović, M., Jurkowska, R., Jurkowski, T., Arambašić Jovanović, J.,& Vidaković, M.. (2023). EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms
Frontiers Media S.A.., 14, 1134478.
https://doi.org/10.3389/fendo.2023.1134478

Đorđević M, Stepper P, Feuerstein-Akgoz C, Gerhauser C, Paunović V, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Jurkowska R, Jurkowski T, Arambašić Jovanović J, Vidaković M. EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells. in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms. 2023;14:1134478.
doi:10.3389/fendo.2023.1134478 .

Đorđević, Marija, Stepper, Peter, Feuerstein-Akgoz, Clarissa, Gerhauser, Clarissa, Paunović, Verica, Tolić, Anja, Rajić, Jovana, Dinić, Svetlana, Uskoković, Aleksandra, Grdović, Nevena, Mihailović, Mirjana, Jurkowska, Renata, Jurkowski, Tomasz, Arambašić Jovanović, Jelena, Vidaković, Melita, "EpiCRISPR targeted methylation of Arx gene initiates transient switch of mouse pancreatic alpha to insulin-producing cells" in Frontiers in Endocrinology, Section - Diabetes: Molecular Mechanisms, 14 (2023):1134478,
https://doi.org/10.3389/fendo.2023.1134478 . .

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  - 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  - JOUR
AU  - Đorđević, Marija
AU  - Paunović, Verica
AU  - Jovanović Tucović, Maja
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Marković, Ivanka
AU  - Arambašić Jovanović, Jelena
AU  - Vidaković, Melita
PY  - 2022
UR  - https://www.mdpi.com/2076-3417/12/15/7938
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5102
AB  - An efficient transfection is a crucial step for the introduction of epigenetic modification in host cells, and there is a need for an optimized transfection process for individual model systems separately. Mouse pancreatic αTC1-6 cells, which act as an attractive model system for epigenetic cell reprogramming and diabetes treatment, were transiently transfected with two different transfection methods: the chemical method with polyethyleneimine (PEI) and nucleofection as a physical transfection method. Flow cytometry and fluorescent microscopy examination of GFP expression showed that transfection efficiency was affected by the size of plasmids using both transfection methods. Subsequently, the Cas9 mRNA expression confirmed successful transfection with EpiCRISPR plasmid, whereas the cell physiology remained unchanged. The adjusted nucleofection protocol for αTC1-6 cells transfected with an EpiCRISPR mix of plasmids reached 71.1% of GFP-positive transfected cells on the fifth post-transfection day and proved to be much more efficient than the 3.8% GFP-positive PEI transfected cells. Modifying the protocol, we finally specify CM-156 program and SF 4D-Nucleofector X Solutions for Amaxa™ nucleofection as a method of choice for alpha TC1-6 cell line transfection.
PB  - Basel: MDPI
T2  - Applied Sciences
T1  - Nucleofection as an Efficient Method for Alpha TC1-6 Cell Line Transfection
IS  - 15
VL  - 12
DO  - 10.3390/app12157938
SP  - 7938
ER  - 

@article{
author = "Đorđević, Marija and Paunović, Verica and Jovanović Tucović, Maja and Tolić, Anja and Rajić, Jovana and Dinić, Svetlana and Uskoković, Aleksandra and Grdović, Nevena and Mihailović, Mirjana and Marković, Ivanka and Arambašić Jovanović, Jelena and Vidaković, Melita",
year = "2022",
abstract = "An efficient transfection is a crucial step for the introduction of epigenetic modification in host cells, and there is a need for an optimized transfection process for individual model systems separately. Mouse pancreatic αTC1-6 cells, which act as an attractive model system for epigenetic cell reprogramming and diabetes treatment, were transiently transfected with two different transfection methods: the chemical method with polyethyleneimine (PEI) and nucleofection as a physical transfection method. Flow cytometry and fluorescent microscopy examination of GFP expression showed that transfection efficiency was affected by the size of plasmids using both transfection methods. Subsequently, the Cas9 mRNA expression confirmed successful transfection with EpiCRISPR plasmid, whereas the cell physiology remained unchanged. The adjusted nucleofection protocol for αTC1-6 cells transfected with an EpiCRISPR mix of plasmids reached 71.1% of GFP-positive transfected cells on the fifth post-transfection day and proved to be much more efficient than the 3.8% GFP-positive PEI transfected cells. Modifying the protocol, we finally specify CM-156 program and SF 4D-Nucleofector X Solutions for Amaxa™ nucleofection as a method of choice for alpha TC1-6 cell line transfection.",
publisher = "Basel: MDPI",
journal = "Applied Sciences",
title = "Nucleofection as an Efficient Method for Alpha TC1-6 Cell Line Transfection",
number = "15",
volume = "12",
doi = "10.3390/app12157938",
pages = "7938"
}

Đorđević, M., Paunović, V., Jovanović Tucović, M., Tolić, A., Rajić, J., Dinić, S., Uskoković, A., Grdović, N., Mihailović, M., Marković, I., Arambašić Jovanović, J.,& Vidaković, M.. (2022). Nucleofection as an Efficient Method for Alpha TC1-6 Cell Line Transfection. in Applied Sciences
Basel: MDPI., 12(15), 7938.
https://doi.org/10.3390/app12157938

Đorđević M, Paunović V, Jovanović Tucović M, Tolić A, Rajić J, Dinić S, Uskoković A, Grdović N, Mihailović M, Marković I, Arambašić Jovanović J, Vidaković M. Nucleofection as an Efficient Method for Alpha TC1-6 Cell Line Transfection. in Applied Sciences. 2022;12(15):7938.
doi:10.3390/app12157938 .

Đorđević, Marija, Paunović, Verica, Jovanović Tucović, Maja, Tolić, Anja, Rajić, Jovana, Dinić, Svetlana, Uskoković, Aleksandra, Grdović, Nevena, Mihailović, Mirjana, Marković, Ivanka, Arambašić Jovanović, Jelena, Vidaković, Melita, "Nucleofection as an Efficient Method for Alpha TC1-6 Cell Line Transfection" in Applied Sciences, 12, no. 15 (2022):7938,
https://doi.org/10.3390/app12157938 . .

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  - Harhaji-Trajković, Ljubica
AU  - Kosić, Milica
AU  - Paunović, Verica
AU  - Ristić, Biljana
AU  - Bošnjak, Mihajlo
AU  - Zogović, Nevena
AU  - Mandić, Miloš
AU  - Tovilović-Kovačević, Gordana
AU  - Janjetović, Kristina
AU  - Trajković, Vladimir
PY  - 2021
UR  - https://www.sdir.ac.rs/apstrakti-SDIR-5/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4709
AB  - Background: Intensive proliferation of tumor cells consumes a lot of energy. In nutrient deficiency 
substrates for energy metabolism are obtained by lysosomal degradation of unnecessary/dysfunctional 
intracellular organelles/molecules in the process of autophagy. Leakage of enlarged unstable lysosomes, 
which characterize tumor cells, causes cell death. We investigated antitumor effect of combined targeting 
of lysosomes/autophagy and energy metabolism. Material and Methods: Toxicity against U251 human 
glioma and B16 mouse melanoma cells was measured by viability tests. Type/mechanisms of cell death 
were determined by flow cytometry, immunoblot, fluorescent/electron microscopy and confirmed by 
appropriate genetic/pharmacological inhibitors. Therapeutic potential was estimated in B16 melanoma bearing C57Bl/6 mice. Results: In the first study, lysosomotropic autophagy inhibitor chloroquine (CQ) 
rapidly killed tumor cells incubated in the absence of serum. CQ-induced lysosomal destabilization 
triggered: oxidative stress, mitochondrial depolarization, and mixed apoptosis/necrosis of serum-deprived 
cells. In the second study, lysosomal detergent N-dodecylimidazole (NDI) synergized in antitumor activity 
with the glycolytic inhibitor 2-deoxy-D-glucose (2DG). NDI-triggered release of lysosomal enzymes into the 
cytoplasm caused mitochondrial damage and blocked oxidative phosphorylation, which synergized with 
2DG-mediated glycolysis block in ATP reduction, oxidative stress, and necrosis. Interestingly, although both 
serum deprivation and 2DG stimulated autophagy, CQ- and NDI-induced autophagy suppression was 
irrelevant for their cytotoxicity. Importantly, CQ+food restriction and 2DG+NDI reduced melanoma growth 
in vivo. Conclusion: Autophagy independent antitumor effects of combined energy metabolism suppression 
and lysosomal destabilization might be exploited in cancer therapy.
PB  - Beograd : Srpsko društvo istraživača raka
C3  - 5th Congress of the Serbian Association for Cancer Research – SDIR-5 with international participation „Translational potential of cancer research in Serbia“
T1  - Dual targeting of energy metabolism and lysosomes as an anticancer strategy; It is not all about autophagy
SP  - 8
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4709
ER  - 

@conference{
author = "Harhaji-Trajković, Ljubica and Kosić, Milica and Paunović, Verica and Ristić, Biljana and Bošnjak, Mihajlo and Zogović, Nevena and Mandić, Miloš and Tovilović-Kovačević, Gordana and Janjetović, Kristina and Trajković, Vladimir",
year = "2021",
abstract = "Background: Intensive proliferation of tumor cells consumes a lot of energy. In nutrient deficiency 
substrates for energy metabolism are obtained by lysosomal degradation of unnecessary/dysfunctional 
intracellular organelles/molecules in the process of autophagy. Leakage of enlarged unstable lysosomes, 
which characterize tumor cells, causes cell death. We investigated antitumor effect of combined targeting 
of lysosomes/autophagy and energy metabolism. Material and Methods: Toxicity against U251 human 
glioma and B16 mouse melanoma cells was measured by viability tests. Type/mechanisms of cell death 
were determined by flow cytometry, immunoblot, fluorescent/electron microscopy and confirmed by 
appropriate genetic/pharmacological inhibitors. Therapeutic potential was estimated in B16 melanoma bearing C57Bl/6 mice. Results: In the first study, lysosomotropic autophagy inhibitor chloroquine (CQ) 
rapidly killed tumor cells incubated in the absence of serum. CQ-induced lysosomal destabilization 
triggered: oxidative stress, mitochondrial depolarization, and mixed apoptosis/necrosis of serum-deprived 
cells. In the second study, lysosomal detergent N-dodecylimidazole (NDI) synergized in antitumor activity 
with the glycolytic inhibitor 2-deoxy-D-glucose (2DG). NDI-triggered release of lysosomal enzymes into the 
cytoplasm caused mitochondrial damage and blocked oxidative phosphorylation, which synergized with 
2DG-mediated glycolysis block in ATP reduction, oxidative stress, and necrosis. Interestingly, although both 
serum deprivation and 2DG stimulated autophagy, CQ- and NDI-induced autophagy suppression was 
irrelevant for their cytotoxicity. Importantly, CQ+food restriction and 2DG+NDI reduced melanoma growth 
in vivo. Conclusion: Autophagy independent antitumor effects of combined energy metabolism suppression 
and lysosomal destabilization might be exploited in cancer therapy.",
publisher = "Beograd : Srpsko društvo istraživača raka",
journal = "5th Congress of the Serbian Association for Cancer Research – SDIR-5 with international participation „Translational potential of cancer research in Serbia“",
title = "Dual targeting of energy metabolism and lysosomes as an anticancer strategy; It is not all about autophagy",
pages = "8",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4709"
}

Harhaji-Trajković, L., Kosić, M., Paunović, V., Ristić, B., Bošnjak, M., Zogović, N., Mandić, M., Tovilović-Kovačević, G., Janjetović, K.,& Trajković, V.. (2021). Dual targeting of energy metabolism and lysosomes as an anticancer strategy; It is not all about autophagy. in 5th Congress of the Serbian Association for Cancer Research – SDIR-5 with international participation „Translational potential of cancer research in Serbia“
Beograd : Srpsko društvo istraživača raka., 8.
https://hdl.handle.net/21.15107/rcub_ibiss_4709

Harhaji-Trajković L, Kosić M, Paunović V, Ristić B, Bošnjak M, Zogović N, Mandić M, Tovilović-Kovačević G, Janjetović K, Trajković V. Dual targeting of energy metabolism and lysosomes as an anticancer strategy; It is not all about autophagy. in 5th Congress of the Serbian Association for Cancer Research – SDIR-5 with international participation „Translational potential of cancer research in Serbia“. 2021;:8.
https://hdl.handle.net/21.15107/rcub_ibiss_4709 .

Harhaji-Trajković, Ljubica, Kosić, Milica, Paunović, Verica, Ristić, Biljana, Bošnjak, Mihajlo, Zogović, Nevena, Mandić, Miloš, Tovilović-Kovačević, Gordana, Janjetović, Kristina, Trajković, Vladimir, "Dual targeting of energy metabolism and lysosomes as an anticancer strategy; It is not all about autophagy" in 5th Congress of the Serbian Association for Cancer Research – SDIR-5 with international participation „Translational potential of cancer research in Serbia“ (2021):8,
https://hdl.handle.net/21.15107/rcub_ibiss_4709 .

TY  - CONF
AU  - Paunović, Verica
AU  - Kosić, Milica
AU  - Ristić, Biljana
AU  - Bošnjak, Mihajlo
AU  - Stevanović, Danijela
AU  - Misirkić Marjanović, Maja
AU  - Mandić, Miloš
AU  - Mirčić, Aleksandar
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
PY  - 2021
UR  - https://www.sfrre2021belgrade.rs/
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4728
AB  - We investigated the effect of 3-methyladenine (3MA), a class III phosphatidylinositol 3-
kinase (PI3K)-blocking autophagy inhibitor, on the melanoma cell death induced by simultaneous
inhibition of glycolysis by 2-deoxyglucose (2DG) and mitochondrial respiration by rotenone. We
have elsewhere shown that 2DG/rotenone caused oxidative stress, ATP depletion, swelling
of mitochondria, ultimately leading to necrosis. Energy stress is known to induce autophagy,
a tightly regulated self-degradation process, which by recycling damaged organelles and
macromolecules provides building blocks and energy. However, 2DG/rotenone did not induce
proautophagic beclin-1 expression and autophagic flux in melanoma cells despite activation
of AMP-activated protein kinase (AMPK) and subsequent inhibition of mammalian target of
rapamycin complex 1 (mTORC1). 3MA, but not autophagy inhibition with other PI3K and lysosomal
inhibitors, attenuated 2DG/rotenone-induced mitochondrial damage, oxidative stress, ATP
depletion, and cell death. 3MA increased both AMPK and mTORC1 activation in energy stressed
cells, but neither AMPK nor mTORC1 inhibition reduced its cytoprotective effect. 3MA reduced
superoxide generation and c-Jun N-terminal kinase (JNK) activation, and both antioxidant and
JNK blockade mimicked its protective activity. Therefore, 3MA prevents energy stress-triggered
melanoma cell death through autophagy-independent decrease of oxidative stress and JNK
activation. Our results warrant caution in use of 3MA as an autophagy inhibitor.
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  - 3-methyladenine protects melanoma cells against energy stress-induced necrosis by autophagy-independent decrease in oxidative stress and partial involvement of JNK
DO  - 10.1016/j.freeradbiomed.2021.08.223
SP  - 221
ER  - 

@conference{
author = "Paunović, Verica and Kosić, Milica and Ristić, Biljana and Bošnjak, Mihajlo and Stevanović, Danijela and Misirkić Marjanović, Maja and Mandić, Miloš and Mirčić, Aleksandar and Trajković, Vladimir and Harhaji-Trajković, Ljubica",
year = "2021",
abstract = "We investigated the effect of 3-methyladenine (3MA), a class III phosphatidylinositol 3-
kinase (PI3K)-blocking autophagy inhibitor, on the melanoma cell death induced by simultaneous
inhibition of glycolysis by 2-deoxyglucose (2DG) and mitochondrial respiration by rotenone. We
have elsewhere shown that 2DG/rotenone caused oxidative stress, ATP depletion, swelling
of mitochondria, ultimately leading to necrosis. Energy stress is known to induce autophagy,
a tightly regulated self-degradation process, which by recycling damaged organelles and
macromolecules provides building blocks and energy. However, 2DG/rotenone did not induce
proautophagic beclin-1 expression and autophagic flux in melanoma cells despite activation
of AMP-activated protein kinase (AMPK) and subsequent inhibition of mammalian target of
rapamycin complex 1 (mTORC1). 3MA, but not autophagy inhibition with other PI3K and lysosomal
inhibitors, attenuated 2DG/rotenone-induced mitochondrial damage, oxidative stress, ATP
depletion, and cell death. 3MA increased both AMPK and mTORC1 activation in energy stressed
cells, but neither AMPK nor mTORC1 inhibition reduced its cytoprotective effect. 3MA reduced
superoxide generation and c-Jun N-terminal kinase (JNK) activation, and both antioxidant and
JNK blockade mimicked its protective activity. Therefore, 3MA prevents energy stress-triggered
melanoma cell death through autophagy-independent decrease of oxidative stress and JNK
activation. Our results warrant caution in use of 3MA as an autophagy inhibitor.",
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 = "3-methyladenine protects melanoma cells against energy stress-induced necrosis by autophagy-independent decrease in oxidative stress and partial involvement of JNK",
doi = "10.1016/j.freeradbiomed.2021.08.223",
pages = "221"
}

Paunović, V., Kosić, M., Ristić, B., Bošnjak, M., Stevanović, D., Misirkić Marjanović, M., Mandić, M., Mirčić, A., Trajković, V.,& Harhaji-Trajković, L.. (2021). 3-methyladenine protects melanoma cells against energy stress-induced necrosis by autophagy-independent decrease in oxidative stress and partial involvement of JNK. 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.., 221.
https://doi.org/10.1016/j.freeradbiomed.2021.08.223

Paunović V, Kosić M, Ristić B, Bošnjak M, Stevanović D, Misirkić Marjanović M, Mandić M, Mirčić A, Trajković V, Harhaji-Trajković L. 3-methyladenine protects melanoma cells against energy stress-induced necrosis by autophagy-independent decrease in oxidative stress and partial involvement of JNK. 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;:221.
doi:10.1016/j.freeradbiomed.2021.08.223 .

Paunović, Verica, Kosić, Milica, Ristić, Biljana, Bošnjak, Mihajlo, Stevanović, Danijela, Misirkić Marjanović, Maja, Mandić, Miloš, Mirčić, Aleksandar, Trajković, Vladimir, Harhaji-Trajković, Ljubica, "3-methyladenine protects melanoma cells against energy stress-induced necrosis by autophagy-independent decrease in oxidative stress and partial involvement of JNK" 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):221,
https://doi.org/10.1016/j.freeradbiomed.2021.08.223 . .

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  - Krunić, Matija
AU  - Ristić, Biljana
AU  - Bošnjak, Mihajlo
AU  - Paunović, Verica
AU  - Tovilović-Kovačević, Gordana
AU  - Zogović, Nevena
AU  - Mirčić, Aleksandar
AU  - Marković, Zoran
AU  - Todorović-Marković, Biljana
AU  - Jovanović, Svetlana
AU  - Kleut, Duška
AU  - Mojović, Miloš
AU  - Nakarada, Đura
AU  - Marković, Olivera
AU  - Vuković, Irena
AU  - Harhaji-Trajković, Ljubica
AU  - Trajković, Vladimir
PY  - 2021
UR  - https://linkinghub.elsevier.com/retrieve/pii/S0891584921007760
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4655
AB  - We investigated the ability of graphene quantum dot (GQD) nanoparticles to protect SH-SY5Y human neuroblastoma cells from oxidative/nitrosative stress induced by iron-nitrosyl complex sodium nitroprusside (SNP). GQD reduced SNP cytotoxicity by preventing mitochondrial depolarization, caspase-2 activation, and subsequent apoptotic death. Although GQD diminished the levels of nitric oxide (NO) in SNP-exposed cells, NO scavengers displayed only a slight protective effect, suggesting that NO quenching was not the main protective mechanism of GQD. GQD also reduced SNP-triggered increase in the intracellular levels of hydroxyl radical (•OH), superoxide anion (O2•-), and lipid peroxidation. Nonselective antioxidants, •OH scavenging, and iron chelators, but not superoxide dismutase, mimicked GQD cytoprotective activity, indicating that GQD protect cells by neutralizing •OH generated in the presence of SNP-released iron. Cellular internalization of GQD was required for optimal protection, since a removal of extracellular GQD by extensive washing only partly diminished their protective effect. Moreover, GQD cooperated with SNP to induce autophagy, as confirmed by the inhibition of autophagy-limiting Akt/PRAS40/mTOR signaling and increase in autophagy gene transcription, protein levels of proautophagic beclin-1 and LC3-II, formation of autophagic vesicles, and degradation of autophagic target p62. The antioxidant activity of GQD was not involved in autophagy induction, as antioxidants N-acetylcysteine and dimethyl sulfoxide failed to stimulate autophagy in SNP-exposed cells. Pharmacological inhibitors of early (wortmannin, 3-methyladenine) or late stages of autophagy (NH4Cl) efficiently reduced the protective effect of GQD. Therefore, the ability of GQD to prevent the in vitro neurotoxicity of SNP depends on both •OH/NO scavenging and induction of cytoprotective autophagy.
PB  - Elsevier Inc.
T2  - Free Radical Biology and Medicine
T1  - Graphene quantum dot antioxidant and proautophagic actions protect SH-SY5Y neuroblastoma cells from oxidative stress-mediated apoptotic death.
VL  - 177
DO  - 10.1016/j.freeradbiomed.2021.10.025
SP  - 167
EP  - 180
ER  - 

@article{
author = "Krunić, Matija and Ristić, Biljana and Bošnjak, Mihajlo and Paunović, Verica and Tovilović-Kovačević, Gordana and Zogović, Nevena and Mirčić, Aleksandar and Marković, Zoran and Todorović-Marković, Biljana and Jovanović, Svetlana and Kleut, Duška and Mojović, Miloš and Nakarada, Đura and Marković, Olivera and Vuković, Irena and Harhaji-Trajković, Ljubica and Trajković, Vladimir",
year = "2021",
abstract = "We investigated the ability of graphene quantum dot (GQD) nanoparticles to protect SH-SY5Y human neuroblastoma cells from oxidative/nitrosative stress induced by iron-nitrosyl complex sodium nitroprusside (SNP). GQD reduced SNP cytotoxicity by preventing mitochondrial depolarization, caspase-2 activation, and subsequent apoptotic death. Although GQD diminished the levels of nitric oxide (NO) in SNP-exposed cells, NO scavengers displayed only a slight protective effect, suggesting that NO quenching was not the main protective mechanism of GQD. GQD also reduced SNP-triggered increase in the intracellular levels of hydroxyl radical (•OH), superoxide anion (O2•-), and lipid peroxidation. Nonselective antioxidants, •OH scavenging, and iron chelators, but not superoxide dismutase, mimicked GQD cytoprotective activity, indicating that GQD protect cells by neutralizing •OH generated in the presence of SNP-released iron. Cellular internalization of GQD was required for optimal protection, since a removal of extracellular GQD by extensive washing only partly diminished their protective effect. Moreover, GQD cooperated with SNP to induce autophagy, as confirmed by the inhibition of autophagy-limiting Akt/PRAS40/mTOR signaling and increase in autophagy gene transcription, protein levels of proautophagic beclin-1 and LC3-II, formation of autophagic vesicles, and degradation of autophagic target p62. The antioxidant activity of GQD was not involved in autophagy induction, as antioxidants N-acetylcysteine and dimethyl sulfoxide failed to stimulate autophagy in SNP-exposed cells. Pharmacological inhibitors of early (wortmannin, 3-methyladenine) or late stages of autophagy (NH4Cl) efficiently reduced the protective effect of GQD. Therefore, the ability of GQD to prevent the in vitro neurotoxicity of SNP depends on both •OH/NO scavenging and induction of cytoprotective autophagy.",
publisher = "Elsevier Inc.",
journal = "Free Radical Biology and Medicine",
title = "Graphene quantum dot antioxidant and proautophagic actions protect SH-SY5Y neuroblastoma cells from oxidative stress-mediated apoptotic death.",
volume = "177",
doi = "10.1016/j.freeradbiomed.2021.10.025",
pages = "167-180"
}

Krunić, M., Ristić, B., Bošnjak, M., Paunović, V., Tovilović-Kovačević, G., Zogović, N., Mirčić, A., Marković, Z., Todorović-Marković, B., Jovanović, S., Kleut, D., Mojović, M., Nakarada, Đ., Marković, O., Vuković, I., Harhaji-Trajković, L.,& Trajković, V.. (2021). Graphene quantum dot antioxidant and proautophagic actions protect SH-SY5Y neuroblastoma cells from oxidative stress-mediated apoptotic death.. in Free Radical Biology and Medicine
Elsevier Inc.., 177, 167-180.
https://doi.org/10.1016/j.freeradbiomed.2021.10.025

Krunić M, Ristić B, Bošnjak M, Paunović V, Tovilović-Kovačević G, Zogović N, Mirčić A, Marković Z, Todorović-Marković B, Jovanović S, Kleut D, Mojović M, Nakarada Đ, Marković O, Vuković I, Harhaji-Trajković L, Trajković V. Graphene quantum dot antioxidant and proautophagic actions protect SH-SY5Y neuroblastoma cells from oxidative stress-mediated apoptotic death.. in Free Radical Biology and Medicine. 2021;177:167-180.
doi:10.1016/j.freeradbiomed.2021.10.025 .

Krunić, Matija, Ristić, Biljana, Bošnjak, Mihajlo, Paunović, Verica, Tovilović-Kovačević, Gordana, Zogović, Nevena, Mirčić, Aleksandar, Marković, Zoran, Todorović-Marković, Biljana, Jovanović, Svetlana, Kleut, Duška, Mojović, Miloš, Nakarada, Đura, Marković, Olivera, Vuković, Irena, Harhaji-Trajković, Ljubica, Trajković, Vladimir, "Graphene quantum dot antioxidant and proautophagic actions protect SH-SY5Y neuroblastoma cells from oxidative stress-mediated apoptotic death." in Free Radical Biology and Medicine, 177 (2021):167-180,
https://doi.org/10.1016/j.freeradbiomed.2021.10.025 . .

TY  - JOUR
AU  - Kosić, Milica
AU  - Paunović, Verica
AU  - Ristić, Biljana
AU  - Mirčić, Aleksandar
AU  - Bošnjak, Mihajlo
AU  - Stevanović, Danijela
AU  - Kravić-Stevović, Tamara K
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S1347861321000591
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4427
AB  - We investigated the effect of 3-methyladenine (3MA), a class III phosphatidylinositol 3-kinase (PI3K)-blocking autophagy inhibitor, on cancer cell death induced by simultaneous inhibition of glycolysis by 2-deoxyglucose (2DG) and mitochondrial respiration by rotenone. 2DG/rotenone reduced ATP levels and increased mitochondrial superoxide production, causing mitochondrial swelling and necrotic death in various cancer cell lines. 2DG/rotenone failed to increase proautophagic beclin-1 and autophagic flux in melanoma cells despite the activation of AMP-activated protein kinase (AMPK) and inhibition of mechanistic target of rapamycin complex 1 (mTORC1). 3MA, but not autophagy inhibition with other PI3K and lysosomal inhibitors, attenuated 2DG/rotenone-induced mitochondrial damage, oxidative stress, ATP depletion, and cell death, while antioxidant treatment mimicked its protective action. The protection was not mediated by autophagy upregulation via class I PI3K/Akt inhibition, as it was preserved in cells with genetically inhibited autophagy. 3MA increased AMPK and mTORC1 activation in energy-stressed cells, but neither AMPK nor mTORC1 inhibition reduced its cytoprotective effect. 3MA reduced JNK activation, and JNK pharmacological/genetic suppression mimicked its mitochondria-preserving and cytoprotective activity. Therefore, 3MA prevents energy stress-triggered cancer cell death through autophagy-independent mechanisms possibly involving JNK suppression and decrease of oxidative stress. Our results warrant caution when using 3MA as an autophagy inhibitor.
PB  - Kyoto : Japanese Pharmacological Society
T2  - Journal of Pharmacological Sciences
T1  - 3-Methyladenine prevents energy stress-induced necrotic death of melanoma cells through autophagy-independent mechanisms
IS  - 1
VL  - 147
DO  - 10.1016/j.jphs.2021.06.003
SP  - 156
EP  - 167
ER  - 

@article{
author = "Kosić, Milica and Paunović, Verica and Ristić, Biljana and Mirčić, Aleksandar and Bošnjak, Mihajlo and Stevanović, Danijela and Kravić-Stevović, Tamara K and Trajković, Vladimir and Harhaji-Trajković, Ljubica",
year = "2021",
abstract = "We investigated the effect of 3-methyladenine (3MA), a class III phosphatidylinositol 3-kinase (PI3K)-blocking autophagy inhibitor, on cancer cell death induced by simultaneous inhibition of glycolysis by 2-deoxyglucose (2DG) and mitochondrial respiration by rotenone. 2DG/rotenone reduced ATP levels and increased mitochondrial superoxide production, causing mitochondrial swelling and necrotic death in various cancer cell lines. 2DG/rotenone failed to increase proautophagic beclin-1 and autophagic flux in melanoma cells despite the activation of AMP-activated protein kinase (AMPK) and inhibition of mechanistic target of rapamycin complex 1 (mTORC1). 3MA, but not autophagy inhibition with other PI3K and lysosomal inhibitors, attenuated 2DG/rotenone-induced mitochondrial damage, oxidative stress, ATP depletion, and cell death, while antioxidant treatment mimicked its protective action. The protection was not mediated by autophagy upregulation via class I PI3K/Akt inhibition, as it was preserved in cells with genetically inhibited autophagy. 3MA increased AMPK and mTORC1 activation in energy-stressed cells, but neither AMPK nor mTORC1 inhibition reduced its cytoprotective effect. 3MA reduced JNK activation, and JNK pharmacological/genetic suppression mimicked its mitochondria-preserving and cytoprotective activity. Therefore, 3MA prevents energy stress-triggered cancer cell death through autophagy-independent mechanisms possibly involving JNK suppression and decrease of oxidative stress. Our results warrant caution when using 3MA as an autophagy inhibitor.",
publisher = "Kyoto : Japanese Pharmacological Society",
journal = "Journal of Pharmacological Sciences",
title = "3-Methyladenine prevents energy stress-induced necrotic death of melanoma cells through autophagy-independent mechanisms",
number = "1",
volume = "147",
doi = "10.1016/j.jphs.2021.06.003",
pages = "156-167"
}

Kosić, M., Paunović, V., Ristić, B., Mirčić, A., Bošnjak, M., Stevanović, D., Kravić-Stevović, T. K., Trajković, V.,& Harhaji-Trajković, L.. (2021). 3-Methyladenine prevents energy stress-induced necrotic death of melanoma cells through autophagy-independent mechanisms. in Journal of Pharmacological Sciences
Kyoto : Japanese Pharmacological Society., 147(1), 156-167.
https://doi.org/10.1016/j.jphs.2021.06.003

Kosić M, Paunović V, Ristić B, Mirčić A, Bošnjak M, Stevanović D, Kravić-Stevović TK, Trajković V, Harhaji-Trajković L. 3-Methyladenine prevents energy stress-induced necrotic death of melanoma cells through autophagy-independent mechanisms. in Journal of Pharmacological Sciences. 2021;147(1):156-167.
doi:10.1016/j.jphs.2021.06.003 .

Kosić, Milica, Paunović, Verica, Ristić, Biljana, Mirčić, Aleksandar, Bošnjak, Mihajlo, Stevanović, Danijela, Kravić-Stevović, Tamara K, Trajković, Vladimir, Harhaji-Trajković, Ljubica, "3-Methyladenine prevents energy stress-induced necrotic death of melanoma cells through autophagy-independent mechanisms" in Journal of Pharmacological Sciences, 147, no. 1 (2021):156-167,
https://doi.org/10.1016/j.jphs.2021.06.003 . .

TY  - JOUR
AU  - Paunović, Verica
AU  - Kosić, Milica
AU  - Misirkić Marjanović, Maja
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
PY  - 2020
UR  - http://www.ncbi.nlm.nih.gov/pubmed/33383091
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4096
AB  - To sustain their proliferative and metastatic capacity, tumor cells increase the activity of energy-producing pathways and lysosomal compartment, resorting to autophagolysosomal degradation when nutrients are scarce. Consequently, large fragile lysosomes and enhanced energy metabolism may serve as targets for anticancer therapy. A simultaneous induction of energy stress (by caloric restriction and inhibition of glycolysis, oxidative phosphorylation, Krebs cycle, or amino acid/fatty acid metabolism) and lysosomal stress (by lysosomotropic detergents, vacuolar ATPase inhibitors, or cationic amphiphilic drugs) is an efficient anti-cancer strategy demonstrated in a number of studies. However, the mechanisms of lysosomal/energy stress co-amplification, apart from the protective autophagy inhibition, are poorly understood. We here summarize the established and suggest potential mechanisms and candidates for anticancer therapy based on the dual targeting of lysosomes and energy metabolism.
PB  - Elsevier BV
T2  - Biochimica et Biophysica Acta (BBA) - Molecular Cell Research
T1  - Dual targeting of tumor cell energy metabolism and lysosomes as an anticancer strategy.
IS  - 4
VL  - 1868
DO  - 10.1016/j.bbamcr.2020.118944
SP  - 118944
ER  - 

@article{
author = "Paunović, Verica and Kosić, Milica and Misirkić Marjanović, Maja and Trajković, Vladimir and Harhaji-Trajković, Ljubica",
year = "2020",
abstract = "To sustain their proliferative and metastatic capacity, tumor cells increase the activity of energy-producing pathways and lysosomal compartment, resorting to autophagolysosomal degradation when nutrients are scarce. Consequently, large fragile lysosomes and enhanced energy metabolism may serve as targets for anticancer therapy. A simultaneous induction of energy stress (by caloric restriction and inhibition of glycolysis, oxidative phosphorylation, Krebs cycle, or amino acid/fatty acid metabolism) and lysosomal stress (by lysosomotropic detergents, vacuolar ATPase inhibitors, or cationic amphiphilic drugs) is an efficient anti-cancer strategy demonstrated in a number of studies. However, the mechanisms of lysosomal/energy stress co-amplification, apart from the protective autophagy inhibition, are poorly understood. We here summarize the established and suggest potential mechanisms and candidates for anticancer therapy based on the dual targeting of lysosomes and energy metabolism.",
publisher = "Elsevier BV",
journal = "Biochimica et Biophysica Acta (BBA) - Molecular Cell Research",
title = "Dual targeting of tumor cell energy metabolism and lysosomes as an anticancer strategy.",
number = "4",
volume = "1868",
doi = "10.1016/j.bbamcr.2020.118944",
pages = "118944"
}

Paunović, V., Kosić, M., Misirkić Marjanović, M., Trajković, V.,& Harhaji-Trajković, L.. (2020). Dual targeting of tumor cell energy metabolism and lysosomes as an anticancer strategy.. in Biochimica et Biophysica Acta (BBA) - Molecular Cell Research
Elsevier BV., 1868(4), 118944.
https://doi.org/10.1016/j.bbamcr.2020.118944

Paunović V, Kosić M, Misirkić Marjanović M, Trajković V, Harhaji-Trajković L. Dual targeting of tumor cell energy metabolism and lysosomes as an anticancer strategy.. in Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 2020;1868(4):118944.
doi:10.1016/j.bbamcr.2020.118944 .

Paunović, Verica, Kosić, Milica, Misirkić Marjanović, Maja, Trajković, Vladimir, Harhaji-Trajković, Ljubica, "Dual targeting of tumor cell energy metabolism and lysosomes as an anticancer strategy." in Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1868, no. 4 (2020):118944,
https://doi.org/10.1016/j.bbamcr.2020.118944 . .

TY  - JOUR
AU  - Koprivica, Ivan
AU  - Mićanović, Dragica
AU  - Pejnović, Nada
AU  - Paunović, Verica
AU  - Saksida, Tamara
AU  - Stojanović, Ivana D.
PY  - 2020
UR  - https://www.mdpi.com/1420-3049/25/18/4112
UR  - http://www.ncbi.nlm.nih.gov/pubmed/32916780
UR  - https://radar.ibiss.bg.ac.rs/123456789/3892
AB  - Ethyl pyruvate (EP), a stable form of pyruvate, has shown beneficial effects in animal models of shock, ischemia/reperfusion injury, and sepsis due to its potent anti-oxidant and anti-inflammatory properties. Our recent study demonstrated that EP application prevented the clinical manifestation of type 1 diabetes in mice by augmenting regulatory T cell (Treg) number and function. Our present study shows that EP increases Treg proliferation and suppressive function (perforin and IL-10 expression) during in vitro differentiation from conventional CD4+CD25- T cells. Enhanced expansion of Treg after EP treatment correlated with increased ATP levels and relied on increased glycolysis. Inhibition of oxidative phosphorylation did not attenuate EP stimulatory effects, suggesting that this metabolic pathway was not mandatory for EP-driven Treg proliferation. Moreover, EP lowered the expression of carnitine palmitoyltransferase I, an enzyme involved in fatty acid oxidation. Further, the stimulatory effect of EP on Treg proliferation was not mediated through inhibition of the mTOR signaling pathway. When given in vivo either intraperitoneally or orally to healthy C57BL/6 mice, EP increased the number of Treg within the peritoneal cavity or gut-associated lymphoid tissue, respectively. In conclusion, EP promotes in vitro Treg proliferation through increased glycolysis and enhances Treg proliferation when administered in vivo.
PB  - MDPI AG
T2  - Molecules (Basel, Switzerland)
T1  - Ethyl Pyruvate Promotes Proliferation of Regulatory T Cells by Increasing Glycolysis.
IS  - 18
VL  - 25
DO  - 10.3390/molecules25184112
SP  - 4112
ER  - 

@article{
author = "Koprivica, Ivan and Mićanović, Dragica and Pejnović, Nada and Paunović, Verica and Saksida, Tamara and Stojanović, Ivana D.",
year = "2020",
abstract = "Ethyl pyruvate (EP), a stable form of pyruvate, has shown beneficial effects in animal models of shock, ischemia/reperfusion injury, and sepsis due to its potent anti-oxidant and anti-inflammatory properties. Our recent study demonstrated that EP application prevented the clinical manifestation of type 1 diabetes in mice by augmenting regulatory T cell (Treg) number and function. Our present study shows that EP increases Treg proliferation and suppressive function (perforin and IL-10 expression) during in vitro differentiation from conventional CD4+CD25- T cells. Enhanced expansion of Treg after EP treatment correlated with increased ATP levels and relied on increased glycolysis. Inhibition of oxidative phosphorylation did not attenuate EP stimulatory effects, suggesting that this metabolic pathway was not mandatory for EP-driven Treg proliferation. Moreover, EP lowered the expression of carnitine palmitoyltransferase I, an enzyme involved in fatty acid oxidation. Further, the stimulatory effect of EP on Treg proliferation was not mediated through inhibition of the mTOR signaling pathway. When given in vivo either intraperitoneally or orally to healthy C57BL/6 mice, EP increased the number of Treg within the peritoneal cavity or gut-associated lymphoid tissue, respectively. In conclusion, EP promotes in vitro Treg proliferation through increased glycolysis and enhances Treg proliferation when administered in vivo.",
publisher = "MDPI AG",
journal = "Molecules (Basel, Switzerland)",
title = "Ethyl Pyruvate Promotes Proliferation of Regulatory T Cells by Increasing Glycolysis.",
number = "18",
volume = "25",
doi = "10.3390/molecules25184112",
pages = "4112"
}

Koprivica, I., Mićanović, D., Pejnović, N., Paunović, V., Saksida, T.,& Stojanović, I. D.. (2020). Ethyl Pyruvate Promotes Proliferation of Regulatory T Cells by Increasing Glycolysis.. in Molecules (Basel, Switzerland)
MDPI AG., 25(18), 4112.
https://doi.org/10.3390/molecules25184112

Koprivica I, Mićanović D, Pejnović N, Paunović V, Saksida T, Stojanović ID. Ethyl Pyruvate Promotes Proliferation of Regulatory T Cells by Increasing Glycolysis.. in Molecules (Basel, Switzerland). 2020;25(18):4112.
doi:10.3390/molecules25184112 .

Koprivica, Ivan, Mićanović, Dragica, Pejnović, Nada, Paunović, Verica, Saksida, Tamara, Stojanović, Ivana D., "Ethyl Pyruvate Promotes Proliferation of Regulatory T Cells by Increasing Glycolysis." in Molecules (Basel, Switzerland), 25, no. 18 (2020):4112,
https://doi.org/10.3390/molecules25184112 . .

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  - Paunović, Verica
AU  - Kosić, Milica
AU  - Arsikin-Csordas, Katarina
AU  - Firestone, Raymond A
AU  - Ristić, Biljana
AU  - Mirčić, Aleksandar
AU  - Petričević, Saša
AU  - Bošnjak, Mihajlo
AU  - Zogović, Nevena
AU  - Mandić, Miloš
AU  - Bumbaširević, Vladimir
AU  - Trajković, Vladimir
AU  - Harhaji-Trajković, Ljubica
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/123456789/6355
AB  - During malignant transformation cells acquire changes in metabolism, signaling pathways as well as organelle content. The preferential use of aerobic glycolysis (Warburg effect), along with the increased number and volume of lysosomes can be viewed as glioma cells’ Achilles heels. In the present study, we aimed to examine the in vitro antiglioma effects of combining lysosomal membrane permeabilization (LMP)-inducing agent N-dodecylimidazole (NDI) with glycolytic inhibitor 2-deoxy-D-glucose (2DG).
NDI-triggered LMP and 2DG-mediated glycolysis block synergistically induced rapid ATP depletion, mitochondrial damage, and reactive oxygen species (ROS) production causing necrotic cell death of U251 glioma cells, but not primary astrocytes. Lysosomal cathepsin inhibitor E64 and antioxidant α-tocopherol partially prevented NDI/2DG-induced glioma cell death, thus implying the involvement of LMP and oxidative stress in the observed cytotoxicity. Likewise, LMP-inducing agent chloroquine
showed synergistic cytotoxic effect with 2DG. Similarly, glucose deprivation as well as other glycolytic inhibitors, iodoacetate and sodium fluoride, synergistically cooperated with NDI, further corroborating that the observed antiglioma effect of the NDI/2DG combined treatment was indeed based on LMP and glycolysis block. Based on these results, we concluded that NDI-triggered LMP caused initial mitochondrial damage, which was further increased by 2DG causing the lack of glycolytic ATP
required to maintain mitochondrial health. This created a positive feedback loop of mitochondrial dysfunction, ATP loss, and ROS production, culminating in necrosis. Therefore, the combination of glycolysis inhibitors and LMP-inducing agents seems promising antiglioma strategy.
PB  - Belgrade: Serbian Neuroscience Society
C3  - Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia
T1  - Synergistic antiglioma action of lysosomal membrane permeabilization and glycolysis inhibition
SP  - 213
EP  - 213
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6355
ER  - 

@conference{
author = "Paunović, Verica and Kosić, Milica and Arsikin-Csordas, Katarina and Firestone, Raymond A and Ristić, Biljana and Mirčić, Aleksandar and Petričević, Saša and Bošnjak, Mihajlo and Zogović, Nevena and Mandić, Miloš and Bumbaširević, Vladimir and Trajković, Vladimir and Harhaji-Trajković, Ljubica",
year = "2019",
abstract = "During malignant transformation cells acquire changes in metabolism, signaling pathways as well as organelle content. The preferential use of aerobic glycolysis (Warburg effect), along with the increased number and volume of lysosomes can be viewed as glioma cells’ Achilles heels. In the present study, we aimed to examine the in vitro antiglioma effects of combining lysosomal membrane permeabilization (LMP)-inducing agent N-dodecylimidazole (NDI) with glycolytic inhibitor 2-deoxy-D-glucose (2DG).
NDI-triggered LMP and 2DG-mediated glycolysis block synergistically induced rapid ATP depletion, mitochondrial damage, and reactive oxygen species (ROS) production causing necrotic cell death of U251 glioma cells, but not primary astrocytes. Lysosomal cathepsin inhibitor E64 and antioxidant α-tocopherol partially prevented NDI/2DG-induced glioma cell death, thus implying the involvement of LMP and oxidative stress in the observed cytotoxicity. Likewise, LMP-inducing agent chloroquine
showed synergistic cytotoxic effect with 2DG. Similarly, glucose deprivation as well as other glycolytic inhibitors, iodoacetate and sodium fluoride, synergistically cooperated with NDI, further corroborating that the observed antiglioma effect of the NDI/2DG combined treatment was indeed based on LMP and glycolysis block. Based on these results, we concluded that NDI-triggered LMP caused initial mitochondrial damage, which was further increased by 2DG causing the lack of glycolytic ATP
required to maintain mitochondrial health. This created a positive feedback loop of mitochondrial dysfunction, ATP loss, and ROS production, culminating in necrosis. Therefore, the combination of glycolysis inhibitors and LMP-inducing agents seems promising antiglioma strategy.",
publisher = "Belgrade: Serbian Neuroscience Society",
journal = "Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia",
title = "Synergistic antiglioma action of lysosomal membrane permeabilization and glycolysis inhibition",
pages = "213-213",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6355"
}

Paunović, V., Kosić, M., Arsikin-Csordas, K., Firestone, R. A., Ristić, B., Mirčić, A., Petričević, S., Bošnjak, M., Zogović, N., Mandić, M., Bumbaširević, V., Trajković, V.,& Harhaji-Trajković, L.. (2019). Synergistic antiglioma action of lysosomal membrane permeabilization and glycolysis inhibition. in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia
Belgrade: Serbian Neuroscience Society., 213-213.
https://hdl.handle.net/21.15107/rcub_ibiss_6355

Paunović V, Kosić M, Arsikin-Csordas K, Firestone RA, Ristić B, Mirčić A, Petričević S, Bošnjak M, Zogović N, Mandić M, Bumbaširević V, Trajković V, Harhaji-Trajković L. Synergistic antiglioma action of lysosomal membrane permeabilization and glycolysis inhibition. in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia. 2019;:213-213.
https://hdl.handle.net/21.15107/rcub_ibiss_6355 .

Paunović, Verica, Kosić, Milica, Arsikin-Csordas, Katarina, Firestone, Raymond A, Ristić, Biljana, Mirčić, Aleksandar, Petričević, Saša, Bošnjak, Mihajlo, Zogović, Nevena, Mandić, Miloš, Bumbaširević, Vladimir, Trajković, Vladimir, Harhaji-Trajković, Ljubica, "Synergistic antiglioma action of lysosomal membrane permeabilization and glycolysis inhibition" in Book of Abstract: Federation of European Neuroscience Societies (FENS) Regional Meeting; 2019 Jul 10-13; Belgrade, Serbia (2019):213-213,
https://hdl.handle.net/21.15107/rcub_ibiss_6355 .

TY  - CONF
AU  - Jevtić, Bojan
AU  - Saksida, Tamara
AU  - Mićanović, Dragica
AU  - Koprivica, Ivan
AU  - Paunović, Verica
AU  - Stojanović, Ivana D.
AU  - Pejnović, Nada
PY  - 2019
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5782
AB  - Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by the imbalance
between the CD4 or CD8 T effector (Teff) cells and the FoxP3+CD4 T regulatory cells
(Tregs) that leads to pancreatic beta-cells destruction causing insulin deficiency.
Environmental factors, diet and microbiome are associated with the recent rise in T1D
incidence. Intestinal immune cells must maintain a tolerogenic response in the gut that
involves the development of Tregs. Recent data show that IL-2-producing type 3 innate
lymphoid cells ILC3s (IL-2+ILC3) in the small intestine are essential for maintaining
FoxP3+ Tregs and oral tolerance to dietary antigens and reveal the previously unknown
direct communication between ILC3s and Treg cells in the gut. We investigated the
frequencies of small intestine lamina propria IL-2+ILC3s and FoxP3+ Tregs during
transition from prediabetes to diabetes in young and old female NOD mice. 20 weeks
old, diabetic NOD mice had higher frequencies of LinnegCD45+RORγt+CD127+ ILC3s
in small intestine lamina propria compared to 4 weeks of age-young NOD mice.
However, the frequencies of IL-2-producing ILC3s and CD4+CD25hiFoxP3+ Tregs
were significantly lower in diabetic NOD mice compared to young, prediabetic mice.
We next investigated how microbiota change before diabetes induction is reflected on
Treg and ILC3 populations. Male C57BL/6 mice were treated with broad spectrum
antibiotics (ABX) for 14 days and then T1D was induced by multiple low doses of
streptozotocin (STZ). Ex vivo cell analyses was done on day 10 after the first STZ
injection. The significantly higher incidence of T1D observed in ABX-treated mice
correlated with significantly lower frequencies of IL-2-producing ILC3s and
FoxP3+Tregs in small intestine lamina propria compared to mice treated with STZ only.
The obtained findings show that the decrease of tolerogenic ILC3s and FoxP3+Tregs in
small intestine is associated with the progression and higher incidence of T1D.
PB  - Belgrade: Institute for Biological Research "Siniša Stanković"– National Institute of Republic of Serbia, University of Belgrade
C3  - Immunology at the Confluence of Multidisciplinary Approaches: abstract book: 2019 Dec 6-8; Belgrade, Serbia
T1  - The decrease of tolerogenic ILC3 and Treg cells in small intestine corelates with the profression of type 1 diabetes in mice
SP  - 102
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5782
ER  - 

@conference{
author = "Jevtić, Bojan and Saksida, Tamara and Mićanović, Dragica and Koprivica, Ivan and Paunović, Verica and Stojanović, Ivana D. and Pejnović, Nada",
year = "2019",
abstract = "Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by the imbalance
between the CD4 or CD8 T effector (Teff) cells and the FoxP3+CD4 T regulatory cells
(Tregs) that leads to pancreatic beta-cells destruction causing insulin deficiency.
Environmental factors, diet and microbiome are associated with the recent rise in T1D
incidence. Intestinal immune cells must maintain a tolerogenic response in the gut that
involves the development of Tregs. Recent data show that IL-2-producing type 3 innate
lymphoid cells ILC3s (IL-2+ILC3) in the small intestine are essential for maintaining
FoxP3+ Tregs and oral tolerance to dietary antigens and reveal the previously unknown
direct communication between ILC3s and Treg cells in the gut. We investigated the
frequencies of small intestine lamina propria IL-2+ILC3s and FoxP3+ Tregs during
transition from prediabetes to diabetes in young and old female NOD mice. 20 weeks
old, diabetic NOD mice had higher frequencies of LinnegCD45+RORγt+CD127+ ILC3s
in small intestine lamina propria compared to 4 weeks of age-young NOD mice.
However, the frequencies of IL-2-producing ILC3s and CD4+CD25hiFoxP3+ Tregs
were significantly lower in diabetic NOD mice compared to young, prediabetic mice.
We next investigated how microbiota change before diabetes induction is reflected on
Treg and ILC3 populations. Male C57BL/6 mice were treated with broad spectrum
antibiotics (ABX) for 14 days and then T1D was induced by multiple low doses of
streptozotocin (STZ). Ex vivo cell analyses was done on day 10 after the first STZ
injection. The significantly higher incidence of T1D observed in ABX-treated mice
correlated with significantly lower frequencies of IL-2-producing ILC3s and
FoxP3+Tregs in small intestine lamina propria compared to mice treated with STZ only.
The obtained findings show that the decrease of tolerogenic ILC3s and FoxP3+Tregs in
small intestine is associated with the progression and higher incidence of T1D.",
publisher = "Belgrade: Institute for Biological Research "Siniša Stanković"– National Institute of Republic of Serbia, University of Belgrade",
journal = "Immunology at the Confluence of Multidisciplinary Approaches: abstract book: 2019 Dec 6-8; Belgrade, Serbia",
title = "The decrease of tolerogenic ILC3 and Treg cells in small intestine corelates with the profression of type 1 diabetes in mice",
pages = "102",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5782"
}

Jevtić, B., Saksida, T., Mićanović, D., Koprivica, I., Paunović, V., Stojanović, I. D.,& Pejnović, N.. (2019). The decrease of tolerogenic ILC3 and Treg cells in small intestine corelates with the profression of type 1 diabetes in mice. in Immunology at the Confluence of Multidisciplinary Approaches: abstract book: 2019 Dec 6-8; Belgrade, Serbia
Belgrade: Institute for Biological Research "Siniša Stanković"– National Institute of Republic of Serbia, University of Belgrade., 102.
https://hdl.handle.net/21.15107/rcub_ibiss_5782

Jevtić B, Saksida T, Mićanović D, Koprivica I, Paunović V, Stojanović ID, Pejnović N. The decrease of tolerogenic ILC3 and Treg cells in small intestine corelates with the profression of type 1 diabetes in mice. in Immunology at the Confluence of Multidisciplinary Approaches: abstract book: 2019 Dec 6-8; Belgrade, Serbia. 2019;:102.
https://hdl.handle.net/21.15107/rcub_ibiss_5782 .

Jevtić, Bojan, Saksida, Tamara, Mićanović, Dragica, Koprivica, Ivan, Paunović, Verica, Stojanović, Ivana D., Pejnović, Nada, "The decrease of tolerogenic ILC3 and Treg cells in small intestine corelates with the profression of type 1 diabetes in mice" in Immunology at the Confluence of Multidisciplinary Approaches: abstract book: 2019 Dec 6-8; Belgrade, Serbia (2019):102,
https://hdl.handle.net/21.15107/rcub_ibiss_5782 .

TY  - JOUR
AU  - Nikolovski, Neda
AU  - Paunović, Verica
AU  - Stojanović, Ivana D.
PY  - 2019
UR  - https://www.sciencedirect.com/science/article/pii/S002217591930047X?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3347
AB  - Polyclonal T regulatory cells (Treg - CD4+CD25+CD127lowFoxp3+) are used in several protocols for the treatment of type 1 diabetes (T1D), multiple sclerosis and graft-versus host disease in clinical trials. However, general opinion is that autoantigen-specific Treg could be more efficient in autoimmunity suppression due to their direct effect on pathogenic autoantigen-specific effector T cells. This study describes isolation and expansion of insulin-specific Treg in vitro. Insulin-specific Treg are uniformly distributed in lymphoid tissues however their number is extremely low. To enrich the proportion of insulin-specific Treg, pure CD4+ cells were co-cultured with insulin B chain peptide-loaded dendritic cells, isolated from mice that develop T1D spontaneously - NOD mice. Insulin-specific CD4+ cell expansion peaked after 48 h of incubation and was in favour of Treg. These cells were then sorted using insulin peptide-loaded MHC class II tetramers and cultured in vitro for 48 h in the presence of TCR stimulators, TGF-β and IL-2. The proportion of gained insulin-specific cells with T regulatory phenotype (CD4+CD25highCD127lowGITR+FoxP3+) was in average between 93% and 97%. These cells have shown potent in vitro suppressive effect on T effector cells, produced IL-10 and TGF-β and expressed PD-1 and CD39. Further proliferation of these insulin-specific Treg required the presence of dendritic cells, anti-CD3 antibody and IL-2. This study provides new, reproducible experimental design for the enrichment and expansion of insulin-specific Treg that can be used for the cell-based therapy of autoimmunity.
T2  - Journal of Immunological Methods
T1  - Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.
VL  - 470
DO  - 10.1016/j.jim.2019.04.011
SP  - 46
EP  - 54
ER  - 

@article{
author = "Nikolovski, Neda and Paunović, Verica and Stojanović, Ivana D.",
year = "2019",
abstract = "Polyclonal T regulatory cells (Treg - CD4+CD25+CD127lowFoxp3+) are used in several protocols for the treatment of type 1 diabetes (T1D), multiple sclerosis and graft-versus host disease in clinical trials. However, general opinion is that autoantigen-specific Treg could be more efficient in autoimmunity suppression due to their direct effect on pathogenic autoantigen-specific effector T cells. This study describes isolation and expansion of insulin-specific Treg in vitro. Insulin-specific Treg are uniformly distributed in lymphoid tissues however their number is extremely low. To enrich the proportion of insulin-specific Treg, pure CD4+ cells were co-cultured with insulin B chain peptide-loaded dendritic cells, isolated from mice that develop T1D spontaneously - NOD mice. Insulin-specific CD4+ cell expansion peaked after 48 h of incubation and was in favour of Treg. These cells were then sorted using insulin peptide-loaded MHC class II tetramers and cultured in vitro for 48 h in the presence of TCR stimulators, TGF-β and IL-2. The proportion of gained insulin-specific cells with T regulatory phenotype (CD4+CD25highCD127lowGITR+FoxP3+) was in average between 93% and 97%. These cells have shown potent in vitro suppressive effect on T effector cells, produced IL-10 and TGF-β and expressed PD-1 and CD39. Further proliferation of these insulin-specific Treg required the presence of dendritic cells, anti-CD3 antibody and IL-2. This study provides new, reproducible experimental design for the enrichment and expansion of insulin-specific Treg that can be used for the cell-based therapy of autoimmunity.",
journal = "Journal of Immunological Methods",
title = "Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.",
volume = "470",
doi = "10.1016/j.jim.2019.04.011",
pages = "46-54"
}

Nikolovski, N., Paunović, V.,& Stojanović, I. D.. (2019). Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.. in Journal of Immunological Methods, 470, 46-54.
https://doi.org/10.1016/j.jim.2019.04.011

Nikolovski N, Paunović V, Stojanović ID. Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.. in Journal of Immunological Methods. 2019;470:46-54.
doi:10.1016/j.jim.2019.04.011 .

Nikolovski, Neda, Paunović, Verica, Stojanović, Ivana D., "Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells." in Journal of Immunological Methods, 470 (2019):46-54,
https://doi.org/10.1016/j.jim.2019.04.011 . .

TY  - JOUR
AU  - Stamenković, Marina
AU  - Janjetović, Kristina
AU  - Paunović, Verica
AU  - Ćirić, Darko
AU  - Kravić-Stevović, Tamara
AU  - Trajković, Vladimir
PY  - 2019
UR  - https://www.sciencedirect.com/science/article/pii/S0014299919304923?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3448
AB  - We performed a comparative analysis of molecular cytotoxic mechanisms of lysosomal autophagy inhibitors bafilomycin A1, chloroquine, and ammonium chloride in B16 mouse melanoma cells. All agents caused oxidative stress, mitochondrial depolarization, and caspase-dependent apoptotic death, which was not affected by genetic inactivation of autophagy. Cathepsin inhibition reduced only the cytotoxicity of chloroquine, indicating its ability to cause lysosomal membrane permeabilization. Bafilomycin reduced the mRNA levels of anti-apoptotic Bcl-2, while chloroquine and ammonium chloride increased the mRNA expression of pro-apoptotic Pten and Puma, as well as anti-apoptotic Bcl-xL. Ammonium chloride additionally increased the mRNA expression of pro-apoptotic Bim and p53. All three agents decreased the activity of mechanistic target of rapamycin (mTOR) and increased the activation of p38 mitogen-activated protein kinase (MAPK). Chloroquine and ammonium chloride additionally stimulated the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), respectively, while only bafilomycin increased the phosphorylation of the energy sensor AMP-activated protein kinase (AMPK). mTOR activator leucine did not affect the cytotoxicity of lysosomal inhibitors. p38 MAPK inhibitor SB203580 reduced the cytotoxicity of bafilomycin but increased that of chloroquine and ammonium chloride. The pharmacological inhibition of ERK1/2, JNK, and AMPK potentiated the cytotoxicity of chloroquine, ammonium chloride, and bafilomycin, respectively. The observed mechanistic differences were associated with antagonistic interactions of lysosomal inhibitors in B16 cell killing. In conclusion, all investigated lysosomal inhibitors cause autophagy-independent mitochondrial dysfunction and apoptotic death, but differ in the ability to affect lysosomal permeabilization, balance between pro- and anti-apoptotic molecules of Bcl-2 family, and MAPK/AMPK signaling.
T2  - European Journal of Pharmacology
T1  - Comparative analysis of cell death mechanisms induced by lysosomal autophagy inhibitors.
VL  - 859
DO  - 10.1016/j.ejphar.2019.172540
SP  - 172540
ER  - 

@article{
author = "Stamenković, Marina and Janjetović, Kristina and Paunović, Verica and Ćirić, Darko and Kravić-Stevović, Tamara and Trajković, Vladimir",
year = "2019",
abstract = "We performed a comparative analysis of molecular cytotoxic mechanisms of lysosomal autophagy inhibitors bafilomycin A1, chloroquine, and ammonium chloride in B16 mouse melanoma cells. All agents caused oxidative stress, mitochondrial depolarization, and caspase-dependent apoptotic death, which was not affected by genetic inactivation of autophagy. Cathepsin inhibition reduced only the cytotoxicity of chloroquine, indicating its ability to cause lysosomal membrane permeabilization. Bafilomycin reduced the mRNA levels of anti-apoptotic Bcl-2, while chloroquine and ammonium chloride increased the mRNA expression of pro-apoptotic Pten and Puma, as well as anti-apoptotic Bcl-xL. Ammonium chloride additionally increased the mRNA expression of pro-apoptotic Bim and p53. All three agents decreased the activity of mechanistic target of rapamycin (mTOR) and increased the activation of p38 mitogen-activated protein kinase (MAPK). Chloroquine and ammonium chloride additionally stimulated the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), respectively, while only bafilomycin increased the phosphorylation of the energy sensor AMP-activated protein kinase (AMPK). mTOR activator leucine did not affect the cytotoxicity of lysosomal inhibitors. p38 MAPK inhibitor SB203580 reduced the cytotoxicity of bafilomycin but increased that of chloroquine and ammonium chloride. The pharmacological inhibition of ERK1/2, JNK, and AMPK potentiated the cytotoxicity of chloroquine, ammonium chloride, and bafilomycin, respectively. The observed mechanistic differences were associated with antagonistic interactions of lysosomal inhibitors in B16 cell killing. In conclusion, all investigated lysosomal inhibitors cause autophagy-independent mitochondrial dysfunction and apoptotic death, but differ in the ability to affect lysosomal permeabilization, balance between pro- and anti-apoptotic molecules of Bcl-2 family, and MAPK/AMPK signaling.",
journal = "European Journal of Pharmacology",
title = "Comparative analysis of cell death mechanisms induced by lysosomal autophagy inhibitors.",
volume = "859",
doi = "10.1016/j.ejphar.2019.172540",
pages = "172540"
}

Stamenković, M., Janjetović, K., Paunović, V., Ćirić, D., Kravić-Stevović, T.,& Trajković, V.. (2019). Comparative analysis of cell death mechanisms induced by lysosomal autophagy inhibitors.. in European Journal of Pharmacology, 859, 172540.
https://doi.org/10.1016/j.ejphar.2019.172540

Stamenković M, Janjetović K, Paunović V, Ćirić D, Kravić-Stevović T, Trajković V. Comparative analysis of cell death mechanisms induced by lysosomal autophagy inhibitors.. in European Journal of Pharmacology. 2019;859:172540.
doi:10.1016/j.ejphar.2019.172540 .

Stamenković, Marina, Janjetović, Kristina, Paunović, Verica, Ćirić, Darko, Kravić-Stevović, Tamara, Trajković, Vladimir, "Comparative analysis of cell death mechanisms induced by lysosomal autophagy inhibitors." in European Journal of Pharmacology, 859 (2019):172540,
https://doi.org/10.1016/j.ejphar.2019.172540 . .