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

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

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

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

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

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

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

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

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

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