TY  - CONF
AU  - Đorđević, Marija
AU  - Feuerstein-Akgoz, Clarissa
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Sarić, Ana
AU  - Grdović, Nevena
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Mihailović, Mirjana
AU  - Gerhauser, Clarissa
AU  - Jurkowski, Tomasz
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6307
AB  - Introduction: The Aristaless-related homeobox (Arx) gene plays a key role in the development and maintaining pancreatic alpha cell phenotype, and as such represents an excellent target for alpha cell identity change towards insulin-producing cells. Therefore, this cell switch and increase in beta cell mass could be of potential use in diabetes management.
Methods: On the fifth day after transient transfection with dCas9-Dnmt3a3L-KRAB construct and four gRNAs targeting Arx promoter, αTC1-6 were sorted to collect GFP-positive (transfected) cells (EpiC). The mRNA-seq libraries were pooled in equimolar amounts and sequenced in a single end-setting on the Illumina NextSeq 500 High output machine with 75 bases long reads. KEGG pathway overrepresentation analysis was performed using an application on all significantly up- or downregulated genes using default settings.
Results: Directed induction of DNA methylation on the Arx gene promoter reduces its expression and causes the up-regulation of 357 genes, while 266 genes were down-regulated in EpiC compared to Mock transfected cells. The KEGG pathways analysis of biological processes confirmed several biological pathways associated with genes differentially expressed in EpiC vs. Mock transfected cells at the 5th posttransfection day (pval ≤ 0.05). As the most significant, up-regulated pathways we found Type II diabetes, Insulin secretion, Longevity regulation pathways. As significant, down-regulated pathways pop-up Fatty acid metabolism and PPAR signaling pathway.
Conclusion: Reduction of ArxmRNA level is sufficient to initiate the transdifferentiation process of alpha cells into insulin-producing cells by triggering several biological pathways tight related to insulin secretion and function.
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  - Changes in up and down regulated gene expression after transient suppression of Arx gene in pancreatic alphaTC1-6 cell line
SP  - 151
EP  - 151
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6307
ER  - 

@conference{
author = "Đorđević, Marija and Feuerstein-Akgoz, Clarissa and Arambašić Jovanović, Jelena and Tolić, Anja and Rajić, Jovana and Sarić, Ana and Grdović, Nevena and Dinić, Svetlana and Uskoković, Aleksandra and Mihailović, Mirjana and Gerhauser, Clarissa and Jurkowski, Tomasz and Vidaković, Melita",
year = "2023",
abstract = "Introduction: The Aristaless-related homeobox (Arx) gene plays a key role in the development and maintaining pancreatic alpha cell phenotype, and as such represents an excellent target for alpha cell identity change towards insulin-producing cells. Therefore, this cell switch and increase in beta cell mass could be of potential use in diabetes management.
Methods: On the fifth day after transient transfection with dCas9-Dnmt3a3L-KRAB construct and four gRNAs targeting Arx promoter, αTC1-6 were sorted to collect GFP-positive (transfected) cells (EpiC). The mRNA-seq libraries were pooled in equimolar amounts and sequenced in a single end-setting on the Illumina NextSeq 500 High output machine with 75 bases long reads. KEGG pathway overrepresentation analysis was performed using an application on all significantly up- or downregulated genes using default settings.
Results: Directed induction of DNA methylation on the Arx gene promoter reduces its expression and causes the up-regulation of 357 genes, while 266 genes were down-regulated in EpiC compared to Mock transfected cells. The KEGG pathways analysis of biological processes confirmed several biological pathways associated with genes differentially expressed in EpiC vs. Mock transfected cells at the 5th posttransfection day (pval ≤ 0.05). As the most significant, up-regulated pathways we found Type II diabetes, Insulin secretion, Longevity regulation pathways. As significant, down-regulated pathways pop-up Fatty acid metabolism and PPAR signaling pathway.
Conclusion: Reduction of ArxmRNA level is sufficient to initiate the transdifferentiation process of alpha cells into insulin-producing cells by triggering several biological pathways tight related to insulin secretion and function.",
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 = "Changes in up and down regulated gene expression after transient suppression of Arx gene in pancreatic alphaTC1-6 cell line",
pages = "151-151",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6307"
}

Đorđević, M., Feuerstein-Akgoz, C., Arambašić Jovanović, J., Tolić, A., Rajić, J., Sarić, A., Grdović, N., Dinić, S., Uskoković, A., Mihailović, M., Gerhauser, C., Jurkowski, T.,& Vidaković, M.. (2023). Changes in up and down regulated gene expression after transient suppression of Arx gene in pancreatic alphaTC1-6 cell line. 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., 151-151.
https://hdl.handle.net/21.15107/rcub_ibiss_6307

Đorđević M, Feuerstein-Akgoz C, Arambašić Jovanović J, Tolić A, Rajić J, Sarić A, Grdović N, Dinić S, Uskoković A, Mihailović M, Gerhauser C, Jurkowski T, Vidaković M. Changes in up and down regulated gene expression after transient suppression of Arx gene in pancreatic alphaTC1-6 cell line. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. 2023;:151-151.
https://hdl.handle.net/21.15107/rcub_ibiss_6307 .

Đorđević, Marija, Feuerstein-Akgoz, Clarissa, Arambašić Jovanović, Jelena, Tolić, Anja, Rajić, Jovana, Sarić, Ana, Grdović, Nevena, Dinić, Svetlana, Uskoković, Aleksandra, Mihailović, Mirjana, Gerhauser, Clarissa, Jurkowski, Tomasz, Vidaković, Melita, "Changes in up and down regulated gene expression after transient suppression of Arx gene in pancreatic alphaTC1-6 cell line" in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia (2023):151-151,
https://hdl.handle.net/21.15107/rcub_ibiss_6307 .

TY  - CONF
AU  - Sarić, Ana
AU  - Rajić, Jovana
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Grdović, Nevena
AU  - Uskoković, Aleksandra
AU  - Dinić, Svetlana
AU  - Arambašić Jovanović, Jelena
AU  - Mihailović, Mirjana
AU  - Vidaković, Melita
AU  - Dučić, Tanja
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6304
AB  - Introduction: DNA methylation is a major regulator of transcriptional activity and alongside other epigenetic modifications it introduces specific level of chromatin complexity. Fourier transform infrared (FTIR) spectroscopy is a rapid, non-destructive, and label-free technique for identifying subtle changes in all bio-macromolecules, and it has been used as a method of choice for studying DNA conformation. The present study was designed to explore the use of synchrotron-based FTIR spectroscopy to monitor the subtle changes on molecular level regarding the DNA methylation status of cytosine in the whole genome.
Methods: For FTIR-based DNA methylation analysis in situ, DNA-HALO samples were prepared using slightly modified methodology for nuclear HALO preparations where DNA-HALOs are liberated of any protein residues but preserve higher order chromatin structure.
Results: Using FTIR spectroscopy we analysed and compared methylation profiles of isolated genomic DNA and DNA-HALO samples. DNA-HALO structure shows more distinct peaks in fingerprint region of spectra. DNA-HALO structure is more accurate for detecting bonds in unmemthylated cytosine as specific infrared peaks are defined as vibrations of bonds in unmethylated cytosine at 1151 cm-1 and 1357 cm-1. The ratio of integrated area under the peak 1151 cm-1 over integrated area under the peak that represents PO2-backbone vibrations can be used to assess level of unmethylated cytosine and thus methylation rate in the DNA-HALO samples.
Conclusion: This study demonstrates potential of FTIR spectroscopy to detect DNA methylation in DNA-HALO samples more precisely compared to classical DNA extraction procedure that yield unstructured whole genomic DNA.
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  - Methylation profile analysis of Of DNA-HALO structure by synchrotron-based FTIR spectroscopy
SP  - 68
EP  - 68
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6304
ER  - 

@conference{
author = "Sarić, Ana and Rajić, Jovana and Tolić, Anja and Đorđević, Marija and Grdović, Nevena and Uskoković, Aleksandra and Dinić, Svetlana and Arambašić Jovanović, Jelena and Mihailović, Mirjana and Vidaković, Melita and Dučić, Tanja",
year = "2023",
abstract = "Introduction: DNA methylation is a major regulator of transcriptional activity and alongside other epigenetic modifications it introduces specific level of chromatin complexity. Fourier transform infrared (FTIR) spectroscopy is a rapid, non-destructive, and label-free technique for identifying subtle changes in all bio-macromolecules, and it has been used as a method of choice for studying DNA conformation. The present study was designed to explore the use of synchrotron-based FTIR spectroscopy to monitor the subtle changes on molecular level regarding the DNA methylation status of cytosine in the whole genome.
Methods: For FTIR-based DNA methylation analysis in situ, DNA-HALO samples were prepared using slightly modified methodology for nuclear HALO preparations where DNA-HALOs are liberated of any protein residues but preserve higher order chromatin structure.
Results: Using FTIR spectroscopy we analysed and compared methylation profiles of isolated genomic DNA and DNA-HALO samples. DNA-HALO structure shows more distinct peaks in fingerprint region of spectra. DNA-HALO structure is more accurate for detecting bonds in unmemthylated cytosine as specific infrared peaks are defined as vibrations of bonds in unmethylated cytosine at 1151 cm-1 and 1357 cm-1. The ratio of integrated area under the peak 1151 cm-1 over integrated area under the peak that represents PO2-backbone vibrations can be used to assess level of unmethylated cytosine and thus methylation rate in the DNA-HALO samples.
Conclusion: This study demonstrates potential of FTIR spectroscopy to detect DNA methylation in DNA-HALO samples more precisely compared to classical DNA extraction procedure that yield unstructured whole genomic DNA.",
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 = "Methylation profile analysis of Of DNA-HALO structure by synchrotron-based FTIR spectroscopy",
pages = "68-68",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6304"
}

Sarić, A., Rajić, J., Tolić, A., Đorđević, M., Grdović, N., Uskoković, A., Dinić, S., Arambašić Jovanović, J., Mihailović, M., Vidaković, M.,& Dučić, T.. (2023). Methylation profile analysis of Of DNA-HALO structure by synchrotron-based FTIR spectroscopy. 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., 68-68.
https://hdl.handle.net/21.15107/rcub_ibiss_6304

Sarić A, Rajić J, Tolić A, Đorđević M, Grdović N, Uskoković A, Dinić S, Arambašić Jovanović J, Mihailović M, Vidaković M, Dučić T. Methylation profile analysis of Of DNA-HALO structure by synchrotron-based FTIR spectroscopy. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. 2023;:68-68.
https://hdl.handle.net/21.15107/rcub_ibiss_6304 .

Sarić, Ana, Rajić, Jovana, Tolić, Anja, Đorđević, Marija, Grdović, Nevena, Uskoković, Aleksandra, Dinić, Svetlana, Arambašić Jovanović, Jelena, Mihailović, Mirjana, Vidaković, Melita, Dučić, Tanja, "Methylation profile analysis of Of DNA-HALO structure by synchrotron-based FTIR spectroscopy" in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia (2023):68-68,
https://hdl.handle.net/21.15107/rcub_ibiss_6304 .

TY  - JOUR
AU  - Sarić, Ana
AU  - Rajić, Jovana
AU  - Tolić, Anja
AU  - Dučić, Tanja
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5948
AB  - Fourier transform infrared (FTIR) spectroscopy is a rapid, non-destructive and label-free technique for identifying subtle changes in all bio-macromolecules, and has been used as a method of choice for studying DNA conformation, secondary DNA structure transition and DNA damage. In addition, the specific level of chromatin complexity is introduced via epigenetic modifications forcing the technological upgrade in the analysis of such an intricacy. As the most studied epigenetic mechanism, DNA methylation is a major regulator of transcriptional activity, involved in the suppression of a broad spectrum of genes and its deregulation is involved in all non-communicable diseases. The present study was designed to explore the use of synchrotron-based FTIR analysis to monitor the subtle changes in molecule bases regarding the DNA methylation status of cytosine in the whole genome. In order to reveal the conformation-related best sample for FTIR-based DNA methylation analysis in situ, we used methodology for nuclear HALO preparations and slightly modified it to isolated DNA in HALO formations. Nuclear DNA-HALOs represent samples with preserved higher-order chromatin structure liberated of any protein residues that are closer to native DNA conformation than genomic DNA (gDNA) isolated by the standard batch procedure. Using FTIR spectroscopy we analyzed the DNA methylation profile of isolated gDNA and compared it with the DNA-HALOs. This study demonstrated the potential of FTIR microspectroscopy to detect DNA methylation marks in analyzed DNA-HALO specimens more precisely in comparison with classical DNA extraction procedures that yield unstructured whole genomic DNA. In addition, we used different cell types to assess their global DNA methylation profile, as well as defined specific infrared peaks that can be used for screening DNA methylation.
PB  - Elsevier
T2  - Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy
T1  - Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure
IS  - 123090
VL  - 302
DO  - 10.1016/j.saa.2023.123090
ER  - 

@article{
author = "Sarić, Ana and Rajić, Jovana and Tolić, Anja and Dučić, Tanja and Vidaković, Melita",
year = "2023",
abstract = "Fourier transform infrared (FTIR) spectroscopy is a rapid, non-destructive and label-free technique for identifying subtle changes in all bio-macromolecules, and has been used as a method of choice for studying DNA conformation, secondary DNA structure transition and DNA damage. In addition, the specific level of chromatin complexity is introduced via epigenetic modifications forcing the technological upgrade in the analysis of such an intricacy. As the most studied epigenetic mechanism, DNA methylation is a major regulator of transcriptional activity, involved in the suppression of a broad spectrum of genes and its deregulation is involved in all non-communicable diseases. The present study was designed to explore the use of synchrotron-based FTIR analysis to monitor the subtle changes in molecule bases regarding the DNA methylation status of cytosine in the whole genome. In order to reveal the conformation-related best sample for FTIR-based DNA methylation analysis in situ, we used methodology for nuclear HALO preparations and slightly modified it to isolated DNA in HALO formations. Nuclear DNA-HALOs represent samples with preserved higher-order chromatin structure liberated of any protein residues that are closer to native DNA conformation than genomic DNA (gDNA) isolated by the standard batch procedure. Using FTIR spectroscopy we analyzed the DNA methylation profile of isolated gDNA and compared it with the DNA-HALOs. This study demonstrated the potential of FTIR microspectroscopy to detect DNA methylation marks in analyzed DNA-HALO specimens more precisely in comparison with classical DNA extraction procedures that yield unstructured whole genomic DNA. In addition, we used different cell types to assess their global DNA methylation profile, as well as defined specific infrared peaks that can be used for screening DNA methylation.",
publisher = "Elsevier",
journal = "Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy",
title = "Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure",
number = "123090",
volume = "302",
doi = "10.1016/j.saa.2023.123090"
}

Sarić, A., Rajić, J., Tolić, A., Dučić, T.,& Vidaković, M.. (2023). Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure. in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy
Elsevier., 302(123090).
https://doi.org/10.1016/j.saa.2023.123090

Sarić A, Rajić J, Tolić A, Dučić T, Vidaković M. Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure. in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2023;302(123090).
doi:10.1016/j.saa.2023.123090 .

Sarić, Ana, Rajić, Jovana, Tolić, Anja, Dučić, Tanja, Vidaković, Melita, "Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure" in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 302, no. 123090 (2023),
https://doi.org/10.1016/j.saa.2023.123090 . .

TY  - JOUR
AU  - Sarić, Ana
AU  - Rajić, Jovana
AU  - Tolić, Anja
AU  - Dučić, Tanja
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5947
AB  - Fourier transform infrared (FTIR) spectroscopy is a rapid, non-destructive and label-free technique for identifying subtle changes in all bio-macromolecules, and has been used as a method of choice for studying DNA conformation, secondary DNA structure transition and DNA damage. In addition, the specific level of chromatin complexity is introduced via epigenetic modifications forcing the technological upgrade in the analysis of such an intricacy. As the most studied epigenetic mechanism, DNA methylation is a major regulator of transcriptional activity, involved in the suppression of a broad spectrum of genes and its deregulation is involved in all non-communicable diseases. The present study was designed to explore the use of synchrotron-based FTIR analysis to monitor the subtle changes in molecule bases regarding the DNA methylation status of cytosine in the whole genome. In order to reveal the conformation-related best sample for FTIR-based DNA methylation analysis in situ, we used methodology for nuclear HALO preparations and slightly modified it to isolated DNA in HALO formations. Nuclear DNA-HALOs represent samples with preserved higher-order chromatin structure liberated of any protein residues that are closer to native DNA conformation than genomic DNA (gDNA) isolated by the standard batch procedure. Using FTIR spectroscopy we analyzed the DNA methylation profile of isolated gDNA and compared it with the DNA-HALOs. This study demonstrated the potential of FTIR microspectroscopy to detect DNA methylation marks in analyzed DNA-HALO specimens more precisely in comparison with classical DNA extraction procedures that yield unstructured whole genomic DNA. In addition, we used different cell types to assess their global DNA methylation profile, as well as defined specific infrared peaks that can be used for screening DNA methylation.
PB  - Elsevier
T2  - Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy
T1  - Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure
IS  - 123090
VL  - 302
DO  - 10.1016/j.saa.2023.123090
ER  - 

@article{
author = "Sarić, Ana and Rajić, Jovana and Tolić, Anja and Dučić, Tanja and Vidaković, Melita",
year = "2023",
abstract = "Fourier transform infrared (FTIR) spectroscopy is a rapid, non-destructive and label-free technique for identifying subtle changes in all bio-macromolecules, and has been used as a method of choice for studying DNA conformation, secondary DNA structure transition and DNA damage. In addition, the specific level of chromatin complexity is introduced via epigenetic modifications forcing the technological upgrade in the analysis of such an intricacy. As the most studied epigenetic mechanism, DNA methylation is a major regulator of transcriptional activity, involved in the suppression of a broad spectrum of genes and its deregulation is involved in all non-communicable diseases. The present study was designed to explore the use of synchrotron-based FTIR analysis to monitor the subtle changes in molecule bases regarding the DNA methylation status of cytosine in the whole genome. In order to reveal the conformation-related best sample for FTIR-based DNA methylation analysis in situ, we used methodology for nuclear HALO preparations and slightly modified it to isolated DNA in HALO formations. Nuclear DNA-HALOs represent samples with preserved higher-order chromatin structure liberated of any protein residues that are closer to native DNA conformation than genomic DNA (gDNA) isolated by the standard batch procedure. Using FTIR spectroscopy we analyzed the DNA methylation profile of isolated gDNA and compared it with the DNA-HALOs. This study demonstrated the potential of FTIR microspectroscopy to detect DNA methylation marks in analyzed DNA-HALO specimens more precisely in comparison with classical DNA extraction procedures that yield unstructured whole genomic DNA. In addition, we used different cell types to assess their global DNA methylation profile, as well as defined specific infrared peaks that can be used for screening DNA methylation.",
publisher = "Elsevier",
journal = "Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy",
title = "Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure",
number = "123090",
volume = "302",
doi = "10.1016/j.saa.2023.123090"
}

Sarić, A., Rajić, J., Tolić, A., Dučić, T.,& Vidaković, M.. (2023). Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure. in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy
Elsevier., 302(123090).
https://doi.org/10.1016/j.saa.2023.123090

Sarić A, Rajić J, Tolić A, Dučić T, Vidaković M. Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure. in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2023;302(123090).
doi:10.1016/j.saa.2023.123090 .

Sarić, Ana, Rajić, Jovana, Tolić, Anja, Dučić, Tanja, Vidaković, Melita, "Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure" in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 302, no. 123090 (2023),
https://doi.org/10.1016/j.saa.2023.123090 . .

TY  - DATA
AU  - Sarić, Ana
AU  - Rajić, Jovana
AU  - Tolić, Anja
AU  - Dučić, Tanja
AU  - Vidaković, Melita
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5960
AB  - Supplementary Figure 1:
Curve fitting of FTIR spectra for analyzing differences in DNA regions.
PB  - Elsevier
T2  - Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy
T1  - Supplementary material for the article: Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure
IS  - 123090
VL  - 302
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5960
ER  - 

@misc{
author = "Sarić, Ana and Rajić, Jovana and Tolić, Anja and Dučić, Tanja and Vidaković, Melita",
year = "2023",
abstract = "Supplementary Figure 1:
Curve fitting of FTIR spectra for analyzing differences in DNA regions.",
publisher = "Elsevier",
journal = "Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy",
title = "Supplementary material for the article: Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure",
number = "123090",
volume = "302",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5960"
}

Sarić, A., Rajić, J., Tolić, A., Dučić, T.,& Vidaković, M.. (2023). Supplementary material for the article: Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure. in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy
Elsevier., 302(123090).
https://hdl.handle.net/21.15107/rcub_ibiss_5960

Sarić A, Rajić J, Tolić A, Dučić T, Vidaković M. Supplementary material for the article: Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure. in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. 2023;302(123090).
https://hdl.handle.net/21.15107/rcub_ibiss_5960 .

Sarić, Ana, Rajić, Jovana, Tolić, Anja, Dučić, Tanja, Vidaković, Melita, "Supplementary material for the article: Synchrotron-based FTIR microspectroscopy reveals DNA methylation profile in DNA-HALO structure" in Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 302, no. 123090 (2023),
https://hdl.handle.net/21.15107/rcub_ibiss_5960 .

TY  - DATA
AU  - Jakubek, Patrycja
AU  - Rajić, Jovana
AU  - Kuczynska, Monika
AU  - Suliborska, Klaudia
AU  - Heldt, Mateusz
AU  - Dziedziul, Karol
AU  - Vidaković, Melita
AU  - Namiesnik, Jacek
AU  - Bartoszek, Agnieszka
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5670
AB  - Table S1. Primer sequences for SRXN1 gene used in MSP and MS-HRM. Table S2. Primer sequences for SRXN1 and GAPDH genes used in RT-qPCR. Table S3. Methylation changes in CpG island within the promoter region of SRXN1 in HT29 cell line investigated with MSP. Cells were treated with catechins and glutathione at concentrations of 0.1, 1,10, and 100 µM for 24 h at 37°C. The results are means ± SD of three biological replicates. Table S4. Methylation changes in CpG island within the promoter region of SRXN1 in HT29 cell line investigated with MS-HRM. Cells were treated with catechins and glutathione at concentrations of 0.1,1, 10, and 100 µM for 24 h at 37°C. The results are means ± SD of three biological replicates. Table S5. Changes in SRXN1 gene expression analysed with the RT-qPCR method. Expression levels were calculated using relative delta delta Ct method. Data was normalized to GAPDH gene expression. The results are means ± SD of three biological replicates. Figure S1. Inhibition of growth of HT29 cells determined by MTT assay after 72 h treatment with 5-Aza. Viability is expressed as a percentage relative to control cells (100%). EC30 and EC45 are the effective concentrations of 30% and 45% of cell growth inhibition. Results are means of three independent experiments carried out in triplicate (SD < 15%). Figure S2. The increase in DNA methylation level of SRXN1 gene determined by MS-HRM using M1/U1 primer sets following 24 h treatment of HT29 cells with catechins that increase methylation
profile of SRXN1 promoter. Figure S3. The increase in DNA methylation level of SRXN1 gene determined by MS-HRM using M3/U3 primer sets following 24 h treatment of HT29 cells with catechins that increase methylation profile of SRXN1 promoter.
PB  - Basel: MDPI
T2  - Antioxidants
T1  - Supplementary material for the article:Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene
IS  - 3
VL  - 12
SP  - 754
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5670
ER  - 

@misc{
author = "Jakubek, Patrycja and Rajić, Jovana and Kuczynska, Monika and Suliborska, Klaudia and Heldt, Mateusz and Dziedziul, Karol and Vidaković, Melita and Namiesnik, Jacek and Bartoszek, Agnieszka",
year = "2023",
abstract = "Table S1. Primer sequences for SRXN1 gene used in MSP and MS-HRM. Table S2. Primer sequences for SRXN1 and GAPDH genes used in RT-qPCR. Table S3. Methylation changes in CpG island within the promoter region of SRXN1 in HT29 cell line investigated with MSP. Cells were treated with catechins and glutathione at concentrations of 0.1, 1,10, and 100 µM for 24 h at 37°C. The results are means ± SD of three biological replicates. Table S4. Methylation changes in CpG island within the promoter region of SRXN1 in HT29 cell line investigated with MS-HRM. Cells were treated with catechins and glutathione at concentrations of 0.1,1, 10, and 100 µM for 24 h at 37°C. The results are means ± SD of three biological replicates. Table S5. Changes in SRXN1 gene expression analysed with the RT-qPCR method. Expression levels were calculated using relative delta delta Ct method. Data was normalized to GAPDH gene expression. The results are means ± SD of three biological replicates. Figure S1. Inhibition of growth of HT29 cells determined by MTT assay after 72 h treatment with 5-Aza. Viability is expressed as a percentage relative to control cells (100%). EC30 and EC45 are the effective concentrations of 30% and 45% of cell growth inhibition. Results are means of three independent experiments carried out in triplicate (SD < 15%). Figure S2. The increase in DNA methylation level of SRXN1 gene determined by MS-HRM using M1/U1 primer sets following 24 h treatment of HT29 cells with catechins that increase methylation
profile of SRXN1 promoter. Figure S3. The increase in DNA methylation level of SRXN1 gene determined by MS-HRM using M3/U3 primer sets following 24 h treatment of HT29 cells with catechins that increase methylation profile of SRXN1 promoter.",
publisher = "Basel: MDPI",
journal = "Antioxidants",
title = "Supplementary material for the article:Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene",
number = "3",
volume = "12",
pages = "754",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5670"
}

Jakubek, P., Rajić, J., Kuczynska, M., Suliborska, K., Heldt, M., Dziedziul, K., Vidaković, M., Namiesnik, J.,& Bartoszek, A.. (2023). Supplementary material for the article:Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene. in Antioxidants
Basel: MDPI., 12(3), 754.
https://hdl.handle.net/21.15107/rcub_ibiss_5670

Jakubek P, Rajić J, Kuczynska M, Suliborska K, Heldt M, Dziedziul K, Vidaković M, Namiesnik J, Bartoszek A. Supplementary material for the article:Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene. in Antioxidants. 2023;12(3):754.
https://hdl.handle.net/21.15107/rcub_ibiss_5670 .

Jakubek, Patrycja, Rajić, Jovana, Kuczynska, Monika, Suliborska, Klaudia, Heldt, Mateusz, Dziedziul, Karol, Vidaković, Melita, Namiesnik, Jacek, Bartoszek, Agnieszka, "Supplementary material for the article:Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene" in Antioxidants, 12, no. 3 (2023):754,
https://hdl.handle.net/21.15107/rcub_ibiss_5670 .

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  - Jakubek, Patrycja
AU  - Rajić, Jovana
AU  - Kuczynska, Monika
AU  - Suliborska, Klaudia
AU  - Heldt, Mateusz
AU  - Dziedziul, Karol
AU  - Vidaković, Melita
AU  - Namiesnik, Jacek
AU  - Bartoszek, Agnieszka
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5506
AB  - The role of catechins in the epigenetic regulation of gene expression has been widely studied; however, if and how this phenomenon relates to the redox properties of these polyphenols remains unknown. Our earlier study demonstrated that exposure of the human colon adenocarcinoma HT29 cell line to these antioxidants affects the expression of redox-related genes. In particular, treatment with (−)-epigallocatechin (EGC) downregulated transcription of gene encoding sulfiredoxin-1 (SRXN1), the peroxidase involved in the protection of cells against hydrogen peroxide-induced oxidative stress. The aim of this study was to investigate whether the observed SRXN1 downregulation was accompanied by changes in the DNA methylation level of its promoter and, if so, whether it was correlated with the redox properties of catechins. The impact on DNA methylation profile in HT29 cells treated with different concentrations of five catechins, varying in chemical structures and standard reduction potentials as well as susceptibility to oxidation, was monitored by a methylation-sensitive high-resolution melting technique employing the SRXN1 promoter region as a model target. We demonstrated that catechins, indeed, are able to modulate DNA methylation of the SRXN1 gene in a redox-related manner. The nonlinear method in the statistical analysis made it possible to fish out two parameters (charge transfer in oxidation process Qox and time of electron transfer t), whose strong interactions correlated with observed modulation of DNA methylation by catechins. Based on these findings, we present a proof-of-concept that DNA methylation, which limits SRXN1 expression and thus restricts the multidirectional antioxidant action of SRXN1, may represent a mechanism protecting cells against reductive stress caused by particularly fast-reacting reductants such as EGC and (−)-epicatechin gallate (ECG) in our study.
PB  - Basel: MDPI
T2  - Antioxidants
T1  - Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene
IS  - 3
VL  - 12
DO  - 10.3390/antiox12030754
SP  - 754
ER  - 

@article{
author = "Jakubek, Patrycja and Rajić, Jovana and Kuczynska, Monika and Suliborska, Klaudia and Heldt, Mateusz and Dziedziul, Karol and Vidaković, Melita and Namiesnik, Jacek and Bartoszek, Agnieszka",
year = "2023",
abstract = "The role of catechins in the epigenetic regulation of gene expression has been widely studied; however, if and how this phenomenon relates to the redox properties of these polyphenols remains unknown. Our earlier study demonstrated that exposure of the human colon adenocarcinoma HT29 cell line to these antioxidants affects the expression of redox-related genes. In particular, treatment with (−)-epigallocatechin (EGC) downregulated transcription of gene encoding sulfiredoxin-1 (SRXN1), the peroxidase involved in the protection of cells against hydrogen peroxide-induced oxidative stress. The aim of this study was to investigate whether the observed SRXN1 downregulation was accompanied by changes in the DNA methylation level of its promoter and, if so, whether it was correlated with the redox properties of catechins. The impact on DNA methylation profile in HT29 cells treated with different concentrations of five catechins, varying in chemical structures and standard reduction potentials as well as susceptibility to oxidation, was monitored by a methylation-sensitive high-resolution melting technique employing the SRXN1 promoter region as a model target. We demonstrated that catechins, indeed, are able to modulate DNA methylation of the SRXN1 gene in a redox-related manner. The nonlinear method in the statistical analysis made it possible to fish out two parameters (charge transfer in oxidation process Qox and time of electron transfer t), whose strong interactions correlated with observed modulation of DNA methylation by catechins. Based on these findings, we present a proof-of-concept that DNA methylation, which limits SRXN1 expression and thus restricts the multidirectional antioxidant action of SRXN1, may represent a mechanism protecting cells against reductive stress caused by particularly fast-reacting reductants such as EGC and (−)-epicatechin gallate (ECG) in our study.",
publisher = "Basel: MDPI",
journal = "Antioxidants",
title = "Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene",
number = "3",
volume = "12",
doi = "10.3390/antiox12030754",
pages = "754"
}

Jakubek, P., Rajić, J., Kuczynska, M., Suliborska, K., Heldt, M., Dziedziul, K., Vidaković, M., Namiesnik, J.,& Bartoszek, A.. (2023). Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene. in Antioxidants
Basel: MDPI., 12(3), 754.
https://doi.org/10.3390/antiox12030754

Jakubek P, Rajić J, Kuczynska M, Suliborska K, Heldt M, Dziedziul K, Vidaković M, Namiesnik J, Bartoszek A. Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene. in Antioxidants. 2023;12(3):754.
doi:10.3390/antiox12030754 .

Jakubek, Patrycja, Rajić, Jovana, Kuczynska, Monika, Suliborska, Klaudia, Heldt, Mateusz, Dziedziul, Karol, Vidaković, Melita, Namiesnik, Jacek, Bartoszek, Agnieszka, "Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile—The Example of SRXN1 Gene" in Antioxidants, 12, no. 3 (2023):754,
https://doi.org/10.3390/antiox12030754 . .

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  - JOUR
AU  - Dinić, Svetlana
AU  - Arambašić Jovanović, Jelena
AU  - Uskoković, Aleksandra
AU  - Mihailović, Mirjana
AU  - Grdović, Nevena
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Đorđević, Marija
AU  - Vidaković, Melita
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5125
AB  - The biggest drawback of a current diabetes therapy is the treatment of the consequences not the cause of the disease. Regardless of the diabetes type, preservation and recovery of functional pancreatic beta cells stands as the biggest challenge in the treatment of diabetes. Free radicals and oxidative stress are among the major mediators of autoimmune destruction of beta cells in type 1 diabetes (T1D) or beta cell malfunction and death provoked by glucotoxicity and insulin resistance in type 2 diabetes (T2D). Additionally, oxidative stress reduces functionality of beta cells in T2D by stimulating their de-/trans-differentiation through the loss of transcription factors critical for beta cell development, maturity and regeneration. This review summarizes up to date clarified redox-related mechanisms involved in regulating beta cell identity and death, underlining similarities and differences between T1D and T2D. The protective effects of natural antioxidants on the oxidative stress-induced beta cell failure were also discussed. Considering that oxidative stress affects epigenetic regulatory mechanisms involved in the regulation of pancreatic beta cell survival and insulin secretion, this review highlighted huge potential of epigenetic therapy. Special attention was paid on application of the state-of-the-art CRISPR/Cas9 technology, based on targeted epigenome editing with the purpose of changing the differentiation state of different cell types, making them insulin-producing with ability to attenuate diabetes. Clarification of the above-mentioned mechanisms could provide better insight into diabetes etiology and pathogenesis, which would allow development of novel, potentially more efficient therapeutic strategies for the prevention or reversion of beta cell loss.
PB  - Lausanne: Frontiers Media SA
T2  - Frontiers in Endocrinology
T1  - Oxidative stress-mediated beta cell death and dysfunction as a target for diabetes management
VL  - 13
DO  - 10.3389/fendo.2022.1006376
SP  - 1006376
ER  - 

@article{
author = "Dinić, Svetlana and Arambašić Jovanović, Jelena and Uskoković, Aleksandra and Mihailović, Mirjana and Grdović, Nevena and Tolić, Anja and Rajić, Jovana and Đorđević, Marija and Vidaković, Melita",
year = "2022",
abstract = "The biggest drawback of a current diabetes therapy is the treatment of the consequences not the cause of the disease. Regardless of the diabetes type, preservation and recovery of functional pancreatic beta cells stands as the biggest challenge in the treatment of diabetes. Free radicals and oxidative stress are among the major mediators of autoimmune destruction of beta cells in type 1 diabetes (T1D) or beta cell malfunction and death provoked by glucotoxicity and insulin resistance in type 2 diabetes (T2D). Additionally, oxidative stress reduces functionality of beta cells in T2D by stimulating their de-/trans-differentiation through the loss of transcription factors critical for beta cell development, maturity and regeneration. This review summarizes up to date clarified redox-related mechanisms involved in regulating beta cell identity and death, underlining similarities and differences between T1D and T2D. The protective effects of natural antioxidants on the oxidative stress-induced beta cell failure were also discussed. Considering that oxidative stress affects epigenetic regulatory mechanisms involved in the regulation of pancreatic beta cell survival and insulin secretion, this review highlighted huge potential of epigenetic therapy. Special attention was paid on application of the state-of-the-art CRISPR/Cas9 technology, based on targeted epigenome editing with the purpose of changing the differentiation state of different cell types, making them insulin-producing with ability to attenuate diabetes. Clarification of the above-mentioned mechanisms could provide better insight into diabetes etiology and pathogenesis, which would allow development of novel, potentially more efficient therapeutic strategies for the prevention or reversion of beta cell loss.",
publisher = "Lausanne: Frontiers Media SA",
journal = "Frontiers in Endocrinology",
title = "Oxidative stress-mediated beta cell death and dysfunction as a target for diabetes management",
volume = "13",
doi = "10.3389/fendo.2022.1006376",
pages = "1006376"
}

Dinić, S., Arambašić Jovanović, J., Uskoković, A., Mihailović, M., Grdović, N., Tolić, A., Rajić, J., Đorđević, M.,& Vidaković, M.. (2022). Oxidative stress-mediated beta cell death and dysfunction as a target for diabetes management. in Frontiers in Endocrinology
Lausanne: Frontiers Media SA., 13, 1006376.
https://doi.org/10.3389/fendo.2022.1006376

Dinić S, Arambašić Jovanović J, Uskoković A, Mihailović M, Grdović N, Tolić A, Rajić J, Đorđević M, Vidaković M. Oxidative stress-mediated beta cell death and dysfunction as a target for diabetes management. in Frontiers in Endocrinology. 2022;13:1006376.
doi:10.3389/fendo.2022.1006376 .

Dinić, Svetlana, Arambašić Jovanović, Jelena, Uskoković, Aleksandra, Mihailović, Mirjana, Grdović, Nevena, Tolić, Anja, Rajić, Jovana, Đorđević, Marija, Vidaković, Melita, "Oxidative stress-mediated beta cell death and dysfunction as a target for diabetes management" in Frontiers in Endocrinology, 13 (2022):1006376,
https://doi.org/10.3389/fendo.2022.1006376 . .

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  - JOUR
AU  - Tolić, Anja
AU  - Ravichandran, Mirunalini
AU  - Rajić, Jovana
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Dinić, Svetlana
AU  - Grdović, Nevena
AU  - Arambašić Jovanović, Jelena
AU  - Mihailović, Mirjana
AU  - Nestorović, Nataša
AU  - Jurkowski, Tomasz P.
AU  - Uskoković, Aleksandra
AU  - Vidaković, Melita
PY  - 2022
UR  - https://epigeneticsandchromatin.biomedcentral.com/articles/10.1186/s13072-022-00445-8
UR  - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=PMC8985375
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4957
AB  - BACKGROUND Poly(ADP-ribosyl)ation (PARylation), a posttranslational modification introduced by PARP-1 and PARP-2, has first been implicated in DNA demethylation due to its role in base excision repair. Recent evidence indicates a direct influence of PARP-dependent PARylation on TET enzymes which catalyse hydroxymethylation of DNA-the first step in DNA demethylation. However, the exact nature of influence that PARylation exerts on TET activity is still ambiguous. In our recent study, we have observed a negative influence of PARP-1 on local TET-mediated DNA demethylation of a single gene and in this study, we further explore PARP-TET interplay. RESULTS Expanding on our previous work, we show that both TET1 and TET2 can be in vitro PARylated by PARP-1 and PARP-2 enzymes and that TET1 PARylation negatively affects the TET1 catalytic activity in vitro. Furthermore, we show that PARylation inhibits TET-mediated DNA demethylation at the global genome level in cellulo. CONCLUSIONS According to our findings, PARP inhibition can positively influence TET activity and therefore affect global levels of DNA methylation and hydroxymethylation. This gives a strong rationale for future examination of PARP inhibitors' potential use in the therapy of cancers characterised by loss of 5-hydroxymethylcytosine.
PB  - London: BioMed Central Ltd
T2  - Epigenetics & Chromatin
T1  - TET-mediated DNA hydroxymethylation is negatively influenced by the PARP-dependent PARylation.
IS  - 1
VL  - 15
DO  - 10.1186/s13072-022-00445-8
SP  - 11
ER  - 

@article{
author = "Tolić, Anja and Ravichandran, Mirunalini and Rajić, Jovana and Đorđević, Marija and Đorđević, Miloš and Dinić, Svetlana and Grdović, Nevena and Arambašić Jovanović, Jelena and Mihailović, Mirjana and Nestorović, Nataša and Jurkowski, Tomasz P. and Uskoković, Aleksandra and Vidaković, Melita",
year = "2022",
abstract = "BACKGROUND Poly(ADP-ribosyl)ation (PARylation), a posttranslational modification introduced by PARP-1 and PARP-2, has first been implicated in DNA demethylation due to its role in base excision repair. Recent evidence indicates a direct influence of PARP-dependent PARylation on TET enzymes which catalyse hydroxymethylation of DNA-the first step in DNA demethylation. However, the exact nature of influence that PARylation exerts on TET activity is still ambiguous. In our recent study, we have observed a negative influence of PARP-1 on local TET-mediated DNA demethylation of a single gene and in this study, we further explore PARP-TET interplay. RESULTS Expanding on our previous work, we show that both TET1 and TET2 can be in vitro PARylated by PARP-1 and PARP-2 enzymes and that TET1 PARylation negatively affects the TET1 catalytic activity in vitro. Furthermore, we show that PARylation inhibits TET-mediated DNA demethylation at the global genome level in cellulo. CONCLUSIONS According to our findings, PARP inhibition can positively influence TET activity and therefore affect global levels of DNA methylation and hydroxymethylation. This gives a strong rationale for future examination of PARP inhibitors' potential use in the therapy of cancers characterised by loss of 5-hydroxymethylcytosine.",
publisher = "London: BioMed Central Ltd",
journal = "Epigenetics & Chromatin",
title = "TET-mediated DNA hydroxymethylation is negatively influenced by the PARP-dependent PARylation.",
number = "1",
volume = "15",
doi = "10.1186/s13072-022-00445-8",
pages = "11"
}

Tolić, A., Ravichandran, M., Rajić, J., Đorđević, M., Đorđević, M., Dinić, S., Grdović, N., Arambašić Jovanović, J., Mihailović, M., Nestorović, N., Jurkowski, T. P., Uskoković, A.,& Vidaković, M.. (2022). TET-mediated DNA hydroxymethylation is negatively influenced by the PARP-dependent PARylation.. in Epigenetics & Chromatin
London: BioMed Central Ltd., 15(1), 11.
https://doi.org/10.1186/s13072-022-00445-8

Tolić A, Ravichandran M, Rajić J, Đorđević M, Đorđević M, Dinić S, Grdović N, Arambašić Jovanović J, Mihailović M, Nestorović N, Jurkowski TP, Uskoković A, Vidaković M. TET-mediated DNA hydroxymethylation is negatively influenced by the PARP-dependent PARylation.. in Epigenetics & Chromatin. 2022;15(1):11.
doi:10.1186/s13072-022-00445-8 .

Tolić, Anja, Ravichandran, Mirunalini, Rajić, Jovana, Đorđević, Marija, Đorđević, Miloš, Dinić, Svetlana, Grdović, Nevena, Arambašić Jovanović, Jelena, Mihailović, Mirjana, Nestorović, Nataša, Jurkowski, Tomasz P., Uskoković, Aleksandra, Vidaković, Melita, "TET-mediated DNA hydroxymethylation is negatively influenced by the PARP-dependent PARylation." in Epigenetics & Chromatin, 15, no. 1 (2022):11,
https://doi.org/10.1186/s13072-022-00445-8 . .

TY  - JOUR
AU  - Đorđević, Miloš
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Uskoković, Aleksandra
AU  - Grdović, Nevena
AU  - Đorđević, Marija
AU  - Mišić, Danijela
AU  - Šiler, Branislav
AU  - Vidaković, Melita
AU  - Dinić, Svetlana
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4782
AB  - The use of medicinal herbs can mitigate oxidative stress-induced diabetic complications and organ failure. This study investigated hepato- and reno-protective effects of methanol extract of Centaurium erythraea Rafn (CEE) in STZ-diabetic rats pre-treated (2 weeks) and/or post-treated (4 weeks) with CEE (orally, 100 mg/kg/day). Both CEE treatments markedly improved liver and kidney functionality in diabetic rats observed as reduced aspartate and alanine aminotransferase activities and decreased creatinine and blood urea nitrogen levels. CEE pre-treatment reduced the level of glycosylated proteins in diabetic liver more efficiently than post-treatment. Lowered levels of lipid peroxidation, DNA damage and protein glutathionylation, elevated ratio of reduced to oxidized glutathione, and mitigated disturbance of antioxidant enzyme activities reflected the antioxidant effect of CEE in diabetic liver and kidney. Although CEE pre-treatment was more effective, the obtained results indicate that both treatments protect the liver and kidney from oxidative damage by boosting the endogenous antioxidant defense system.
PB  - Elsevier
T2  - Journal of Functional Foods
T1  - Centaurium erythraea methanol extract improves the functionality of diabetic liver and kidney by mitigating hyperglycemia-induced oxidative stress
VL  - 90
DO  - 10.1016/j.jff.2022.104975
SP  - 104975
ER  - 

@article{
author = "Đorđević, Miloš and Tolić, Anja and Rajić, Jovana and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Uskoković, Aleksandra and Grdović, Nevena and Đorđević, Marija and Mišić, Danijela and Šiler, Branislav and Vidaković, Melita and Dinić, Svetlana",
year = "2022",
abstract = "The use of medicinal herbs can mitigate oxidative stress-induced diabetic complications and organ failure. This study investigated hepato- and reno-protective effects of methanol extract of Centaurium erythraea Rafn (CEE) in STZ-diabetic rats pre-treated (2 weeks) and/or post-treated (4 weeks) with CEE (orally, 100 mg/kg/day). Both CEE treatments markedly improved liver and kidney functionality in diabetic rats observed as reduced aspartate and alanine aminotransferase activities and decreased creatinine and blood urea nitrogen levels. CEE pre-treatment reduced the level of glycosylated proteins in diabetic liver more efficiently than post-treatment. Lowered levels of lipid peroxidation, DNA damage and protein glutathionylation, elevated ratio of reduced to oxidized glutathione, and mitigated disturbance of antioxidant enzyme activities reflected the antioxidant effect of CEE in diabetic liver and kidney. Although CEE pre-treatment was more effective, the obtained results indicate that both treatments protect the liver and kidney from oxidative damage by boosting the endogenous antioxidant defense system.",
publisher = "Elsevier",
journal = "Journal of Functional Foods",
title = "Centaurium erythraea methanol extract improves the functionality of diabetic liver and kidney by mitigating hyperglycemia-induced oxidative stress",
volume = "90",
doi = "10.1016/j.jff.2022.104975",
pages = "104975"
}

Đorđević, M., Tolić, A., Rajić, J., Mihailović, M., Arambašić Jovanović, J., Uskoković, A., Grdović, N., Đorđević, M., Mišić, D., Šiler, B., Vidaković, M.,& Dinić, S.. (2022). Centaurium erythraea methanol extract improves the functionality of diabetic liver and kidney by mitigating hyperglycemia-induced oxidative stress. in Journal of Functional Foods
Elsevier., 90, 104975.
https://doi.org/10.1016/j.jff.2022.104975

Đorđević M, Tolić A, Rajić J, Mihailović M, Arambašić Jovanović J, Uskoković A, Grdović N, Đorđević M, Mišić D, Šiler B, Vidaković M, Dinić S. Centaurium erythraea methanol extract improves the functionality of diabetic liver and kidney by mitigating hyperglycemia-induced oxidative stress. in Journal of Functional Foods. 2022;90:104975.
doi:10.1016/j.jff.2022.104975 .

Đorđević, Miloš, Tolić, Anja, Rajić, Jovana, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Uskoković, Aleksandra, Grdović, Nevena, Đorđević, Marija, Mišić, Danijela, Šiler, Branislav, Vidaković, Melita, Dinić, Svetlana, "Centaurium erythraea methanol extract improves the functionality of diabetic liver and kidney by mitigating hyperglycemia-induced oxidative stress" in Journal of Functional Foods, 90 (2022):104975,
https://doi.org/10.1016/j.jff.2022.104975 . .

TY  - CONF
AU  - Sarić, Ana
AU  - Rajić, Jovana
AU  - Đorđević, Marija
AU  - Jovanović, Mirna
AU  - Podolski-Renić, Ana
AU  - Pešić, Milica
AU  - Uskoković, Aleksandra
AU  - Tolić, Anja
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5918
AB  - 5-hydroxymethylcytosine (5-hmC) is formed through the oxidation of 5-methylcytosine (5-mC) by the catalytic activity of TET enzymes. It has been proposed to have regulatory functions as an independent epigenetic mark and it might have diagnostic potential as global loss of 5hmC level is commonly observed in cancers.
We have recently shown the inhibitory influence of PARP-1 dependent PARylation on TET1 hydroxylase activity in DNA demethylation. These findings could provide the rationale for using PARP inhibitors in cancers that are characterised by the 5hmC loss, other than cancer where treatment with PARP inhibitors exploited homologues repair (HR) defects on the basis of the synthetic lethality phenomenon. The activating effects of PARP inhibition on TET activity could provide the additional mechanism of action of PARP inhibitors (which are less cytotoxic than standard chemotherapeutic agents) in the treatment of cancers characterised by diminishing levels of 5hmC.
In our study, we have tested the effects of PARP inhibition on 5-hmC levels in non-small cell lung cancer. First, we analysed global levels of 5-hmC in several non-small cell lung cancer cell lines (NCI-H460 sensitive and resistant to doxorubicin, A549, NCI-H661) in comparison to normal fetal lung fibroblast cells MRC-5. Since both slot-blot and confocal microscopy analyses have shown that the A549 cell line has the lowest level of 5-hmC this cell line was selected for further experiments in which we set out to raise DNA hydroxymethylation levels by inhibiting PARP activity. After 72h treatment of A549 cells with PARylation inhibitor niraparib (IC50= 10 μM), we indeed observed an increase in 5-hmC level while 5-mC level did not change. This is a promising start to our investigation of alternative mechanisms of action of PARP inhibition in the treatment of cancers. Through this research, we hope to expand the range of cancer types that would be treated with PARP inhibitors, regardless of the HR status.
PB  - Josep Carreras Leukaemia Research Institute (IJC)
PB  - Innovative Training Network (ITN) INTERCEPT-MDS
C3  - Spetses summer school: Cancer epigenetics: Principles, Applications and Single-cell resolution; 2022 Aug 28 - Sep 3; Spteses Island, Greece
T1  - Influence of PARP inhibition on 5-hmC level in NSCLC
SP  - 116
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5918
ER  - 

@conference{
author = "Sarić, Ana and Rajić, Jovana and Đorđević, Marija and Jovanović, Mirna and Podolski-Renić, Ana and Pešić, Milica and Uskoković, Aleksandra and Tolić, Anja",
year = "2022",
abstract = "5-hydroxymethylcytosine (5-hmC) is formed through the oxidation of 5-methylcytosine (5-mC) by the catalytic activity of TET enzymes. It has been proposed to have regulatory functions as an independent epigenetic mark and it might have diagnostic potential as global loss of 5hmC level is commonly observed in cancers.
We have recently shown the inhibitory influence of PARP-1 dependent PARylation on TET1 hydroxylase activity in DNA demethylation. These findings could provide the rationale for using PARP inhibitors in cancers that are characterised by the 5hmC loss, other than cancer where treatment with PARP inhibitors exploited homologues repair (HR) defects on the basis of the synthetic lethality phenomenon. The activating effects of PARP inhibition on TET activity could provide the additional mechanism of action of PARP inhibitors (which are less cytotoxic than standard chemotherapeutic agents) in the treatment of cancers characterised by diminishing levels of 5hmC.
In our study, we have tested the effects of PARP inhibition on 5-hmC levels in non-small cell lung cancer. First, we analysed global levels of 5-hmC in several non-small cell lung cancer cell lines (NCI-H460 sensitive and resistant to doxorubicin, A549, NCI-H661) in comparison to normal fetal lung fibroblast cells MRC-5. Since both slot-blot and confocal microscopy analyses have shown that the A549 cell line has the lowest level of 5-hmC this cell line was selected for further experiments in which we set out to raise DNA hydroxymethylation levels by inhibiting PARP activity. After 72h treatment of A549 cells with PARylation inhibitor niraparib (IC50= 10 μM), we indeed observed an increase in 5-hmC level while 5-mC level did not change. This is a promising start to our investigation of alternative mechanisms of action of PARP inhibition in the treatment of cancers. Through this research, we hope to expand the range of cancer types that would be treated with PARP inhibitors, regardless of the HR status.",
publisher = "Josep Carreras Leukaemia Research Institute (IJC), Innovative Training Network (ITN) INTERCEPT-MDS",
journal = "Spetses summer school: Cancer epigenetics: Principles, Applications and Single-cell resolution; 2022 Aug 28 - Sep 3; Spteses Island, Greece",
title = "Influence of PARP inhibition on 5-hmC level in NSCLC",
pages = "116",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5918"
}

Sarić, A., Rajić, J., Đorđević, M., Jovanović, M., Podolski-Renić, A., Pešić, M., Uskoković, A.,& Tolić, A.. (2022). Influence of PARP inhibition on 5-hmC level in NSCLC. in Spetses summer school: Cancer epigenetics: Principles, Applications and Single-cell resolution; 2022 Aug 28 - Sep 3; Spteses Island, Greece
Josep Carreras Leukaemia Research Institute (IJC)., 116.
https://hdl.handle.net/21.15107/rcub_ibiss_5918

Sarić A, Rajić J, Đorđević M, Jovanović M, Podolski-Renić A, Pešić M, Uskoković A, Tolić A. Influence of PARP inhibition on 5-hmC level in NSCLC. in Spetses summer school: Cancer epigenetics: Principles, Applications and Single-cell resolution; 2022 Aug 28 - Sep 3; Spteses Island, Greece. 2022;:116.
https://hdl.handle.net/21.15107/rcub_ibiss_5918 .

Sarić, Ana, Rajić, Jovana, Đorđević, Marija, Jovanović, Mirna, Podolski-Renić, Ana, Pešić, Milica, Uskoković, Aleksandra, Tolić, Anja, "Influence of PARP inhibition on 5-hmC level in NSCLC" in Spetses summer school: Cancer epigenetics: Principles, Applications and Single-cell resolution; 2022 Aug 28 - Sep 3; Spteses Island, Greece (2022):116,
https://hdl.handle.net/21.15107/rcub_ibiss_5918 .

TY  - JOUR
AU  - Grdović, Nevena
AU  - Rajić, Jovana
AU  - Arambašić Jovanović, Jelena
AU  - Dinić, Svetlana
AU  - Tolić, Anja
AU  - Đorđević, Miloš
AU  - Đorđević, Marija
AU  - Trifunović, Svetlana
AU  - Vidaković, Melita
AU  - Uskoković, Aleksandra
AU  - Mihailović, Mirjana
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4190
AB  - α-Lipoic acid (ALA) is widely used as a nutritional supplement and therapeutic agent in diabetes management. Well-established antioxidant and hypoglycemic effects of ALA were considered to be particularly important in combating diabetic complications including renal injury. The present study evaluated the potential of ALA to affect profibrotic events in kidney that could alter its structure and functioning. ALA was administered intraperitoneally (10 mg/kg) to nondiabetic and streptozotocin-induced diabetic male Wistar rats for 4 and 8 weeks. The effects of ALA were assessed starting from structural/morphological alterations through changes that characterize profibrotic processes, to regulation of collagen gene expression in kidney. Here, we demonstrated that ALA improved systemic glucose and urea level, reduced formation of renal advanced glycation end products (AGEs), and maintained renal structural integrity in diabetic rats. However, profibrotic events provoked in diabetes were not alleviated by ALA since collagen synthesis/deposition and expression of transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) remained elevated in ALA-treated diabetic rats, especially after 8 weeks of diabetes onset. Moreover, 8 weeks treatment of nondiabetic rats with ALA led to the development of profibrotic features reflected in increased collagen synthesis/deposition. Besides the TGF-β1 downstream signaling, the additional mechanism underlying the upregulation of collagen IV in nondiabetic rats treated with ALA involves decreased DNA methylation of its promoter that could arise from increased Tet1 expression. These findings emphasize the therapeutic caution in the use of ALA, especially in patients with renal diabetic complication.
PB  - Hindawi Limited
T2  - Oxidative Medicine and Cellular Longevity
T1  - α-Lipoic Acid Increases Collagen Synthesis and Deposition in Nondiabetic and Diabetic Rat Kidneys
VL  - 2021
DO  - 10.1155/2021/6669352
SP  - 6669352
ER  - 

@article{
author = "Grdović, Nevena and Rajić, Jovana and Arambašić Jovanović, Jelena and Dinić, Svetlana and Tolić, Anja and Đorđević, Miloš and Đorđević, Marija and Trifunović, Svetlana and Vidaković, Melita and Uskoković, Aleksandra and Mihailović, Mirjana",
year = "2021",
abstract = "α-Lipoic acid (ALA) is widely used as a nutritional supplement and therapeutic agent in diabetes management. Well-established antioxidant and hypoglycemic effects of ALA were considered to be particularly important in combating diabetic complications including renal injury. The present study evaluated the potential of ALA to affect profibrotic events in kidney that could alter its structure and functioning. ALA was administered intraperitoneally (10 mg/kg) to nondiabetic and streptozotocin-induced diabetic male Wistar rats for 4 and 8 weeks. The effects of ALA were assessed starting from structural/morphological alterations through changes that characterize profibrotic processes, to regulation of collagen gene expression in kidney. Here, we demonstrated that ALA improved systemic glucose and urea level, reduced formation of renal advanced glycation end products (AGEs), and maintained renal structural integrity in diabetic rats. However, profibrotic events provoked in diabetes were not alleviated by ALA since collagen synthesis/deposition and expression of transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) remained elevated in ALA-treated diabetic rats, especially after 8 weeks of diabetes onset. Moreover, 8 weeks treatment of nondiabetic rats with ALA led to the development of profibrotic features reflected in increased collagen synthesis/deposition. Besides the TGF-β1 downstream signaling, the additional mechanism underlying the upregulation of collagen IV in nondiabetic rats treated with ALA involves decreased DNA methylation of its promoter that could arise from increased Tet1 expression. These findings emphasize the therapeutic caution in the use of ALA, especially in patients with renal diabetic complication.",
publisher = "Hindawi Limited",
journal = "Oxidative Medicine and Cellular Longevity",
title = "α-Lipoic Acid Increases Collagen Synthesis and Deposition in Nondiabetic and Diabetic Rat Kidneys",
volume = "2021",
doi = "10.1155/2021/6669352",
pages = "6669352"
}

Grdović, N., Rajić, J., Arambašić Jovanović, J., Dinić, S., Tolić, A., Đorđević, M., Đorđević, M., Trifunović, S., Vidaković, M., Uskoković, A.,& Mihailović, M.. (2021). α-Lipoic Acid Increases Collagen Synthesis and Deposition in Nondiabetic and Diabetic Rat Kidneys. in Oxidative Medicine and Cellular Longevity
Hindawi Limited., 2021, 6669352.
https://doi.org/10.1155/2021/6669352

Grdović N, Rajić J, Arambašić Jovanović J, Dinić S, Tolić A, Đorđević M, Đorđević M, Trifunović S, Vidaković M, Uskoković A, Mihailović M. α-Lipoic Acid Increases Collagen Synthesis and Deposition in Nondiabetic and Diabetic Rat Kidneys. in Oxidative Medicine and Cellular Longevity. 2021;2021:6669352.
doi:10.1155/2021/6669352 .

Grdović, Nevena, Rajić, Jovana, Arambašić Jovanović, Jelena, Dinić, Svetlana, Tolić, Anja, Đorđević, Miloš, Đorđević, Marija, Trifunović, Svetlana, Vidaković, Melita, Uskoković, Aleksandra, Mihailović, Mirjana, "α-Lipoic Acid Increases Collagen Synthesis and Deposition in Nondiabetic and Diabetic Rat Kidneys" in Oxidative Medicine and Cellular Longevity, 2021 (2021):6669352,
https://doi.org/10.1155/2021/6669352 . .

TY  - CONF
AU  - Jakubek, Patrycja
AU  - Rajić, Jovana
AU  - Baranowska, Monika
AU  - Vidaković, Melita
AU  - Namiesnik, Jacek
AU  - Bartoszek, Agnieszka
PY  - 2020
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6309
AB  - In our previous study, treatment of HT29 cell line with catechins induced dose-dependent changes in the expression of redox-related genes. Uniquely, only one gene (SRXN1, sulfiredoxin-1) was down-regulated upon treatment with 10 μM (-)-epigallocatechin (EGC). The aim of the current study was to investigate whether the observed down-regulation of SRXN1 expression was affected by epigenetic changes. HT29 cells were treated with catechins at different concentrations for 24 h and subsequently genomic DNA was isolated and bisulfite converted. DNA methylation profiles of selected regions within SRXN1 promoter were examined using Methylation-Specific PCR (MSP) and Methylation-Sensitive High Resolution Melting (MS-HRM). MSP analysis showed no differences in DNA methylation level between any of the treatments compared to control. However, the difference was observed when the bigger area of CpG island was analyzed by MS-HRM. Significant increase in DNA methylation level was observed after cell treatment with higher doses of EGC and (-)-epicatechin gallate (ECG). DNA demethylation requires oxidative modifications of methylated cytosine. Catechins, which are strong antioxidants, may lead to inhibition of DNA demethylation by changing cellular environment to more reduced state, especially in the case of higher doses. Thus, we report that catechins can act as methylation inducers and probably this function derives from their ability to influence cellular redox homeostasis.
PB  - Belgrade: NutRedox, COST Action CA16112
C3  - Nutraceuticals in balancing redox status in ageing and age-related diseases: Book of Abstracts. WGs Meeting of the NutRedOx COST Action CA16112; 2020 Mar 2-3; Belgrade, Serbia
T1  - The impact of catechins on DNA methylation level within promoter area of sulfiredoxin-1 gene in HT29 cell line
SP  - 25
EP  - 25
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6309
ER  - 

@conference{
editor = "Dinić, Svetlana, Šunderić, Miloš, Vučić, Vesna, Vidović, Bojana",
author = "Jakubek, Patrycja and Rajić, Jovana and Baranowska, Monika and Vidaković, Melita and Namiesnik, Jacek and Bartoszek, Agnieszka",
year = "2020",
abstract = "In our previous study, treatment of HT29 cell line with catechins induced dose-dependent changes in the expression of redox-related genes. Uniquely, only one gene (SRXN1, sulfiredoxin-1) was down-regulated upon treatment with 10 μM (-)-epigallocatechin (EGC). The aim of the current study was to investigate whether the observed down-regulation of SRXN1 expression was affected by epigenetic changes. HT29 cells were treated with catechins at different concentrations for 24 h and subsequently genomic DNA was isolated and bisulfite converted. DNA methylation profiles of selected regions within SRXN1 promoter were examined using Methylation-Specific PCR (MSP) and Methylation-Sensitive High Resolution Melting (MS-HRM). MSP analysis showed no differences in DNA methylation level between any of the treatments compared to control. However, the difference was observed when the bigger area of CpG island was analyzed by MS-HRM. Significant increase in DNA methylation level was observed after cell treatment with higher doses of EGC and (-)-epicatechin gallate (ECG). DNA demethylation requires oxidative modifications of methylated cytosine. Catechins, which are strong antioxidants, may lead to inhibition of DNA demethylation by changing cellular environment to more reduced state, especially in the case of higher doses. Thus, we report that catechins can act as methylation inducers and probably this function derives from their ability to influence cellular redox homeostasis.",
publisher = "Belgrade: NutRedox, COST Action CA16112",
journal = "Nutraceuticals in balancing redox status in ageing and age-related diseases: Book of Abstracts. WGs Meeting of the NutRedOx COST Action CA16112; 2020 Mar 2-3; Belgrade, Serbia",
title = "The impact of catechins on DNA methylation level within promoter area of sulfiredoxin-1 gene in HT29 cell line",
pages = "25-25",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6309"
}

Dinić, S., Šunderić, M., Vučić, V., Vidović, B., Jakubek, P., Rajić, J., Baranowska, M., Vidaković, M., Namiesnik, J.,& Bartoszek, A.. (2020). The impact of catechins on DNA methylation level within promoter area of sulfiredoxin-1 gene in HT29 cell line. in Nutraceuticals in balancing redox status in ageing and age-related diseases: Book of Abstracts. WGs Meeting of the NutRedOx COST Action CA16112; 2020 Mar 2-3; Belgrade, Serbia
Belgrade: NutRedox, COST Action CA16112., 25-25.
https://hdl.handle.net/21.15107/rcub_ibiss_6309

Dinić S, Šunderić M, Vučić V, Vidović B, Jakubek P, Rajić J, Baranowska M, Vidaković M, Namiesnik J, Bartoszek A. The impact of catechins on DNA methylation level within promoter area of sulfiredoxin-1 gene in HT29 cell line. in Nutraceuticals in balancing redox status in ageing and age-related diseases: Book of Abstracts. WGs Meeting of the NutRedOx COST Action CA16112; 2020 Mar 2-3; Belgrade, Serbia. 2020;:25-25.
https://hdl.handle.net/21.15107/rcub_ibiss_6309 .

Dinić, Svetlana, Šunderić, Miloš, Vučić, Vesna, Vidović, Bojana, Jakubek, Patrycja, Rajić, Jovana, Baranowska, Monika, Vidaković, Melita, Namiesnik, Jacek, Bartoszek, Agnieszka, "The impact of catechins on DNA methylation level within promoter area of sulfiredoxin-1 gene in HT29 cell line" in Nutraceuticals in balancing redox status in ageing and age-related diseases: Book of Abstracts. WGs Meeting of the NutRedOx COST Action CA16112; 2020 Mar 2-3; Belgrade, Serbia (2020):25-25,
https://hdl.handle.net/21.15107/rcub_ibiss_6309 .

TY  - JOUR
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Poznanović, Goran
AU  - Mihailović, Mirjana
AU  - Inic-Kanada, Aleksandra
AU  - Barisani-Asenbauer, Talin
AU  - Grdović, Nevena
AU  - Vidaković, Melita
PY  - 2020
UR  - http://www.ncbi.nlm.nih.gov/pubmed/32387379
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3687
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3695
AB  - Epithelial to mesenchymal transition (EMT) contributes to fibrosis associated pathologies including scarring of different ocular tissues. Recently targeting EMT is seen as an appropriate therapeutic approach for different fibrosis related eye diseases such as macular degeneration or glaucoma surgery related fibrosis. Nevertheless, for ocular surface diseases, target genes specific for particular cell type or condition are still undefined. This study aimed to expose the complex regulatory mechanisms that trigger EMT in human conjunctival epithelial (HCjE) cells. EMT was induced by prolonged treatment with two TGF-β isoforms, TGF-β1 and TGF-β2, and their combination. TGF-β1 showed the strongest potential for initiating EMT in HCjE cells, reflected on morphological changes, cell migration and the levels of mRNA expression of different epithelial (CDH1, OCLN, DSP) and mesenchymal (CDH2, FN1, VIM, SNAI1, ZEB2, TWIST1) marker genes. Co-treatment with the DNA demethylating agent 5-Azacytidine (5-AzaC) was capable of stopping the transition of HCjE cells towards a mesenchymal phenotype, based on morphological features, reduced cell mobility and mRNA and protein expression levels of epithelial and mesenchymal marker genes. An EMT qRT-PCR-based array revealed that EMT induced considerable alterations in gene expression, with downregulation of the majority of epithelial marker genes and upregulation of genes specific for the mesenchymal state. The major effect of 5-AzaC treatment was observed as a suppression of mesenchymal marker genes, suggesting the involvement of upstream negative regulator(s) whose promoter demethylation and subsequent expression will in turn promote EMT switch off. The expression level of miRNAs potentially important for EMT induction was determined using qRT-PCR-based array which pointed at members of miR-200 family as main regulators of EMT process in HCjE cells. 5-AzaC treatment induced increased expression of miR-200a, -200b, -200c and miR-141 towards the control level, indicating important role of DNA methylation in their regulation. The DNA methylation status of both miR-200 family clusters, analyzed with high-resolution melting (HRM) and bisulfite sequencing (Bis-Seq), revealed that TGF-β1-induced EMT was accompanied by increase in promoter CpG methylation of both miR-200 loci, which was reverted after 5-AzaC treatment. In conclusion, our results indicate that DNA demethylation of promoters of miR-200 loci is critically important for stopping and reverting the EMT in human conjunctival epithelial cells, suggesting the potential for the development of novel epigenetic-based therapeutic strategies for treating conjunctival conditions associated with EMT.
PB  - Elsevier BV
T2  - Experimental Eye Research
T1  - DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.
VL  - 197
DO  - 10.1016/j.exer.2020.108047
SP  - 108047
ER  - 

@article{
author = "Rajić, Jovana and Dinić, Svetlana and Uskoković, Aleksandra and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Poznanović, Goran and Mihailović, Mirjana and Inic-Kanada, Aleksandra and Barisani-Asenbauer, Talin and Grdović, Nevena and Vidaković, Melita",
year = "2020",
abstract = "Epithelial to mesenchymal transition (EMT) contributes to fibrosis associated pathologies including scarring of different ocular tissues. Recently targeting EMT is seen as an appropriate therapeutic approach for different fibrosis related eye diseases such as macular degeneration or glaucoma surgery related fibrosis. Nevertheless, for ocular surface diseases, target genes specific for particular cell type or condition are still undefined. This study aimed to expose the complex regulatory mechanisms that trigger EMT in human conjunctival epithelial (HCjE) cells. EMT was induced by prolonged treatment with two TGF-β isoforms, TGF-β1 and TGF-β2, and their combination. TGF-β1 showed the strongest potential for initiating EMT in HCjE cells, reflected on morphological changes, cell migration and the levels of mRNA expression of different epithelial (CDH1, OCLN, DSP) and mesenchymal (CDH2, FN1, VIM, SNAI1, ZEB2, TWIST1) marker genes. Co-treatment with the DNA demethylating agent 5-Azacytidine (5-AzaC) was capable of stopping the transition of HCjE cells towards a mesenchymal phenotype, based on morphological features, reduced cell mobility and mRNA and protein expression levels of epithelial and mesenchymal marker genes. An EMT qRT-PCR-based array revealed that EMT induced considerable alterations in gene expression, with downregulation of the majority of epithelial marker genes and upregulation of genes specific for the mesenchymal state. The major effect of 5-AzaC treatment was observed as a suppression of mesenchymal marker genes, suggesting the involvement of upstream negative regulator(s) whose promoter demethylation and subsequent expression will in turn promote EMT switch off. The expression level of miRNAs potentially important for EMT induction was determined using qRT-PCR-based array which pointed at members of miR-200 family as main regulators of EMT process in HCjE cells. 5-AzaC treatment induced increased expression of miR-200a, -200b, -200c and miR-141 towards the control level, indicating important role of DNA methylation in their regulation. The DNA methylation status of both miR-200 family clusters, analyzed with high-resolution melting (HRM) and bisulfite sequencing (Bis-Seq), revealed that TGF-β1-induced EMT was accompanied by increase in promoter CpG methylation of both miR-200 loci, which was reverted after 5-AzaC treatment. In conclusion, our results indicate that DNA demethylation of promoters of miR-200 loci is critically important for stopping and reverting the EMT in human conjunctival epithelial cells, suggesting the potential for the development of novel epigenetic-based therapeutic strategies for treating conjunctival conditions associated with EMT.",
publisher = "Elsevier BV",
journal = "Experimental Eye Research",
title = "DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.",
volume = "197",
doi = "10.1016/j.exer.2020.108047",
pages = "108047"
}

Rajić, J., Dinić, S., Uskoković, A., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Đorđević, M., Poznanović, G., Mihailović, M., Inic-Kanada, A., Barisani-Asenbauer, T., Grdović, N.,& Vidaković, M.. (2020). DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.. in Experimental Eye Research
Elsevier BV., 197, 108047.
https://doi.org/10.1016/j.exer.2020.108047

Rajić J, Dinić S, Uskoković A, Arambašić Jovanović J, Tolić A, Đorđević M, Đorđević M, Poznanović G, Mihailović M, Inic-Kanada A, Barisani-Asenbauer T, Grdović N, Vidaković M. DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.. in Experimental Eye Research. 2020;197:108047.
doi:10.1016/j.exer.2020.108047 .

Rajić, Jovana, Dinić, Svetlana, Uskoković, Aleksandra, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Poznanović, Goran, Mihailović, Mirjana, Inic-Kanada, Aleksandra, Barisani-Asenbauer, Talin, Grdović, Nevena, Vidaković, Melita, "DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells." in Experimental Eye Research, 197 (2020):108047,
https://doi.org/10.1016/j.exer.2020.108047 . .

TY  - JOUR
AU  - Rajić, Jovana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Poznanović, Goran
AU  - Mihailović, Mirjana
AU  - Inic-Kanada, Aleksandra
AU  - Barisani-Asenbauer, Talin
AU  - Grdović, Nevena
AU  - Vidaković, Melita
PY  - 2020
UR  - http://www.ncbi.nlm.nih.gov/pubmed/32387379
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3687
UR  - https://radar.ibiss.bg.ac.rs/handle/handle/123456789/3695
AB  - Epithelial to mesenchymal transition (EMT) contributes to fibrosis associated pathologies including scarring of different ocular tissues. Recently targeting EMT is seen as an appropriate therapeutic approach for different fibrosis related eye diseases such as macular degeneration or glaucoma surgery related fibrosis. Nevertheless, for ocular surface diseases, target genes specific for particular cell type or condition are still undefined. This study aimed to expose the complex regulatory mechanisms that trigger EMT in human conjunctival epithelial (HCjE) cells. EMT was induced by prolonged treatment with two TGF-β isoforms, TGF-β1 and TGF-β2, and their combination. TGF-β1 showed the strongest potential for initiating EMT in HCjE cells, reflected on morphological changes, cell migration and the levels of mRNA expression of different epithelial (CDH1, OCLN, DSP) and mesenchymal (CDH2, FN1, VIM, SNAI1, ZEB2, TWIST1) marker genes. Co-treatment with the DNA demethylating agent 5-Azacytidine (5-AzaC) was capable of stopping the transition of HCjE cells towards a mesenchymal phenotype, based on morphological features, reduced cell mobility and mRNA and protein expression levels of epithelial and mesenchymal marker genes. An EMT qRT-PCR-based array revealed that EMT induced considerable alterations in gene expression, with downregulation of the majority of epithelial marker genes and upregulation of genes specific for the mesenchymal state. The major effect of 5-AzaC treatment was observed as a suppression of mesenchymal marker genes, suggesting the involvement of upstream negative regulator(s) whose promoter demethylation and subsequent expression will in turn promote EMT switch off. The expression level of miRNAs potentially important for EMT induction was determined using qRT-PCR-based array which pointed at members of miR-200 family as main regulators of EMT process in HCjE cells. 5-AzaC treatment induced increased expression of miR-200a, -200b, -200c and miR-141 towards the control level, indicating important role of DNA methylation in their regulation. The DNA methylation status of both miR-200 family clusters, analyzed with high-resolution melting (HRM) and bisulfite sequencing (Bis-Seq), revealed that TGF-β1-induced EMT was accompanied by increase in promoter CpG methylation of both miR-200 loci, which was reverted after 5-AzaC treatment. In conclusion, our results indicate that DNA demethylation of promoters of miR-200 loci is critically important for stopping and reverting the EMT in human conjunctival epithelial cells, suggesting the potential for the development of novel epigenetic-based therapeutic strategies for treating conjunctival conditions associated with EMT.
PB  - Elsevier BV
T2  - Experimental Eye Research
T1  - DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.
VL  - 197
DO  - 10.1016/j.exer.2020.108047
SP  - 108047
ER  - 

@article{
author = "Rajić, Jovana and Dinić, Svetlana and Uskoković, Aleksandra and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Poznanović, Goran and Mihailović, Mirjana and Inic-Kanada, Aleksandra and Barisani-Asenbauer, Talin and Grdović, Nevena and Vidaković, Melita",
year = "2020",
abstract = "Epithelial to mesenchymal transition (EMT) contributes to fibrosis associated pathologies including scarring of different ocular tissues. Recently targeting EMT is seen as an appropriate therapeutic approach for different fibrosis related eye diseases such as macular degeneration or glaucoma surgery related fibrosis. Nevertheless, for ocular surface diseases, target genes specific for particular cell type or condition are still undefined. This study aimed to expose the complex regulatory mechanisms that trigger EMT in human conjunctival epithelial (HCjE) cells. EMT was induced by prolonged treatment with two TGF-β isoforms, TGF-β1 and TGF-β2, and their combination. TGF-β1 showed the strongest potential for initiating EMT in HCjE cells, reflected on morphological changes, cell migration and the levels of mRNA expression of different epithelial (CDH1, OCLN, DSP) and mesenchymal (CDH2, FN1, VIM, SNAI1, ZEB2, TWIST1) marker genes. Co-treatment with the DNA demethylating agent 5-Azacytidine (5-AzaC) was capable of stopping the transition of HCjE cells towards a mesenchymal phenotype, based on morphological features, reduced cell mobility and mRNA and protein expression levels of epithelial and mesenchymal marker genes. An EMT qRT-PCR-based array revealed that EMT induced considerable alterations in gene expression, with downregulation of the majority of epithelial marker genes and upregulation of genes specific for the mesenchymal state. The major effect of 5-AzaC treatment was observed as a suppression of mesenchymal marker genes, suggesting the involvement of upstream negative regulator(s) whose promoter demethylation and subsequent expression will in turn promote EMT switch off. The expression level of miRNAs potentially important for EMT induction was determined using qRT-PCR-based array which pointed at members of miR-200 family as main regulators of EMT process in HCjE cells. 5-AzaC treatment induced increased expression of miR-200a, -200b, -200c and miR-141 towards the control level, indicating important role of DNA methylation in their regulation. The DNA methylation status of both miR-200 family clusters, analyzed with high-resolution melting (HRM) and bisulfite sequencing (Bis-Seq), revealed that TGF-β1-induced EMT was accompanied by increase in promoter CpG methylation of both miR-200 loci, which was reverted after 5-AzaC treatment. In conclusion, our results indicate that DNA demethylation of promoters of miR-200 loci is critically important for stopping and reverting the EMT in human conjunctival epithelial cells, suggesting the potential for the development of novel epigenetic-based therapeutic strategies for treating conjunctival conditions associated with EMT.",
publisher = "Elsevier BV",
journal = "Experimental Eye Research",
title = "DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.",
volume = "197",
doi = "10.1016/j.exer.2020.108047",
pages = "108047"
}

Rajić, J., Dinić, S., Uskoković, A., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Đorđević, M., Poznanović, G., Mihailović, M., Inic-Kanada, A., Barisani-Asenbauer, T., Grdović, N.,& Vidaković, M.. (2020). DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.. in Experimental Eye Research
Elsevier BV., 197, 108047.
https://doi.org/10.1016/j.exer.2020.108047

Rajić J, Dinić S, Uskoković A, Arambašić Jovanović J, Tolić A, Đorđević M, Đorđević M, Poznanović G, Mihailović M, Inic-Kanada A, Barisani-Asenbauer T, Grdović N, Vidaković M. DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells.. in Experimental Eye Research. 2020;197:108047.
doi:10.1016/j.exer.2020.108047 .

Rajić, Jovana, Dinić, Svetlana, Uskoković, Aleksandra, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Poznanović, Goran, Mihailović, Mirjana, Inic-Kanada, Aleksandra, Barisani-Asenbauer, Talin, Grdović, Nevena, Vidaković, Melita, "DNA methylation of miR-200 clusters promotes epithelial to mesenchymal transition in human conjunctival epithelial cells." in Experimental Eye Research, 197 (2020):108047,
https://doi.org/10.1016/j.exer.2020.108047 . .

TY  - JOUR
AU  - Đorđević, Miloš
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Uskoković, Aleksandra
AU  - Rajić, Jovana
AU  - Đorđević, Marija
AU  - Tolić, Anja
AU  - Mišić, Danijela
AU  - Šiler, Branislav
AU  - Poznanović, Goran
AU  - Vidaković, Melita
AU  - Dinić, Svetlana
PY  - 2020
UR  - http://www.serbiosoc.org.rs/arch/index.php/abs/article/view/5029
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3651
AB  - Oxidative stress is one of the major mechanisms that underlies the damage of pancreatic b-cells and defects in insulin secretion in diabetes. As herbal preparations can alleviate oxidative stress through their redox-active secondary metabolites, in this study we investigated the cytoprotective effects of Centaurium erythraea extract (CEe) against H2O2- and SNP-induced oxidative/nitrosative stress in Rin-5F b-cells. The antioxidant activity of CEe and its effect on cell survival and insulin expression/secretion were evaluated. The CEe increased cell viability and ameliorated the disturbance of redox homeostasis in H2O2- and SNP-treated cells by decreasing DNA damage, lipid peroxidation and protein S-glutathionylation. The CEe restored GSH homeostasis in H2O2-treated b-cells and attenuated the SNP-induced disturbance of the GSH/GSSG ratio. The H2O2- and SNP-induced disruption of CAT, GPx, GR, MnSOD and CuZnSOD activities was adjusted by the CEe towards control values, as well as mRNA and protein levels of GPx, MnSOD and CAT. The CEe increased insulin expression/secretion particularly in H2O2-treated b-cells, which was in accordance with the more pronounced antioxidant effect of the CEe observed in H2O2-treated b-cells as compared to SNP-treated cells. These findings support the beneficial effect of the CEe in preventing or slowing down b-cell damage and dysfunction caused by oxidative/nitrosative stress during diabetes development.
T2  - Archives of Biological Sciences
T1  - Centaurium erythraea extract reduces redox imbalance and improves insulin expression and secretion in pancreatic β-cells exposed to oxidative and nitrosative stress
IS  - 1
VL  - 72
DO  - 10.2298/abs200127005d
SP  - 117
EP  - 128
ER  - 

@article{
author = "Đorđević, Miloš and Grdović, Nevena and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Uskoković, Aleksandra and Rajić, Jovana and Đorđević, Marija and Tolić, Anja and Mišić, Danijela and Šiler, Branislav and Poznanović, Goran and Vidaković, Melita and Dinić, Svetlana",
year = "2020",
abstract = "Oxidative stress is one of the major mechanisms that underlies the damage of pancreatic b-cells and defects in insulin secretion in diabetes. As herbal preparations can alleviate oxidative stress through their redox-active secondary metabolites, in this study we investigated the cytoprotective effects of Centaurium erythraea extract (CEe) against H2O2- and SNP-induced oxidative/nitrosative stress in Rin-5F b-cells. The antioxidant activity of CEe and its effect on cell survival and insulin expression/secretion were evaluated. The CEe increased cell viability and ameliorated the disturbance of redox homeostasis in H2O2- and SNP-treated cells by decreasing DNA damage, lipid peroxidation and protein S-glutathionylation. The CEe restored GSH homeostasis in H2O2-treated b-cells and attenuated the SNP-induced disturbance of the GSH/GSSG ratio. The H2O2- and SNP-induced disruption of CAT, GPx, GR, MnSOD and CuZnSOD activities was adjusted by the CEe towards control values, as well as mRNA and protein levels of GPx, MnSOD and CAT. The CEe increased insulin expression/secretion particularly in H2O2-treated b-cells, which was in accordance with the more pronounced antioxidant effect of the CEe observed in H2O2-treated b-cells as compared to SNP-treated cells. These findings support the beneficial effect of the CEe in preventing or slowing down b-cell damage and dysfunction caused by oxidative/nitrosative stress during diabetes development.",
journal = "Archives of Biological Sciences",
title = "Centaurium erythraea extract reduces redox imbalance and improves insulin expression and secretion in pancreatic β-cells exposed to oxidative and nitrosative stress",
number = "1",
volume = "72",
doi = "10.2298/abs200127005d",
pages = "117-128"
}

Đorđević, M., Grdović, N., Mihailović, M., Arambašić Jovanović, J., Uskoković, A., Rajić, J., Đorđević, M., Tolić, A., Mišić, D., Šiler, B., Poznanović, G., Vidaković, M.,& Dinić, S.. (2020). Centaurium erythraea extract reduces redox imbalance and improves insulin expression and secretion in pancreatic β-cells exposed to oxidative and nitrosative stress. in Archives of Biological Sciences, 72(1), 117-128.
https://doi.org/10.2298/abs200127005d

Đorđević M, Grdović N, Mihailović M, Arambašić Jovanović J, Uskoković A, Rajić J, Đorđević M, Tolić A, Mišić D, Šiler B, Poznanović G, Vidaković M, Dinić S. Centaurium erythraea extract reduces redox imbalance and improves insulin expression and secretion in pancreatic β-cells exposed to oxidative and nitrosative stress. in Archives of Biological Sciences. 2020;72(1):117-128.
doi:10.2298/abs200127005d .

Đorđević, Miloš, Grdović, Nevena, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Uskoković, Aleksandra, Rajić, Jovana, Đorđević, Marija, Tolić, Anja, Mišić, Danijela, Šiler, Branislav, Poznanović, Goran, Vidaković, Melita, Dinić, Svetlana, "Centaurium erythraea extract reduces redox imbalance and improves insulin expression and secretion in pancreatic β-cells exposed to oxidative and nitrosative stress" in Archives of Biological Sciences, 72, no. 1 (2020):117-128,
https://doi.org/10.2298/abs200127005d . .

TY  - CONF
AU  - Đorđević, Miloš
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Uskoković, Aleksandra
AU  - Rajić, Jovana
AU  - Đorđević, Marija
AU  - Tolić, Anja
AU  - Mišić, Danijela
AU  - Šiler, Branislav
AU  - Poznanović, Goran
AU  - Vidaković, Melita
AU  - Dinić, Svetlana
PY  - 2020
UR  - https://blog.u-bourgogne.fr/cost-nutredox/wp-content/uploads/sites/81/2020/03/NutRedOX-Belgrade-2020-Abstract-book.pdf
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4107
AB  - Centaurium erythraea (CE) is traditionally used for diabetes treatment due to its anti-diabetic properties. Previously we have reported that the major constituents of CE methanol extract (CEE) are secoiridoids and polyphenols. Here we analyzed the protective effect of CEE on pancreatic β-cells in streptozotocin (STZ)-induced diabetic rats. CEE (100 mg/kg) was administered daily and orally to control or diabetic rats for two weeks before diabetes induction, during five days of STZ treatment (40 mg/kg/day), and for four weeks after last STZ injection (pre-treated group), or for four weeks after diabetes induction (post-treated group). Histological and immunohistochemical examination of the pancreas revealed disturbed morphology of pancreatic islets, a decrease in their number and size which was accompanied by the reduction of insulin-positive β-cells in diabetic rats when compared to control or control/CEE-treated rats. Islet morphology and mass, as well as the number of insulin-positive β-cells, were improved in CEE-treated diabetic rats, particularly in a pre-treated group. In pre- and post-treated groups we observed the increase of GLUT-2 transporter and p-Akt kinase, that were absent in diabetic pancreas pointing to impaired glucose-stimulated insulin secretion in remnant β-cells. CEE-mediated increase of β-cell mass, GLUT-2 and p-Akt levels in diabetic rats contributed to the elevation of serum insulin level and reduction of glucose level which was more prominent in pre- than in a post-treated group. The results of this study suggest that improved insulin production and glycemic control in CEE-treated diabetic rats may result from the structural/functional preservation of pancreatic islets.
PB  - NutRedox, COST Action CA16112
C3  - WGs Meeting of the NutRedOx COST Action CA16112 Belgrade, March 2-3, 2020
T1  - Beneficial effects of Centaurium erythraea extract on glycemic control and insulin  level in diabetic rats
SP  - 9
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4107
ER  - 

@conference{
author = "Đorđević, Miloš and Grdović, Nevena and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Uskoković, Aleksandra and Rajić, Jovana and Đorđević, Marija and Tolić, Anja and Mišić, Danijela and Šiler, Branislav and Poznanović, Goran and Vidaković, Melita and Dinić, Svetlana",
year = "2020",
abstract = "Centaurium erythraea (CE) is traditionally used for diabetes treatment due to its anti-diabetic properties. Previously we have reported that the major constituents of CE methanol extract (CEE) are secoiridoids and polyphenols. Here we analyzed the protective effect of CEE on pancreatic β-cells in streptozotocin (STZ)-induced diabetic rats. CEE (100 mg/kg) was administered daily and orally to control or diabetic rats for two weeks before diabetes induction, during five days of STZ treatment (40 mg/kg/day), and for four weeks after last STZ injection (pre-treated group), or for four weeks after diabetes induction (post-treated group). Histological and immunohistochemical examination of the pancreas revealed disturbed morphology of pancreatic islets, a decrease in their number and size which was accompanied by the reduction of insulin-positive β-cells in diabetic rats when compared to control or control/CEE-treated rats. Islet morphology and mass, as well as the number of insulin-positive β-cells, were improved in CEE-treated diabetic rats, particularly in a pre-treated group. In pre- and post-treated groups we observed the increase of GLUT-2 transporter and p-Akt kinase, that were absent in diabetic pancreas pointing to impaired glucose-stimulated insulin secretion in remnant β-cells. CEE-mediated increase of β-cell mass, GLUT-2 and p-Akt levels in diabetic rats contributed to the elevation of serum insulin level and reduction of glucose level which was more prominent in pre- than in a post-treated group. The results of this study suggest that improved insulin production and glycemic control in CEE-treated diabetic rats may result from the structural/functional preservation of pancreatic islets.",
publisher = "NutRedox, COST Action CA16112",
journal = "WGs Meeting of the NutRedOx COST Action CA16112 Belgrade, March 2-3, 2020",
title = "Beneficial effects of Centaurium erythraea extract on glycemic control and insulin  level in diabetic rats",
pages = "9",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4107"
}

Đorđević, M., Grdović, N., Mihailović, M., Arambašić Jovanović, J., Uskoković, A., Rajić, J., Đorđević, M., Tolić, A., Mišić, D., Šiler, B., Poznanović, G., Vidaković, M.,& Dinić, S.. (2020). Beneficial effects of Centaurium erythraea extract on glycemic control and insulin  level in diabetic rats. in WGs Meeting of the NutRedOx COST Action CA16112 Belgrade, March 2-3, 2020
NutRedox, COST Action CA16112., 9.
https://hdl.handle.net/21.15107/rcub_ibiss_4107

Đorđević M, Grdović N, Mihailović M, Arambašić Jovanović J, Uskoković A, Rajić J, Đorđević M, Tolić A, Mišić D, Šiler B, Poznanović G, Vidaković M, Dinić S. Beneficial effects of Centaurium erythraea extract on glycemic control and insulin  level in diabetic rats. in WGs Meeting of the NutRedOx COST Action CA16112 Belgrade, March 2-3, 2020. 2020;:9.
https://hdl.handle.net/21.15107/rcub_ibiss_4107 .

Đorđević, Miloš, Grdović, Nevena, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Uskoković, Aleksandra, Rajić, Jovana, Đorđević, Marija, Tolić, Anja, Mišić, Danijela, Šiler, Branislav, Poznanović, Goran, Vidaković, Melita, Dinić, Svetlana, "Beneficial effects of Centaurium erythraea extract on glycemic control and insulin  level in diabetic rats" in WGs Meeting of the NutRedOx COST Action CA16112 Belgrade, March 2-3, 2020 (2020):9,
https://hdl.handle.net/21.15107/rcub_ibiss_4107 .

TY  - CONF
AU  - Tolić, Anja
AU  - Rajić, Jovana
AU  - Đorđević, Miloš
AU  - Đorđević, Marija
AU  - Uskoković, Aleksandra
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Dinić, Svetlana
AU  - Nestorović, Nataša
AU  - Jurkowski, Tomasz
AU  - Vidaković, Melita
PY  - 2020
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/4783
AB  - Since our previous work indicated interplay of Ten-eleven translocation (TET) and Poly (ADP-ribose) polymerase (PARP) proteins our aim was to further study the effects of their interaction. 
The ability of recombinant PARP-1 to poly ADP-ribosylate (PARylate) catalytic domains of TET proteins was examined in vitro. It was observed that all TETs (TET1, TET2, TET3) readily undergo PARylation. To our knowledge, this is the first report of PARP-1 PARylation of TET2 and TET3 while TET1 PARylation has been previously documented. PARylation of TET proteins was evidenced by western blot signal stretching from the position of unmodified TET. This type of signal is characteristic for PARylation since proteins are modified to varying degrees by covalently attached negatively charged PAR polymers. This slows their movement during electrophoresis and individual molecules are expected to stop at different positions resulting in signal stretching upwards.  PARylation can introduce electrostatic and topological changes in modified proteins, resulting in altered enzymatic activity. Therefore we evaluated TET1 activity in vitro using an ELISA type in-house assay, which showed that PARP-1-dependent PARylation lowers the ability of TET1 catalytic domain to oxidize 5mC to 5hmC. 
To corroborate these results in cellulo, we examined changes in DNA methylation in mouse embryonic fibroblasts (NIH3T3) compared to a PARP-1 knock out mouse embryonic fibroblast cell line (P ARP-/-). Lower methylation was observed in PARP-/- cells by immunocytochemical staining. Next, we analyzed global DNA methylation by ELISA assay and we again detected lower methylation levels in PARP-/- and also in NIH3T3 cells treated by a PARP inhibitor niraparib. Finally, DNA hydroxymethylation was assessed by immunocytochemistry and stronger signal was observed in PARP-/- cells and NIH3T3 cells treated by niraparib, compared to control NIH3T3 cells. 
In summary, inhibition of PARylation or absence of PARP-1 lead to decreased methylation and increased hydroxymethylation of DNA in cellulo. Together, our results point to the inhibitory influence that PARP-1 and PARylation exert on TET1 activity. Further studies are needed to evaluate the effects of PARylation of other TET proteins as well as to examine potential influence of other PARPs.
PB  - Cold Spring Harbor Laboratory
C3  - 2020 virtual meeting on EPIGENETICS & CHROMATIN; 2020 Sep 15-18; Virtual meeting
T1  - PARP-1 and PARylation inhibit TET1 demethylation activity
SP  - 286
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4783
ER  - 

@conference{
author = "Tolić, Anja and Rajić, Jovana and Đorđević, Miloš and Đorđević, Marija and Uskoković, Aleksandra and Grdović, Nevena and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Dinić, Svetlana and Nestorović, Nataša and Jurkowski, Tomasz and Vidaković, Melita",
year = "2020",
abstract = "Since our previous work indicated interplay of Ten-eleven translocation (TET) and Poly (ADP-ribose) polymerase (PARP) proteins our aim was to further study the effects of their interaction. 
The ability of recombinant PARP-1 to poly ADP-ribosylate (PARylate) catalytic domains of TET proteins was examined in vitro. It was observed that all TETs (TET1, TET2, TET3) readily undergo PARylation. To our knowledge, this is the first report of PARP-1 PARylation of TET2 and TET3 while TET1 PARylation has been previously documented. PARylation of TET proteins was evidenced by western blot signal stretching from the position of unmodified TET. This type of signal is characteristic for PARylation since proteins are modified to varying degrees by covalently attached negatively charged PAR polymers. This slows their movement during electrophoresis and individual molecules are expected to stop at different positions resulting in signal stretching upwards.  PARylation can introduce electrostatic and topological changes in modified proteins, resulting in altered enzymatic activity. Therefore we evaluated TET1 activity in vitro using an ELISA type in-house assay, which showed that PARP-1-dependent PARylation lowers the ability of TET1 catalytic domain to oxidize 5mC to 5hmC. 
To corroborate these results in cellulo, we examined changes in DNA methylation in mouse embryonic fibroblasts (NIH3T3) compared to a PARP-1 knock out mouse embryonic fibroblast cell line (P ARP-/-). Lower methylation was observed in PARP-/- cells by immunocytochemical staining. Next, we analyzed global DNA methylation by ELISA assay and we again detected lower methylation levels in PARP-/- and also in NIH3T3 cells treated by a PARP inhibitor niraparib. Finally, DNA hydroxymethylation was assessed by immunocytochemistry and stronger signal was observed in PARP-/- cells and NIH3T3 cells treated by niraparib, compared to control NIH3T3 cells. 
In summary, inhibition of PARylation or absence of PARP-1 lead to decreased methylation and increased hydroxymethylation of DNA in cellulo. Together, our results point to the inhibitory influence that PARP-1 and PARylation exert on TET1 activity. Further studies are needed to evaluate the effects of PARylation of other TET proteins as well as to examine potential influence of other PARPs.",
publisher = "Cold Spring Harbor Laboratory",
journal = "2020 virtual meeting on EPIGENETICS & CHROMATIN; 2020 Sep 15-18; Virtual meeting",
title = "PARP-1 and PARylation inhibit TET1 demethylation activity",
pages = "286",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4783"
}

Tolić, A., Rajić, J., Đorđević, M., Đorđević, M., Uskoković, A., Grdović, N., Mihailović, M., Arambašić Jovanović, J., Dinić, S., Nestorović, N., Jurkowski, T.,& Vidaković, M.. (2020). PARP-1 and PARylation inhibit TET1 demethylation activity. in 2020 virtual meeting on EPIGENETICS & CHROMATIN; 2020 Sep 15-18; Virtual meeting
Cold Spring Harbor Laboratory., 286.
https://hdl.handle.net/21.15107/rcub_ibiss_4783

Tolić A, Rajić J, Đorđević M, Đorđević M, Uskoković A, Grdović N, Mihailović M, Arambašić Jovanović J, Dinić S, Nestorović N, Jurkowski T, Vidaković M. PARP-1 and PARylation inhibit TET1 demethylation activity. in 2020 virtual meeting on EPIGENETICS & CHROMATIN; 2020 Sep 15-18; Virtual meeting. 2020;:286.
https://hdl.handle.net/21.15107/rcub_ibiss_4783 .

Tolić, Anja, Rajić, Jovana, Đorđević, Miloš, Đorđević, Marija, Uskoković, Aleksandra, Grdović, Nevena, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Dinić, Svetlana, Nestorović, Nataša, Jurkowski, Tomasz, Vidaković, Melita, "PARP-1 and PARylation inhibit TET1 demethylation activity" in 2020 virtual meeting on EPIGENETICS & CHROMATIN; 2020 Sep 15-18; Virtual meeting (2020):286,
https://hdl.handle.net/21.15107/rcub_ibiss_4783 .

TY  - THES
AU  - Rajić, Jovana
PY  - 2020
UR  - https://radar.ibiss.bg.ac.rs/123456789/3884
AB  - Proces epitelno-mezenhimske tranzicije (EMT) može predstavljati značajan faktor koji doprinosi nastanku fibroznih promena konjuktive u različitim bolestima oka, ali je njegova uloga nedovoljno ispitana. Cilj ove doktorske disertacije podrazumevao je ispitivanje sposobnosti ćelijske linije humanih epitelnih ćelija konjuktive (HCjE) da uđu u proces EMT nakon infekcije uzročnikom trahoma, bakterijom Chlamydia trachomatis, i identifikaciju ključnih faktora povezanih sa pokretanjem i progresijom tranzicije na modelu procesa EMT indukovanog TGF-β proteinima. Posebna pažnja bila je usmerena na rasvetljavanje uloge metilacije DNK u regulaciji ekspresije gena uključenih u tranziciju HCjE ćelija. Praćenjem aktivacije signalnih puteva povezanih sa pokretanjem procesa EMT i ekspresije markera epitelnih/mezenhimskih ćelija pokazano je da HCjE ćelije poseduju sposobnost tranzicije nakon infekcije bakterijom C. trachomatis, praćene promenama u metilacionim profilima gena markera epitelnog/mezenhimskog fenotipa. Praćenjem morfologije, sposobnosti migracije i nivoa ekspresije gena markera uočeno je da dugotrajan tretman TGF-β1 izoformom ima najveći potencijal za pokretanje procesa EMT u HCjE ćelijama, dok su tretmani DNK demetilujućim agensom pokazali da metilacija DNK predstavlja važan mehanizam koji leži u osnovi ovog procesa. Promene profila metilacije DNK, detektovane analizom kriva topljenja i bisulfitnim sekvenciranjem, istakle su ključnu ulogu članova familije miR-200 u pokretanju i reverziji procesa EMT u HCjE ćelijama tretiranim TGF-β1 proteinima. Rezultati dobijeni u ovoj doktorskoj disertaciji ukazuju na mogućnost razvijanja novih selektivnih terapeutskih strategija u lečenju fibroznih promena konjuktive, zasnovanim na reverzibilnosti procesa EMT i epigenetičkih mehanizama.
AB  - The process of epithelial-mesenchymal transition (EMT) could be an important factor in development of fibrosis-related conjunctival eye diseases, but its precise role in these conditions has not been defined yet. The aim of this doctoral dissertation was to examine the ability of human epithelial conjunctival cell line (HCjE) to enter EMT process after infection with trachoma causative agent, bacteria Chlamydia trachomatis, i to identify key factors associated with the initiation i progression of transition in TGF-β-induced EMT model. Special focus was directed to elucidating the role of DNA methylation in the regulation of expression of genes involved in the transition of HCjE cells. Activation of signaling pathways associated with the initiation of the EMT process i the expression level of epithelial/mesenchymal markers revealed that HCjE cells possess the ability to enter the transition after infection with bacteria C. trachomatis, which was associated with changes in the methylation profiles of epithelial/mesenchymal gene markers. Cell morphology, migration ability i expression level of marker genes indicated that long-term treatment with TGF-β1 isoform has the greatest potential to initiate EMT process in HCjE cells, while treatments with DNA demethylating agent suggested that DNA methylation is an important mechanism which underlies this process. Changes in DNA methylation profile, detected by melting curve analysis i bisulfite sequencing, highlighted a key role of miR-200 family members in initiating i reversing TGF-β1-induced EMT process in HCjE cells. The results obtained in this doctoral dissertation indicate the possibility of developing new selective therapeutic strategies in the treatment of fibrosis-related conjunctival diseases, based on the reversibility of both EMT process i epigenetic mechanisms.
PB  - Belgrade: Faculty of Biology, University of Belgrade
T2  - Faculty of Biology, University of Belgrade
T1  - Uloga metilacije DNK u procesu epitelno-mezenhimske tranzicije  humanih epitelnih ćelija konjuktive
T1  - The role of DNA methylation in the process of epithelial-mesenchymal transition of human conjunctival epithelial cells
SP  - 1
EP  - 133
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_3884
ER  - 

@phdthesis{
author = "Rajić, Jovana",
year = "2020",
abstract = "Proces epitelno-mezenhimske tranzicije (EMT) može predstavljati značajan faktor koji doprinosi nastanku fibroznih promena konjuktive u različitim bolestima oka, ali je njegova uloga nedovoljno ispitana. Cilj ove doktorske disertacije podrazumevao je ispitivanje sposobnosti ćelijske linije humanih epitelnih ćelija konjuktive (HCjE) da uđu u proces EMT nakon infekcije uzročnikom trahoma, bakterijom Chlamydia trachomatis, i identifikaciju ključnih faktora povezanih sa pokretanjem i progresijom tranzicije na modelu procesa EMT indukovanog TGF-β proteinima. Posebna pažnja bila je usmerena na rasvetljavanje uloge metilacije DNK u regulaciji ekspresije gena uključenih u tranziciju HCjE ćelija. Praćenjem aktivacije signalnih puteva povezanih sa pokretanjem procesa EMT i ekspresije markera epitelnih/mezenhimskih ćelija pokazano je da HCjE ćelije poseduju sposobnost tranzicije nakon infekcije bakterijom C. trachomatis, praćene promenama u metilacionim profilima gena markera epitelnog/mezenhimskog fenotipa. Praćenjem morfologije, sposobnosti migracije i nivoa ekspresije gena markera uočeno je da dugotrajan tretman TGF-β1 izoformom ima najveći potencijal za pokretanje procesa EMT u HCjE ćelijama, dok su tretmani DNK demetilujućim agensom pokazali da metilacija DNK predstavlja važan mehanizam koji leži u osnovi ovog procesa. Promene profila metilacije DNK, detektovane analizom kriva topljenja i bisulfitnim sekvenciranjem, istakle su ključnu ulogu članova familije miR-200 u pokretanju i reverziji procesa EMT u HCjE ćelijama tretiranim TGF-β1 proteinima. Rezultati dobijeni u ovoj doktorskoj disertaciji ukazuju na mogućnost razvijanja novih selektivnih terapeutskih strategija u lečenju fibroznih promena konjuktive, zasnovanim na reverzibilnosti procesa EMT i epigenetičkih mehanizama., The process of epithelial-mesenchymal transition (EMT) could be an important factor in development of fibrosis-related conjunctival eye diseases, but its precise role in these conditions has not been defined yet. The aim of this doctoral dissertation was to examine the ability of human epithelial conjunctival cell line (HCjE) to enter EMT process after infection with trachoma causative agent, bacteria Chlamydia trachomatis, i to identify key factors associated with the initiation i progression of transition in TGF-β-induced EMT model. Special focus was directed to elucidating the role of DNA methylation in the regulation of expression of genes involved in the transition of HCjE cells. Activation of signaling pathways associated with the initiation of the EMT process i the expression level of epithelial/mesenchymal markers revealed that HCjE cells possess the ability to enter the transition after infection with bacteria C. trachomatis, which was associated with changes in the methylation profiles of epithelial/mesenchymal gene markers. Cell morphology, migration ability i expression level of marker genes indicated that long-term treatment with TGF-β1 isoform has the greatest potential to initiate EMT process in HCjE cells, while treatments with DNA demethylating agent suggested that DNA methylation is an important mechanism which underlies this process. Changes in DNA methylation profile, detected by melting curve analysis i bisulfite sequencing, highlighted a key role of miR-200 family members in initiating i reversing TGF-β1-induced EMT process in HCjE cells. The results obtained in this doctoral dissertation indicate the possibility of developing new selective therapeutic strategies in the treatment of fibrosis-related conjunctival diseases, based on the reversibility of both EMT process i epigenetic mechanisms.",
publisher = "Belgrade: Faculty of Biology, University of Belgrade",
journal = "Faculty of Biology, University of Belgrade",
title = "Uloga metilacije DNK u procesu epitelno-mezenhimske tranzicije  humanih epitelnih ćelija konjuktive, The role of DNA methylation in the process of epithelial-mesenchymal transition of human conjunctival epithelial cells",
pages = "1-133",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_3884"
}

Rajić, J.. (2020). Uloga metilacije DNK u procesu epitelno-mezenhimske tranzicije  humanih epitelnih ćelija konjuktive. in Faculty of Biology, University of Belgrade
Belgrade: Faculty of Biology, University of Belgrade., 1-133.
https://hdl.handle.net/21.15107/rcub_ibiss_3884

Rajić J. Uloga metilacije DNK u procesu epitelno-mezenhimske tranzicije  humanih epitelnih ćelija konjuktive. in Faculty of Biology, University of Belgrade. 2020;:1-133.
https://hdl.handle.net/21.15107/rcub_ibiss_3884 .

Rajić, Jovana, "Uloga metilacije DNK u procesu epitelno-mezenhimske tranzicije  humanih epitelnih ćelija konjuktive" in Faculty of Biology, University of Belgrade (2020):1-133,
https://hdl.handle.net/21.15107/rcub_ibiss_3884 .

TY  - CONF
AU  - Jakubek, Patrycja
AU  - Rajić, Jovana
AU  - Baranowska, Monika
AU  - Vidaković, Melita
AU  - Bartoszek, Agnieszka
AU  - Namiesnik, Jacek
PY  - 2019
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6319
AB  - The impact of catechins on the expression profile of redox-related genes in HT29 cell line has been studied recently by our group using Oxidative Stress RT2 Profiler PCR Array. Within the examined panel of 84 genes, the down-regulation of SRXN1 gene was unique among other-regulated genes. We hypothesized that the observed down-regulation resulted from DNA methylation and have exploited this observation to choose the proper strategy to monitor the changes in DNA methylation patterns incurred by dietary antioxidants. The current study verified two PCR-based approaches.
PB  - Basel: MDPI
C3  - CA16112—Luxemburg 2019 meeting; 2019 Mar 25-27; Luxemburg, Luxemburg
T1  - DNA Methylation Changes Induced by Redox-Active Compounds—Choosing the Right PCR-Based Method
IS  - 1
VL  - 11
DO  - 10.3390/proceedings2019011020
SP  - 20
ER  - 

@conference{
author = "Jakubek, Patrycja and Rajić, Jovana and Baranowska, Monika and Vidaković, Melita and Bartoszek, Agnieszka and Namiesnik, Jacek",
year = "2019",
abstract = "The impact of catechins on the expression profile of redox-related genes in HT29 cell line has been studied recently by our group using Oxidative Stress RT2 Profiler PCR Array. Within the examined panel of 84 genes, the down-regulation of SRXN1 gene was unique among other-regulated genes. We hypothesized that the observed down-regulation resulted from DNA methylation and have exploited this observation to choose the proper strategy to monitor the changes in DNA methylation patterns incurred by dietary antioxidants. The current study verified two PCR-based approaches.",
publisher = "Basel: MDPI",
journal = "CA16112—Luxemburg 2019 meeting; 2019 Mar 25-27; Luxemburg, Luxemburg",
title = "DNA Methylation Changes Induced by Redox-Active Compounds—Choosing the Right PCR-Based Method",
number = "1",
volume = "11",
doi = "10.3390/proceedings2019011020",
pages = "20"
}

Jakubek, P., Rajić, J., Baranowska, M., Vidaković, M., Bartoszek, A.,& Namiesnik, J.. (2019). DNA Methylation Changes Induced by Redox-Active Compounds—Choosing the Right PCR-Based Method. in CA16112—Luxemburg 2019 meeting; 2019 Mar 25-27; Luxemburg, Luxemburg
Basel: MDPI., 11(1), 20.
https://doi.org/10.3390/proceedings2019011020

Jakubek P, Rajić J, Baranowska M, Vidaković M, Bartoszek A, Namiesnik J. DNA Methylation Changes Induced by Redox-Active Compounds—Choosing the Right PCR-Based Method. in CA16112—Luxemburg 2019 meeting; 2019 Mar 25-27; Luxemburg, Luxemburg. 2019;11(1):20.
doi:10.3390/proceedings2019011020 .

Jakubek, Patrycja, Rajić, Jovana, Baranowska, Monika, Vidaković, Melita, Bartoszek, Agnieszka, Namiesnik, Jacek, "DNA Methylation Changes Induced by Redox-Active Compounds—Choosing the Right PCR-Based Method" in CA16112—Luxemburg 2019 meeting; 2019 Mar 25-27; Luxemburg, Luxemburg, 11, no. 1 (2019):20,
https://doi.org/10.3390/proceedings2019011020 . .

TY  - CONF
AU  - Jakubek, Patrycja
AU  - Baranowska, Monika
AU  - Rajić, Jovana
AU  - Vidaković, Melita
AU  - Namiesnik, Jacek
AU  - Bartoszek, Agnieszka
PY  - 2019
UR  - https://www.redox-medicine.com/images/2019/pdf/Paris_Redox_2019_-_Final_Agenda_-_V6.pdf
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6310
AB  - Introduction: Our previous research indicated that treatment of HT29 cells with different catechins affected the expression of redox-related genes in a dose dependent manner with only one gene (SRXN1) being down-regulated [1]. Since catechins were reported to affect DNA methylation levels [2], the objestive of current research was to find out wether the observed down-regulation of SRXN1 expression was caused by epigenetic changes.
Materials & Methods: Human colon adenocarcinoma HT29 cells were treated with 5 catechins at 4 concentrations for 24 hours and subsequently genomic DNA was isolated. To perform methylation analysis, DNA isolates were bisulfite converted using EZ DNA Methylation kit from Zymo Research (USA). DNA methylation profiles within the chosen fragment of promoter area of SRXN1 gene were examined using Methylation-Specific PCR and Methylation-Sensitive High Resolution Melting.
Results: According to Baranowska et al. [1], the treatment of HT29 cell line with 10 µM (-)-epigallocatechin caused down-regulation of SRXN1 gene. So, we hypothesized that the methylation level within the promoter CpG island of this gene will be increased by this compound and our presumption were confirmed. Besides, significant increase of DNA methylation was observed also for high dosed of (-)-epicatechin gallate.
Conclusion: Catechins may influence DNA methylation pattern of redox responsive genes.
PB  - Paris: University Pierre et Marie Curie
C3  - Abstracts book: 21st ISANH Interantional Conference on Oxidative stress, redox homeostasis and antioxidants; 2019 Jun 20-21; Paris, France
T1  - Modulation of DNA methylation profile of SRXN1 gene promoter in HT29 cells exposed to catechins of different redox activity
SP  - 42
EP  - 42
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6310
ER  - 

@conference{
author = "Jakubek, Patrycja and Baranowska, Monika and Rajić, Jovana and Vidaković, Melita and Namiesnik, Jacek and Bartoszek, Agnieszka",
year = "2019",
abstract = "Introduction: Our previous research indicated that treatment of HT29 cells with different catechins affected the expression of redox-related genes in a dose dependent manner with only one gene (SRXN1) being down-regulated [1]. Since catechins were reported to affect DNA methylation levels [2], the objestive of current research was to find out wether the observed down-regulation of SRXN1 expression was caused by epigenetic changes.
Materials & Methods: Human colon adenocarcinoma HT29 cells were treated with 5 catechins at 4 concentrations for 24 hours and subsequently genomic DNA was isolated. To perform methylation analysis, DNA isolates were bisulfite converted using EZ DNA Methylation kit from Zymo Research (USA). DNA methylation profiles within the chosen fragment of promoter area of SRXN1 gene were examined using Methylation-Specific PCR and Methylation-Sensitive High Resolution Melting.
Results: According to Baranowska et al. [1], the treatment of HT29 cell line with 10 µM (-)-epigallocatechin caused down-regulation of SRXN1 gene. So, we hypothesized that the methylation level within the promoter CpG island of this gene will be increased by this compound and our presumption were confirmed. Besides, significant increase of DNA methylation was observed also for high dosed of (-)-epicatechin gallate.
Conclusion: Catechins may influence DNA methylation pattern of redox responsive genes.",
publisher = "Paris: University Pierre et Marie Curie",
journal = "Abstracts book: 21st ISANH Interantional Conference on Oxidative stress, redox homeostasis and antioxidants; 2019 Jun 20-21; Paris, France",
title = "Modulation of DNA methylation profile of SRXN1 gene promoter in HT29 cells exposed to catechins of different redox activity",
pages = "42-42",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6310"
}

Jakubek, P., Baranowska, M., Rajić, J., Vidaković, M., Namiesnik, J.,& Bartoszek, A.. (2019). Modulation of DNA methylation profile of SRXN1 gene promoter in HT29 cells exposed to catechins of different redox activity. in Abstracts book: 21st ISANH Interantional Conference on Oxidative stress, redox homeostasis and antioxidants; 2019 Jun 20-21; Paris, France
Paris: University Pierre et Marie Curie., 42-42.
https://hdl.handle.net/21.15107/rcub_ibiss_6310

Jakubek P, Baranowska M, Rajić J, Vidaković M, Namiesnik J, Bartoszek A. Modulation of DNA methylation profile of SRXN1 gene promoter in HT29 cells exposed to catechins of different redox activity. in Abstracts book: 21st ISANH Interantional Conference on Oxidative stress, redox homeostasis and antioxidants; 2019 Jun 20-21; Paris, France. 2019;:42-42.
https://hdl.handle.net/21.15107/rcub_ibiss_6310 .

Jakubek, Patrycja, Baranowska, Monika, Rajić, Jovana, Vidaković, Melita, Namiesnik, Jacek, Bartoszek, Agnieszka, "Modulation of DNA methylation profile of SRXN1 gene promoter in HT29 cells exposed to catechins of different redox activity" in Abstracts book: 21st ISANH Interantional Conference on Oxidative stress, redox homeostasis and antioxidants; 2019 Jun 20-21; Paris, France (2019):42-42,
https://hdl.handle.net/21.15107/rcub_ibiss_6310 .

TY  - JOUR
AU  - Đorđević, Miloš
AU  - Grdović, Nevena
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Uskoković, Aleksandra
AU  - Rajić, Jovana
AU  - Đorđević, Marija
AU  - Tolić, Anja
AU  - Mišić, Danijela
AU  - Šiler, Branislav
AU  - Poznanović, Goran
AU  - Vidaković, Melita
AU  - Dinić, Svetlana
PY  - 2019
UR  - https://www.sciencedirect.com/science/article/pii/S0378874119315545?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3390
AB  - ETHNOPHARMACOLOGICAL RELEVANCE Centaurium erythraea Rafn (CE) is used as a traditional medicinal plant in Serbia to treat different ailments due to its antidiabetic, antipyretic, antiflatulent and detoxification effects. AIM OF THE STUDY Elucidation of the mechanisms that underlie the antioxidant and pro-survival effects of the CE extract (CEE) in beta-cells and pancreatic islets from streptozotocin (STZ)-treated diabetic rats. MATERIAL AND METHODS Diabetes was induced in rats by multiple applications of low doses of STZ (40 mg/kg intraperitoneally (i.p.), for five consecutive days). CEE (100 mg/kg) was administered orally, in the pre-treated group for two weeks before diabetes induction, during the treatments with STZ and for four weeks after diabetes onset, and in the post-treatment group for four weeks after diabetes induction. The impact of CEE on diabetic islets was estimated by histological and immunohistochemical examination of the pancreas. Molecular mechanisms of the effects of CEE were also analyzed in insulinoma Rin-5F cells treated with STZ (12 mM) and CEE (0.25 mg/mL). Oxidative stress was evaluated by assessing the levels of DNA damage, lipid peroxidation, protein S-glutathionylation and enzymatic activities and expression of CAT, MnSOD, CuZnSOD, GPx and GR in beta-cells. The presence and activities of the redox-sensitive and islet-enriched regulatory proteins were also analyzed. RESULTS Treatment with CEE ameliorated the insulin level and glycemic control in STZ-induced diabetic rats by improving the structural and functional properties of pancreatic islets through multiple routes of action. The disturbance of islet morphology and islet cell contents in diabetes was reduced by the CEE treatment and was associated with a protective effect of CEE on the levels of insulin, GLUT-2 and p-Akt in diabetic islets. The antioxidant effect of CEE on STZ-treated beta-cells was displayed as reduced DNA damage, lipid peroxidation, protein S-glutathionylation and alleviation of STZ-induced disruption in MnSOD, CuZnSOD and CAT enzyme activities. The oxidative stress-induced disturbance of the transcriptional regulation of CAT, MnSOD, CuZnSOD, GPx and GR enzymes in beta-cells was improved after the CEE treatment, and was observed as readjustment of the presence and activities of redox-sensitive NFκB-p65, FOXO3A, Sp1 and Nrf-2 transcription factors. The observed CEE-mediated induction of proliferative and pro-survival pathways and insulin expression/secretion after STZ-induced oxidative stress in beta-cells could be partially attributed to a fine-tuned modulation of the activities of pro-survival Akt, ERK and p38 kinases and islet-enriched Pdx-1 and MafA regulatory factors. CONCLUSIONS The results of this study provide evidence that CEE improves the structural and functional properties of pancreatic beta-cells by correcting the endogenous antioxidant regulatory mechanisms and by promoting proliferative and pro-survival pathways in beta-cells.
T2  - Journal of Ethnopharmacology
T1  - Centaurium erythraea extract improves survival and functionality of pancreatic beta-cells in diabetes through multiple routes of action.
VL  - 242
DO  - 10.1016/j.jep.2019.112043
SP  - 112043
ER  - 

@article{
author = "Đorđević, Miloš and Grdović, Nevena and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Uskoković, Aleksandra and Rajić, Jovana and Đorđević, Marija and Tolić, Anja and Mišić, Danijela and Šiler, Branislav and Poznanović, Goran and Vidaković, Melita and Dinić, Svetlana",
year = "2019",
abstract = "ETHNOPHARMACOLOGICAL RELEVANCE Centaurium erythraea Rafn (CE) is used as a traditional medicinal plant in Serbia to treat different ailments due to its antidiabetic, antipyretic, antiflatulent and detoxification effects. AIM OF THE STUDY Elucidation of the mechanisms that underlie the antioxidant and pro-survival effects of the CE extract (CEE) in beta-cells and pancreatic islets from streptozotocin (STZ)-treated diabetic rats. MATERIAL AND METHODS Diabetes was induced in rats by multiple applications of low doses of STZ (40 mg/kg intraperitoneally (i.p.), for five consecutive days). CEE (100 mg/kg) was administered orally, in the pre-treated group for two weeks before diabetes induction, during the treatments with STZ and for four weeks after diabetes onset, and in the post-treatment group for four weeks after diabetes induction. The impact of CEE on diabetic islets was estimated by histological and immunohistochemical examination of the pancreas. Molecular mechanisms of the effects of CEE were also analyzed in insulinoma Rin-5F cells treated with STZ (12 mM) and CEE (0.25 mg/mL). Oxidative stress was evaluated by assessing the levels of DNA damage, lipid peroxidation, protein S-glutathionylation and enzymatic activities and expression of CAT, MnSOD, CuZnSOD, GPx and GR in beta-cells. The presence and activities of the redox-sensitive and islet-enriched regulatory proteins were also analyzed. RESULTS Treatment with CEE ameliorated the insulin level and glycemic control in STZ-induced diabetic rats by improving the structural and functional properties of pancreatic islets through multiple routes of action. The disturbance of islet morphology and islet cell contents in diabetes was reduced by the CEE treatment and was associated with a protective effect of CEE on the levels of insulin, GLUT-2 and p-Akt in diabetic islets. The antioxidant effect of CEE on STZ-treated beta-cells was displayed as reduced DNA damage, lipid peroxidation, protein S-glutathionylation and alleviation of STZ-induced disruption in MnSOD, CuZnSOD and CAT enzyme activities. The oxidative stress-induced disturbance of the transcriptional regulation of CAT, MnSOD, CuZnSOD, GPx and GR enzymes in beta-cells was improved after the CEE treatment, and was observed as readjustment of the presence and activities of redox-sensitive NFκB-p65, FOXO3A, Sp1 and Nrf-2 transcription factors. The observed CEE-mediated induction of proliferative and pro-survival pathways and insulin expression/secretion after STZ-induced oxidative stress in beta-cells could be partially attributed to a fine-tuned modulation of the activities of pro-survival Akt, ERK and p38 kinases and islet-enriched Pdx-1 and MafA regulatory factors. CONCLUSIONS The results of this study provide evidence that CEE improves the structural and functional properties of pancreatic beta-cells by correcting the endogenous antioxidant regulatory mechanisms and by promoting proliferative and pro-survival pathways in beta-cells.",
journal = "Journal of Ethnopharmacology",
title = "Centaurium erythraea extract improves survival and functionality of pancreatic beta-cells in diabetes through multiple routes of action.",
volume = "242",
doi = "10.1016/j.jep.2019.112043",
pages = "112043"
}

Đorđević, M., Grdović, N., Mihailović, M., Arambašić Jovanović, J., Uskoković, A., Rajić, J., Đorđević, M., Tolić, A., Mišić, D., Šiler, B., Poznanović, G., Vidaković, M.,& Dinić, S.. (2019). Centaurium erythraea extract improves survival and functionality of pancreatic beta-cells in diabetes through multiple routes of action.. in Journal of Ethnopharmacology, 242, 112043.
https://doi.org/10.1016/j.jep.2019.112043

Đorđević M, Grdović N, Mihailović M, Arambašić Jovanović J, Uskoković A, Rajić J, Đorđević M, Tolić A, Mišić D, Šiler B, Poznanović G, Vidaković M, Dinić S. Centaurium erythraea extract improves survival and functionality of pancreatic beta-cells in diabetes through multiple routes of action.. in Journal of Ethnopharmacology. 2019;242:112043.
doi:10.1016/j.jep.2019.112043 .

Đorđević, Miloš, Grdović, Nevena, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Uskoković, Aleksandra, Rajić, Jovana, Đorđević, Marija, Tolić, Anja, Mišić, Danijela, Šiler, Branislav, Poznanović, Goran, Vidaković, Melita, Dinić, Svetlana, "Centaurium erythraea extract improves survival and functionality of pancreatic beta-cells in diabetes through multiple routes of action." in Journal of Ethnopharmacology, 242 (2019):112043,
https://doi.org/10.1016/j.jep.2019.112043 . .