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  - CONF
AU  - Rajić, Jovana
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Mihailović, Mirjana
AU  - Dinić, Svetlana
AU  - Uskoković, Aleksandra
AU  - Arambašić Jovanović, Jelena
AU  - Poznanović, Goran
AU  - Inić-Kanada, Aleksandra
AU  - Barisani-Asenbauer, Talin
AU  - Grdović, Nevena
AU  - Vidaković, Melita
PY  - 2019
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5697
AB  - Conjunctival fibrosis often emerges after infections, inflammations and mechanical stresses of the eye and when severe results in impaired vision. Recent data documented unavoidable role of epithelial to mesenchymal transition (EMT) in every fibrotic process including fibrosis-based eye conditions. The aim of this work was to establish whether human conjunctival epithelial (HCjE) cells are prone to EMT induction after prolonged treatment with well-known EMT inducers TGF-β proteins, and to test capabilities of TGF-β1, TGF-β2 and their combination to trigger this process. While TGF-β2 induced only alterations in cell-cell adhesion, TGF-β1 and combination of TGF-β proteins induced prominent change in cell morphology reflected in loss of cell-cell contacts, changes in shape from epithelial polygonal to spindle-like shape typical for mesenchymal phenotype and acquired ability to move. Statistically significant reduction of mRNA expression of epithelial marker genes (CDH1, OCLN, DSP) was observed in all treatment groups, while mRNA expression level of mesenchymal marker genes (CDH2, FN1, VIM) and EMT-related transcription factors (SNAI1, ZEB2, TWIST1) varied among treatment groups. TGF-β1 treatment induced the most pronounced increase in the level of mRNA of genes characteristic for mesenchymal phenotype that was accompanied with corresponding increase in protein level of two mesenchymal markers (CDH2, FN1), in parallel with decrease in protein expression of two epithelial markers (CDH1, DSP). To conclude, HCjE cells are prone to EMT induction and TGF-β1 possesses the highest potential for EMT induction in HCjE cells, suggesting that, in conditions of chronic inflammation, induction of EMT in conjunctival cells could contribute to fibrosis-related eye diseases.
PB  - Belgrade: Faculty of Chemistry
C3  - The 9th Conference of the Serbian Biochemical Society: Diversity in Biochemistry; 2019 Nov 14-16; Belgrade, Serbia
T1  - The capability of different TGF-β isoforms to induce EMT in human conjunctival epithelial cells
SP  - 159
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5697
ER  - 

@conference{
author = "Rajić, Jovana and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Mihailović, Mirjana and Dinić, Svetlana and Uskoković, Aleksandra and Arambašić Jovanović, Jelena and Poznanović, Goran and Inić-Kanada, Aleksandra and Barisani-Asenbauer, Talin and Grdović, Nevena and Vidaković, Melita",
year = "2019",
abstract = "Conjunctival fibrosis often emerges after infections, inflammations and mechanical stresses of the eye and when severe results in impaired vision. Recent data documented unavoidable role of epithelial to mesenchymal transition (EMT) in every fibrotic process including fibrosis-based eye conditions. The aim of this work was to establish whether human conjunctival epithelial (HCjE) cells are prone to EMT induction after prolonged treatment with well-known EMT inducers TGF-β proteins, and to test capabilities of TGF-β1, TGF-β2 and their combination to trigger this process. While TGF-β2 induced only alterations in cell-cell adhesion, TGF-β1 and combination of TGF-β proteins induced prominent change in cell morphology reflected in loss of cell-cell contacts, changes in shape from epithelial polygonal to spindle-like shape typical for mesenchymal phenotype and acquired ability to move. Statistically significant reduction of mRNA expression of epithelial marker genes (CDH1, OCLN, DSP) was observed in all treatment groups, while mRNA expression level of mesenchymal marker genes (CDH2, FN1, VIM) and EMT-related transcription factors (SNAI1, ZEB2, TWIST1) varied among treatment groups. TGF-β1 treatment induced the most pronounced increase in the level of mRNA of genes characteristic for mesenchymal phenotype that was accompanied with corresponding increase in protein level of two mesenchymal markers (CDH2, FN1), in parallel with decrease in protein expression of two epithelial markers (CDH1, DSP). To conclude, HCjE cells are prone to EMT induction and TGF-β1 possesses the highest potential for EMT induction in HCjE cells, suggesting that, in conditions of chronic inflammation, induction of EMT in conjunctival cells could contribute to fibrosis-related eye diseases.",
publisher = "Belgrade: Faculty of Chemistry",
journal = "The 9th Conference of the Serbian Biochemical Society: Diversity in Biochemistry; 2019 Nov 14-16; Belgrade, Serbia",
title = "The capability of different TGF-β isoforms to induce EMT in human conjunctival epithelial cells",
pages = "159",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5697"
}

Rajić, J., Tolić, A., Đorđević, M., Đorđević, M., Mihailović, M., Dinić, S., Uskoković, A., Arambašić Jovanović, J., Poznanović, G., Inić-Kanada, A., Barisani-Asenbauer, T., Grdović, N.,& Vidaković, M.. (2019). The capability of different TGF-β isoforms to induce EMT in human conjunctival epithelial cells. in The 9th Conference of the Serbian Biochemical Society: Diversity in Biochemistry; 2019 Nov 14-16; Belgrade, Serbia
Belgrade: Faculty of Chemistry., 159.
https://hdl.handle.net/21.15107/rcub_ibiss_5697

Rajić J, Tolić A, Đorđević M, Đorđević M, Mihailović M, Dinić S, Uskoković A, Arambašić Jovanović J, Poznanović G, Inić-Kanada A, Barisani-Asenbauer T, Grdović N, Vidaković M. The capability of different TGF-β isoforms to induce EMT in human conjunctival epithelial cells. in The 9th Conference of the Serbian Biochemical Society: Diversity in Biochemistry; 2019 Nov 14-16; Belgrade, Serbia. 2019;:159.
https://hdl.handle.net/21.15107/rcub_ibiss_5697 .

Rajić, Jovana, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Mihailović, Mirjana, Dinić, Svetlana, Uskoković, Aleksandra, Arambašić Jovanović, Jelena, Poznanović, Goran, Inić-Kanada, Aleksandra, Barisani-Asenbauer, Talin, Grdović, Nevena, Vidaković, Melita, "The capability of different TGF-β isoforms to induce EMT in human conjunctival epithelial cells" in The 9th Conference of the Serbian Biochemical Society: Diversity in Biochemistry; 2019 Nov 14-16; Belgrade, Serbia (2019):159,
https://hdl.handle.net/21.15107/rcub_ibiss_5697 .

TY  - CONF
AU  - Ghasemian, Ehsan
AU  - Stein, Elisabeth
AU  - Inić-Kanada, Aleksandra
AU  - Schuerer, Nadine
AU  - Bintner, Nora
AU  - Rajić, Jovana
AU  - Barisani-Asenbauer, Talin
PY  - 2017
UR  - https://iovs.arvojournals.org/article.aspx?articleid=2642535
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6313
AB  - Purpose : Bacterial ghosts (BGs) are empty cell envelopes derived from Gram-negative bacteria. BGs possess all cell membrane structures, like live bacteria they are originated from, which gives a potential to use them as an adjuvant delivery system. In the present study, we investigated the potential of Escherichia coli Nissle (EcN) and Salmonella typhimurium (Sty) BGs in modulation of immune responses in human conjunctival epithelial cells (HCjE).
Methods : Confluent monolayers of HCjE cells were treated for 2 and 24 hours with EcN and Sty BGs at a concentration of 1x108 particles/ml, as well as with Interleukin- 1α as a general stimulator. 24 hours after cell stimulation the supernatants and cells were collected. Concentration of IL-6 and IL-8 were measured using ELISA. Quantification of the gene expression levels for TLR-4 and TLR-5 was performed using qRT-PCR.
Results : HCjE cells responded to EcN BGs and Sty BGs with increased secretion of pro-inflammatory cytokines and expression of toll-like receptors (TLRs). Expression of TLR-4 and TLR-5 was significantly higher in EcN BGs stimulated cells compared to Sty BGs stimulated HCjE cells (P<0.05). EcN and Sty BGs induced enhanced secretion of IL-6 and IL-8 in HCjE cells.
Conclusions : Our results suggest that both used BGs may modulate the secretion of IL-6 and IL-8 mainly via the activation of TLR-4 and TLR-5 in HCjE cells.
PB  - Rockville: Association for Research in Vision and Ophthalmolog Inc.
C3  - Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA
T1  - The effects of Escherichia coli Nissle and Salmonella typhimurium bacterial ghosts on human conjunctival epithelial cells
SP  - 5773
EP  - 5773
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6313
ER  - 

@conference{
author = "Ghasemian, Ehsan and Stein, Elisabeth and Inić-Kanada, Aleksandra and Schuerer, Nadine and Bintner, Nora and Rajić, Jovana and Barisani-Asenbauer, Talin",
year = "2017",
abstract = "Purpose : Bacterial ghosts (BGs) are empty cell envelopes derived from Gram-negative bacteria. BGs possess all cell membrane structures, like live bacteria they are originated from, which gives a potential to use them as an adjuvant delivery system. In the present study, we investigated the potential of Escherichia coli Nissle (EcN) and Salmonella typhimurium (Sty) BGs in modulation of immune responses in human conjunctival epithelial cells (HCjE).
Methods : Confluent monolayers of HCjE cells were treated for 2 and 24 hours with EcN and Sty BGs at a concentration of 1x108 particles/ml, as well as with Interleukin- 1α as a general stimulator. 24 hours after cell stimulation the supernatants and cells were collected. Concentration of IL-6 and IL-8 were measured using ELISA. Quantification of the gene expression levels for TLR-4 and TLR-5 was performed using qRT-PCR.
Results : HCjE cells responded to EcN BGs and Sty BGs with increased secretion of pro-inflammatory cytokines and expression of toll-like receptors (TLRs). Expression of TLR-4 and TLR-5 was significantly higher in EcN BGs stimulated cells compared to Sty BGs stimulated HCjE cells (P<0.05). EcN and Sty BGs induced enhanced secretion of IL-6 and IL-8 in HCjE cells.
Conclusions : Our results suggest that both used BGs may modulate the secretion of IL-6 and IL-8 mainly via the activation of TLR-4 and TLR-5 in HCjE cells.",
publisher = "Rockville: Association for Research in Vision and Ophthalmolog Inc.",
journal = "Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA",
title = "The effects of Escherichia coli Nissle and Salmonella typhimurium bacterial ghosts on human conjunctival epithelial cells",
pages = "5773-5773",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6313"
}

Ghasemian, E., Stein, E., Inić-Kanada, A., Schuerer, N., Bintner, N., Rajić, J.,& Barisani-Asenbauer, T.. (2017). The effects of Escherichia coli Nissle and Salmonella typhimurium bacterial ghosts on human conjunctival epithelial cells. in Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA
Rockville: Association for Research in Vision and Ophthalmolog Inc.., 5773-5773.
https://hdl.handle.net/21.15107/rcub_ibiss_6313

Ghasemian E, Stein E, Inić-Kanada A, Schuerer N, Bintner N, Rajić J, Barisani-Asenbauer T. The effects of Escherichia coli Nissle and Salmonella typhimurium bacterial ghosts on human conjunctival epithelial cells. in Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA. 2017;:5773-5773.
https://hdl.handle.net/21.15107/rcub_ibiss_6313 .

Ghasemian, Ehsan, Stein, Elisabeth, Inić-Kanada, Aleksandra, Schuerer, Nadine, Bintner, Nora, Rajić, Jovana, Barisani-Asenbauer, Talin, "The effects of Escherichia coli Nissle and Salmonella typhimurium bacterial ghosts on human conjunctival epithelial cells" in Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA (2017):5773-5773,
https://hdl.handle.net/21.15107/rcub_ibiss_6313 .

TY  - CONF
AU  - Barisani-Asenbauer, Talin
AU  - Inić-Kanada, Aleksandra
AU  - Rajić, Jovana
AU  - Grdović, Nevena
AU  - Stein, Elisabeth
AU  - Ghasemian, Ehsan
AU  - Schuerer, Nadine
AU  - Vidaković, Melita
PY  - 2017
UR  - https://iovs.arvojournals.org/article.aspx?articleid=2642540
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6314
AB  - Purpose : Epithelial-mesenchymal transition (EMT) of conjunctival epithelium may contribute to trachoma-associated conjunctival fibrosis. Epigenetic mechanisms have an important role in different disease etiologies, however, it is unknown whether they play a role during Chlamydia trachomatis (Ct) ocular infection. The aim of this study was to test the ability of Ct to induce EMT in human conjunctival epithelial (HCjE) cells in vitro and investigate whether alterations in gene expression after Ct infection are correlated with DNA methylation of EMT related marker genes.
Methods : mRNA and protein expression levels of EMT markers, including TGFβ1 and TGFβ2, E-cadherin (CDH1), fibronectin (FN1) and α-smooth muscle actin (ACTA2) were assayed by qPCR and Western blot analysis in HCjE cells infected with Ct serovar B. Promoters and gene-bodies of CDH1, FN1 and ACTA2 were analyzed for the presence of CpG islands using CpG Island Searcher software and thereafter changes in DNA methylation patterns of selected regions were examined with MSP, HRM and bisulfite sequencing.
Results : Infection of HCjE cells with Ct resulted in a statistically significant increase in TGFβ1 and TGFβ2 mRNA and significant reduction of mRNA expression of E-cadherin gene (CDH1), wherein CDH1 mRNA expression was 2.7-fold higher in a control than in Ct infected HCjE cells. Decrease in mRNA expression was accompanied with diminishment in protein level of E-cadherin after Ct infection. HRM data showed convincing difference between control and Ct infected level of DNA methylation with increase from 12.8% to 21.8% suggesting that CDH1 promoter region is subjected to alterations in DNA methylation in HCjE cells because of Ct infection.
Conclusions : Our data for the first time indicate a direct role of Ct infection in epigenetic regulation of human conjunctival epithelium. Studying the pathways of EMT in HCjE cells may help to develop new therapeutic strategies to treat conjunctival fibrosis.
PB  - Rockville: Association for Research in Vision and Ophthalmolog Inc.
C3  - Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA
T1  - Chlamydia trachomatis infection induces epithelial-mesenchymal transition in conjunctival epithelial cells
SP  - 5778
EP  - 5778
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6314
ER  - 

@conference{
author = "Barisani-Asenbauer, Talin and Inić-Kanada, Aleksandra and Rajić, Jovana and Grdović, Nevena and Stein, Elisabeth and Ghasemian, Ehsan and Schuerer, Nadine and Vidaković, Melita",
year = "2017",
abstract = "Purpose : Epithelial-mesenchymal transition (EMT) of conjunctival epithelium may contribute to trachoma-associated conjunctival fibrosis. Epigenetic mechanisms have an important role in different disease etiologies, however, it is unknown whether they play a role during Chlamydia trachomatis (Ct) ocular infection. The aim of this study was to test the ability of Ct to induce EMT in human conjunctival epithelial (HCjE) cells in vitro and investigate whether alterations in gene expression after Ct infection are correlated with DNA methylation of EMT related marker genes.
Methods : mRNA and protein expression levels of EMT markers, including TGFβ1 and TGFβ2, E-cadherin (CDH1), fibronectin (FN1) and α-smooth muscle actin (ACTA2) were assayed by qPCR and Western blot analysis in HCjE cells infected with Ct serovar B. Promoters and gene-bodies of CDH1, FN1 and ACTA2 were analyzed for the presence of CpG islands using CpG Island Searcher software and thereafter changes in DNA methylation patterns of selected regions were examined with MSP, HRM and bisulfite sequencing.
Results : Infection of HCjE cells with Ct resulted in a statistically significant increase in TGFβ1 and TGFβ2 mRNA and significant reduction of mRNA expression of E-cadherin gene (CDH1), wherein CDH1 mRNA expression was 2.7-fold higher in a control than in Ct infected HCjE cells. Decrease in mRNA expression was accompanied with diminishment in protein level of E-cadherin after Ct infection. HRM data showed convincing difference between control and Ct infected level of DNA methylation with increase from 12.8% to 21.8% suggesting that CDH1 promoter region is subjected to alterations in DNA methylation in HCjE cells because of Ct infection.
Conclusions : Our data for the first time indicate a direct role of Ct infection in epigenetic regulation of human conjunctival epithelium. Studying the pathways of EMT in HCjE cells may help to develop new therapeutic strategies to treat conjunctival fibrosis.",
publisher = "Rockville: Association for Research in Vision and Ophthalmolog Inc.",
journal = "Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA",
title = "Chlamydia trachomatis infection induces epithelial-mesenchymal transition in conjunctival epithelial cells",
pages = "5778-5778",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6314"
}

Barisani-Asenbauer, T., Inić-Kanada, A., Rajić, J., Grdović, N., Stein, E., Ghasemian, E., Schuerer, N.,& Vidaković, M.. (2017). Chlamydia trachomatis infection induces epithelial-mesenchymal transition in conjunctival epithelial cells. in Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA
Rockville: Association for Research in Vision and Ophthalmolog Inc.., 5778-5778.
https://hdl.handle.net/21.15107/rcub_ibiss_6314

Barisani-Asenbauer T, Inić-Kanada A, Rajić J, Grdović N, Stein E, Ghasemian E, Schuerer N, Vidaković M. Chlamydia trachomatis infection induces epithelial-mesenchymal transition in conjunctival epithelial cells. in Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA. 2017;:5778-5778.
https://hdl.handle.net/21.15107/rcub_ibiss_6314 .

Barisani-Asenbauer, Talin, Inić-Kanada, Aleksandra, Rajić, Jovana, Grdović, Nevena, Stein, Elisabeth, Ghasemian, Ehsan, Schuerer, Nadine, Vidaković, Melita, "Chlamydia trachomatis infection induces epithelial-mesenchymal transition in conjunctival epithelial cells" in Annual Meeting of the Association-for-Research-in-Vision-and-Ophthalmology (ARVO); 2017 May 7-11; Baltimore, USA (2017):5778-5778,
https://hdl.handle.net/21.15107/rcub_ibiss_6314 .

TY  - CONF
AU  - Rajić, Jovana
AU  - Grdović, Nevena
AU  - Inic-Kanada, Aleksandra
AU  - Stein, Elisabeth
AU  - Schuerer, Nadine
AU  - Uskoković, Aleksandra
AU  - Ghasemian, Ehsan
AU  - Dinić, Svetlana
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Poznanović, Goran
AU  - Barisani-Asenbauer, Talin
AU  - Vidaković, Melita
PY  - 2017
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2866
UR  - http://molbios.bio.bg.ac.rs/combos
AB  - Trachoma is the leading infectious cause of blindness worldwide initiated by repeated infections of the conjunctiva with Chlamydia trachomatis (Ct). Consequent chronic inflammation results in formation of fibrotic tissue which can eventually lead to damage of the corneal surface. Lately, emerging evidence suggests involvement of epithelial to mesenchymaltransition (EMT) in development of every fibrotic process. Process of EMT represents reversible transition of epithelial into mesenchymalcells that can be traced through numerous markers such as E-cadherin, highly expressed in epithelial cells, α-SMA, a frequently used mesenchymalEMT marker and fibronectin, a glycoprotein whose expression increases during the EMT process. Recent findings have suggested that epigenetic events are master regulators of expression of many EMT related genes.
C3  - 1st Congress of Molecular Biologists of Serbia
T1  - Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva: possible epigenetic mechanisms unveiled
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_2866
ER  - 

@conference{
author = "Rajić, Jovana and Grdović, Nevena and Inic-Kanada, Aleksandra and Stein, Elisabeth and Schuerer, Nadine and Uskoković, Aleksandra and Ghasemian, Ehsan and Dinić, Svetlana and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Poznanović, Goran and Barisani-Asenbauer, Talin and Vidaković, Melita",
year = "2017",
abstract = "Trachoma is the leading infectious cause of blindness worldwide initiated by repeated infections of the conjunctiva with Chlamydia trachomatis (Ct). Consequent chronic inflammation results in formation of fibrotic tissue which can eventually lead to damage of the corneal surface. Lately, emerging evidence suggests involvement of epithelial to mesenchymaltransition (EMT) in development of every fibrotic process. Process of EMT represents reversible transition of epithelial into mesenchymalcells that can be traced through numerous markers such as E-cadherin, highly expressed in epithelial cells, α-SMA, a frequently used mesenchymalEMT marker and fibronectin, a glycoprotein whose expression increases during the EMT process. Recent findings have suggested that epigenetic events are master regulators of expression of many EMT related genes.",
journal = "1st Congress of Molecular Biologists of Serbia",
title = "Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva: possible epigenetic mechanisms unveiled",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_2866"
}

Rajić, J., Grdović, N., Inic-Kanada, A., Stein, E., Schuerer, N., Uskoković, A., Ghasemian, E., Dinić, S., Mihailović, M., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Đorđević, M., Poznanović, G., Barisani-Asenbauer, T.,& Vidaković, M.. (2017). Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva: possible epigenetic mechanisms unveiled. in 1st Congress of Molecular Biologists of Serbia.
https://hdl.handle.net/21.15107/rcub_ibiss_2866

Rajić J, Grdović N, Inic-Kanada A, Stein E, Schuerer N, Uskoković A, Ghasemian E, Dinić S, Mihailović M, Arambašić Jovanović J, Tolić A, Đorđević M, Đorđević M, Poznanović G, Barisani-Asenbauer T, Vidaković M. Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva: possible epigenetic mechanisms unveiled. in 1st Congress of Molecular Biologists of Serbia. 2017;.
https://hdl.handle.net/21.15107/rcub_ibiss_2866 .

Rajić, Jovana, Grdović, Nevena, Inic-Kanada, Aleksandra, Stein, Elisabeth, Schuerer, Nadine, Uskoković, Aleksandra, Ghasemian, Ehsan, Dinić, Svetlana, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Poznanović, Goran, Barisani-Asenbauer, Talin, Vidaković, Melita, "Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva: possible epigenetic mechanisms unveiled" in 1st Congress of Molecular Biologists of Serbia (2017),
https://hdl.handle.net/21.15107/rcub_ibiss_2866 .

TY  - CONF
AU  - Rajić, Jovana
AU  - Grdović, Nevena
AU  - Inic-Kanada, Aleksandra
AU  - Stein, Elisabeth
AU  - Schuerer, Nadine
AU  - Uskoković, Aleksandra
AU  - Ghasemian, Ehsan
AU  - Dinić, Svetlana
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Poznanović, Goran
AU  - Barisani-Asenbauer, Talin
AU  - Vidaković, Melita
PY  - 2017
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2869
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2866
AB  - Introduction: Trachoma is the most common cause of infectious blindness worldwide, initiated by repeated infection of the conjunctiva with Chlamydia trachomatis (Ct). The resulting chronic inflammation and formation of fibrotic tissue eventually lead to corneal damage. Based on the facts that epithelial to mesenchymal transition (EMT) plays an important role in the development of fibrosis and that EMT is epigenetically regulated process, the aims of this study were to reveal the capacity of Ct to induce EMT in vitro and to unveil potential underlying epigenetic mechanisms. Methods: Human conjunctival epithelial (HCjE) cells were infected with 107 IFU of Ct for 72 h. EMT-inducing signaling pathways, as well as mRNA and protein expression of EMT markers (E-cadherin, fibronectin and α-SMA) were evaluated by RT-qPCR, Immunoblotting and Immunocytochemistry. DNA methylation patterns of selected regions of E-cadherin, fibronectin and α-SMA genes were examined by Methylation-Specific PCR, High Resolution Melting analysis and Bisulfite Sequencing. Results: Infection with Ct was accompanied with the activation of EMT-inducing signaling pathways, downregulation of epithelial marker E-cadherin and upregulation of mesenchymal markers fibronectin and α-SMA. While DNA methylation status of E-cadherin gene promoter correlated with its expression, methylation status of fibronectin and α-SMA genes couldn’t be related to their expression levels. Conclusion: Ct infection of HCjE cells triggers EMT that goes along with changes in the methylation profile of the E-cadherin promoter. Sequence of events described herein could contribute to scarring process in trachoma and open up possibilities for development of new therapeutic strategies in trachoma treatment.
PB  - Belgrade: University of Belgrade, Faculty of Biology
C3  - 1st Congress of Molecular Biologists of Serbia
T1  - Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled
SP  - 70
EP  - 70
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_2869
ER  - 

@conference{
editor = "Brajušković, Goran, Đorđević, Ana",
author = "Rajić, Jovana and Grdović, Nevena and Inic-Kanada, Aleksandra and Stein, Elisabeth and Schuerer, Nadine and Uskoković, Aleksandra and Ghasemian, Ehsan and Dinić, Svetlana and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Poznanović, Goran and Barisani-Asenbauer, Talin and Vidaković, Melita",
year = "2017",
abstract = "Introduction: Trachoma is the most common cause of infectious blindness worldwide, initiated by repeated infection of the conjunctiva with Chlamydia trachomatis (Ct). The resulting chronic inflammation and formation of fibrotic tissue eventually lead to corneal damage. Based on the facts that epithelial to mesenchymal transition (EMT) plays an important role in the development of fibrosis and that EMT is epigenetically regulated process, the aims of this study were to reveal the capacity of Ct to induce EMT in vitro and to unveil potential underlying epigenetic mechanisms. Methods: Human conjunctival epithelial (HCjE) cells were infected with 107 IFU of Ct for 72 h. EMT-inducing signaling pathways, as well as mRNA and protein expression of EMT markers (E-cadherin, fibronectin and α-SMA) were evaluated by RT-qPCR, Immunoblotting and Immunocytochemistry. DNA methylation patterns of selected regions of E-cadherin, fibronectin and α-SMA genes were examined by Methylation-Specific PCR, High Resolution Melting analysis and Bisulfite Sequencing. Results: Infection with Ct was accompanied with the activation of EMT-inducing signaling pathways, downregulation of epithelial marker E-cadherin and upregulation of mesenchymal markers fibronectin and α-SMA. While DNA methylation status of E-cadherin gene promoter correlated with its expression, methylation status of fibronectin and α-SMA genes couldn’t be related to their expression levels. Conclusion: Ct infection of HCjE cells triggers EMT that goes along with changes in the methylation profile of the E-cadherin promoter. Sequence of events described herein could contribute to scarring process in trachoma and open up possibilities for development of new therapeutic strategies in trachoma treatment.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology",
journal = "1st Congress of Molecular Biologists of Serbia",
title = "Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled",
pages = "70-70",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_2869"
}

Brajušković, G., Đorđević, A., Rajić, J., Grdović, N., Inic-Kanada, A., Stein, E., Schuerer, N., Uskoković, A., Ghasemian, E., Dinić, S., Mihailović, M., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Đorđević, M., Poznanović, G., Barisani-Asenbauer, T.,& Vidaković, M.. (2017). Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled. in 1st Congress of Molecular Biologists of Serbia
Belgrade: University of Belgrade, Faculty of Biology., 70-70.
https://hdl.handle.net/21.15107/rcub_ibiss_2869

Brajušković G, Đorđević A, Rajić J, Grdović N, Inic-Kanada A, Stein E, Schuerer N, Uskoković A, Ghasemian E, Dinić S, Mihailović M, Arambašić Jovanović J, Tolić A, Đorđević M, Đorđević M, Poznanović G, Barisani-Asenbauer T, Vidaković M. Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled. in 1st Congress of Molecular Biologists of Serbia. 2017;:70-70.
https://hdl.handle.net/21.15107/rcub_ibiss_2869 .

Brajušković, Goran, Đorđević, Ana, Rajić, Jovana, Grdović, Nevena, Inic-Kanada, Aleksandra, Stein, Elisabeth, Schuerer, Nadine, Uskoković, Aleksandra, Ghasemian, Ehsan, Dinić, Svetlana, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Poznanović, Goran, Barisani-Asenbauer, Talin, Vidaković, Melita, "Chlamydia trachomatis infection and development of epithelial mesenchymal transition in conjunctiva : possible epigenetic mechanisms unveiled" in 1st Congress of Molecular Biologists of Serbia (2017):70-70,
https://hdl.handle.net/21.15107/rcub_ibiss_2869 .

TY  - CONF
AU  - Rajić, Jovana
AU  - Grdović, Nevena
AU  - Inić-Kanada, Aleksandra
AU  - Stein, Elisabath
AU  - Schuerer, Nadine
AU  - Uskoković, Aleksandra
AU  - Ghasemian, Ehsan
AU  - Dinić, Svetlana
AU  - Mihailović, Mirjana
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Marija
AU  - Đorđević, Miloš
AU  - Poznanović, Goran
AU  - Barisani-Asenbauer, Talin
AU  - Vidaković, Melita
PY  - 2017
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5814
AB  - Repeated eye infections with Chlamydia trachomatis (Ct) usually lead to conjunctiva! scarring, corneal damage and blindness (trachoma). Considering epithelial to mesenchymal transition (EMD as unavoidable player in the development of fibrosis and taking into account that EMT is epigenetically reg­ulated process, our intention was to explore whether Ct infec­tion induces EMT in vitro and to uncover potential underlying epigenetic mechanisms. 
Human conjunctiva! epithelial cells were infected with 107 IFU of Ct for 72 h. The mRNA and protein expression of EMT mark­ers (E-cadherin and fibronectin) were assessed by RT-qPCR, lmmunoblotting and lmmunocytochemfstry. DNA methyfation analysis of selected regions of marker genes were examined by MSP, HRM and Bs-Seq. 
The Ct infection induced EMT-related changes in mRNA and protein expression, downregulation of epithelial marker E-cadherin and upregulation of mesenchymal marker fibronectin. Increase in DNA methylation of E-cadherin gene promoter was in correlation with its decreased expression, while DNA meth­ylation status of fibronectin gene couldn't be related to its ex­pression level. 
We suggest that manipulating the EMT process via modula­tion of DNA methylation of E-cadherin opens up possibilities to stop or revers the progression of scarring as a new strategy in trachoma treatment.
PB  - Institute of Biology and Immunology of Reproduction, BAS, COST Action CA16119 and Mouseprint Ltd.
C3  - Proceedings of CellFit meeting: In vitro 3-D total cell guidance and fitness; 2017 Sep 12-14; Albena Resort, Bulgaria
T1  - Chlamydia trachomatis infection triggers  Epithelial Mesenchymal Transition in  conjunctiva: involvement of DNA methylation
SP  - 64
EP  - 65
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5814
ER  - 

@conference{
author = "Rajić, Jovana and Grdović, Nevena and Inić-Kanada, Aleksandra and Stein, Elisabath and Schuerer, Nadine and Uskoković, Aleksandra and Ghasemian, Ehsan and Dinić, Svetlana and Mihailović, Mirjana and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Marija and Đorđević, Miloš and Poznanović, Goran and Barisani-Asenbauer, Talin and Vidaković, Melita",
year = "2017",
abstract = "Repeated eye infections with Chlamydia trachomatis (Ct) usually lead to conjunctiva! scarring, corneal damage and blindness (trachoma). Considering epithelial to mesenchymal transition (EMD as unavoidable player in the development of fibrosis and taking into account that EMT is epigenetically reg­ulated process, our intention was to explore whether Ct infec­tion induces EMT in vitro and to uncover potential underlying epigenetic mechanisms. 
Human conjunctiva! epithelial cells were infected with 107 IFU of Ct for 72 h. The mRNA and protein expression of EMT mark­ers (E-cadherin and fibronectin) were assessed by RT-qPCR, lmmunoblotting and lmmunocytochemfstry. DNA methyfation analysis of selected regions of marker genes were examined by MSP, HRM and Bs-Seq. 
The Ct infection induced EMT-related changes in mRNA and protein expression, downregulation of epithelial marker E-cadherin and upregulation of mesenchymal marker fibronectin. Increase in DNA methylation of E-cadherin gene promoter was in correlation with its decreased expression, while DNA meth­ylation status of fibronectin gene couldn't be related to its ex­pression level. 
We suggest that manipulating the EMT process via modula­tion of DNA methylation of E-cadherin opens up possibilities to stop or revers the progression of scarring as a new strategy in trachoma treatment.",
publisher = "Institute of Biology and Immunology of Reproduction, BAS, COST Action CA16119 and Mouseprint Ltd.",
journal = "Proceedings of CellFit meeting: In vitro 3-D total cell guidance and fitness; 2017 Sep 12-14; Albena Resort, Bulgaria",
title = "Chlamydia trachomatis infection triggers  Epithelial Mesenchymal Transition in  conjunctiva: involvement of DNA methylation",
pages = "64-65",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5814"
}

Rajić, J., Grdović, N., Inić-Kanada, A., Stein, E., Schuerer, N., Uskoković, A., Ghasemian, E., Dinić, S., Mihailović, M., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Đorđević, M., Poznanović, G., Barisani-Asenbauer, T.,& Vidaković, M.. (2017). Chlamydia trachomatis infection triggers  Epithelial Mesenchymal Transition in  conjunctiva: involvement of DNA methylation. in Proceedings of CellFit meeting: In vitro 3-D total cell guidance and fitness; 2017 Sep 12-14; Albena Resort, Bulgaria
Institute of Biology and Immunology of Reproduction, BAS, COST Action CA16119 and Mouseprint Ltd.., 64-65.
https://hdl.handle.net/21.15107/rcub_ibiss_5814

Rajić J, Grdović N, Inić-Kanada A, Stein E, Schuerer N, Uskoković A, Ghasemian E, Dinić S, Mihailović M, Arambašić Jovanović J, Tolić A, Đorđević M, Đorđević M, Poznanović G, Barisani-Asenbauer T, Vidaković M. Chlamydia trachomatis infection triggers  Epithelial Mesenchymal Transition in  conjunctiva: involvement of DNA methylation. in Proceedings of CellFit meeting: In vitro 3-D total cell guidance and fitness; 2017 Sep 12-14; Albena Resort, Bulgaria. 2017;:64-65.
https://hdl.handle.net/21.15107/rcub_ibiss_5814 .

Rajić, Jovana, Grdović, Nevena, Inić-Kanada, Aleksandra, Stein, Elisabath, Schuerer, Nadine, Uskoković, Aleksandra, Ghasemian, Ehsan, Dinić, Svetlana, Mihailović, Mirjana, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Marija, Đorđević, Miloš, Poznanović, Goran, Barisani-Asenbauer, Talin, Vidaković, Melita, "Chlamydia trachomatis infection triggers  Epithelial Mesenchymal Transition in  conjunctiva: involvement of DNA methylation" in Proceedings of CellFit meeting: In vitro 3-D total cell guidance and fitness; 2017 Sep 12-14; Albena Resort, Bulgaria (2017):64-65,
https://hdl.handle.net/21.15107/rcub_ibiss_5814 .

TY  - JOUR
AU  - Rajić, Jovana
AU  - Inic-Kanada, Aleksandra
AU  - Stein, Elisabeth
AU  - Dinić, Svetlana
AU  - Schuerer, Nadine
AU  - Uskoković, Aleksandra
AU  - Ghasemian, Ehsan
AU  - Mihailović, Mirjana
AU  - Vidaković, Melita
AU  - Grdović, Nevena
AU  - Barisani-Asenbauer, Talin
PY  - 2017
UR  - http://journal.frontiersin.org/article/10.3389/fcimb.2017.00253/full
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2836
AB  - Chlamydia trachomatis (Ct) can induce scarring disease of the ocular mucosa, known as trachoma, the most common infectious cause of blindness worldwide. We hypothesized that epithelial-mesenchymal transition (EMT) contributes to the fibrotic process in trachomatous scarring. Infection of human conjunctival epithelial cells (HCjE) with Ct activated signaling pathways involved in EMT induction, which was correlated with decreased expression of E-cadherin, guardian of the epithelial phenotype. In addition, Ct infection was associated with increased expression of two mesenchymal cell markers: fibronectin and α-SMA. The DNA methylation statuses of selected regions of E-cadherin, fibronectin, and α-SMA genes revealed that Ct infection was accompanied with changes in DNA methylation of the E-cadherin promoter, while the expression of the two mesenchymal markers was not related with this epigenetic event. Our data suggest that Ct infection of conjunctival epithelial cells induces EMT-like changes that go along with modification of the methylation profile of the E-cadherin promoter and could, as one of the earliest events, contribute to processes triggering conjunctival scarring.
T2  - Frontiers in Cellular and Infection Microbiology
T1  - Chlamydia trachomatis Infection Is Associated with E-Cadherin Promoter Methylation, Downregulation of E-Cadherin Expression, and Increased Expression of Fibronectin and α-SMA—Implications for Epithelial-Mesenchymal Transition
IS  - JUN
VL  - 7
DO  - 10.3389/fcimb.2017.00253
SP  - 253
EP  - 253
ER  - 

@article{
author = "Rajić, Jovana and Inic-Kanada, Aleksandra and Stein, Elisabeth and Dinić, Svetlana and Schuerer, Nadine and Uskoković, Aleksandra and Ghasemian, Ehsan and Mihailović, Mirjana and Vidaković, Melita and Grdović, Nevena and Barisani-Asenbauer, Talin",
year = "2017",
abstract = "Chlamydia trachomatis (Ct) can induce scarring disease of the ocular mucosa, known as trachoma, the most common infectious cause of blindness worldwide. We hypothesized that epithelial-mesenchymal transition (EMT) contributes to the fibrotic process in trachomatous scarring. Infection of human conjunctival epithelial cells (HCjE) with Ct activated signaling pathways involved in EMT induction, which was correlated with decreased expression of E-cadherin, guardian of the epithelial phenotype. In addition, Ct infection was associated with increased expression of two mesenchymal cell markers: fibronectin and α-SMA. The DNA methylation statuses of selected regions of E-cadherin, fibronectin, and α-SMA genes revealed that Ct infection was accompanied with changes in DNA methylation of the E-cadherin promoter, while the expression of the two mesenchymal markers was not related with this epigenetic event. Our data suggest that Ct infection of conjunctival epithelial cells induces EMT-like changes that go along with modification of the methylation profile of the E-cadherin promoter and could, as one of the earliest events, contribute to processes triggering conjunctival scarring.",
journal = "Frontiers in Cellular and Infection Microbiology",
title = "Chlamydia trachomatis Infection Is Associated with E-Cadherin Promoter Methylation, Downregulation of E-Cadherin Expression, and Increased Expression of Fibronectin and α-SMA—Implications for Epithelial-Mesenchymal Transition",
number = "JUN",
volume = "7",
doi = "10.3389/fcimb.2017.00253",
pages = "253-253"
}

Rajić, J., Inic-Kanada, A., Stein, E., Dinić, S., Schuerer, N., Uskoković, A., Ghasemian, E., Mihailović, M., Vidaković, M., Grdović, N.,& Barisani-Asenbauer, T.. (2017). Chlamydia trachomatis Infection Is Associated with E-Cadherin Promoter Methylation, Downregulation of E-Cadherin Expression, and Increased Expression of Fibronectin and α-SMA—Implications for Epithelial-Mesenchymal Transition. in Frontiers in Cellular and Infection Microbiology, 7(JUN), 253-253.
https://doi.org/10.3389/fcimb.2017.00253

Rajić J, Inic-Kanada A, Stein E, Dinić S, Schuerer N, Uskoković A, Ghasemian E, Mihailović M, Vidaković M, Grdović N, Barisani-Asenbauer T. Chlamydia trachomatis Infection Is Associated with E-Cadherin Promoter Methylation, Downregulation of E-Cadherin Expression, and Increased Expression of Fibronectin and α-SMA—Implications for Epithelial-Mesenchymal Transition. in Frontiers in Cellular and Infection Microbiology. 2017;7(JUN):253-253.
doi:10.3389/fcimb.2017.00253 .

Rajić, Jovana, Inic-Kanada, Aleksandra, Stein, Elisabeth, Dinić, Svetlana, Schuerer, Nadine, Uskoković, Aleksandra, Ghasemian, Ehsan, Mihailović, Mirjana, Vidaković, Melita, Grdović, Nevena, Barisani-Asenbauer, Talin, "Chlamydia trachomatis Infection Is Associated with E-Cadherin Promoter Methylation, Downregulation of E-Cadherin Expression, and Increased Expression of Fibronectin and α-SMA—Implications for Epithelial-Mesenchymal Transition" in Frontiers in Cellular and Infection Microbiology, 7, no. JUN (2017):253-253,
https://doi.org/10.3389/fcimb.2017.00253 . .

TY  - JOUR
AU  - Spasojević, Dragica
AU  - Zmejkoski, Danica
AU  - Glamočlija, Jasmina
AU  - Nikolić, Miloš
AU  - Soković, Marina
AU  - Milošević, Verica
AU  - Jarić, Ivana
AU  - Stojanović, Marijana
AU  - Marinković, Emilija
AU  - Barisani-Asenbauer, Talin
AU  - Prodanović, Radivoje
AU  - Jovanović, Miloš
AU  - Radotić, Ksenija
PY  - 2016
UR  - http://linkinghub.elsevier.com/retrieve/pii/S0924857916302679
UR  - https://www.scopus.com/record/display.uri?eid=2-s2.0-85000995852&origin=SingleRecordEmailAlert&dgcid=scalert_sc_search_email&txGid=5EA0B882AD39CF37A9F768C8B10917B5.wsnAw8kcdt7IPYLO0V48gA%3A1#
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2480
AB  - Nowadays bacterial resistance to known antibiotics is a serious health problem. In order to achieve more efficient treatment, lately there is an effort to find new substances, such as certain biomaterials, that are non-toxic to humans with antibiotic potential. Lignins and lignin-derived compounds have been proposed to be good candidates for use in medicine and health maintenance. In this study, the antibacterial activity of the lignin model polymer dehydrogenate polymer (DHP) in alginate hydrogel (Alg) was studied. The obtained results show that DHP−Alg has strong antimicrobial activity against several bacterial strains and biofilms and does not have a toxic effect on human epithelial cells. These results strongly suggest its application as a wound healing agent or as an adjunct substance for wound treatments.
T2  - International Journal of Antimicrobial Agents
T1  - Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment
IS  - 6
VL  - 48
DO  - 10.1016/j.ijantimicag.2016.08.014
SP  - 732
EP  - 735
ER  - 

@article{
author = "Spasojević, Dragica and Zmejkoski, Danica and Glamočlija, Jasmina and Nikolić, Miloš and Soković, Marina and Milošević, Verica and Jarić, Ivana and Stojanović, Marijana and Marinković, Emilija and Barisani-Asenbauer, Talin and Prodanović, Radivoje and Jovanović, Miloš and Radotić, Ksenija",
year = "2016",
abstract = "Nowadays bacterial resistance to known antibiotics is a serious health problem. In order to achieve more efficient treatment, lately there is an effort to find new substances, such as certain biomaterials, that are non-toxic to humans with antibiotic potential. Lignins and lignin-derived compounds have been proposed to be good candidates for use in medicine and health maintenance. In this study, the antibacterial activity of the lignin model polymer dehydrogenate polymer (DHP) in alginate hydrogel (Alg) was studied. The obtained results show that DHP−Alg has strong antimicrobial activity against several bacterial strains and biofilms and does not have a toxic effect on human epithelial cells. These results strongly suggest its application as a wound healing agent or as an adjunct substance for wound treatments.",
journal = "International Journal of Antimicrobial Agents",
title = "Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment",
number = "6",
volume = "48",
doi = "10.1016/j.ijantimicag.2016.08.014",
pages = "732-735"
}

Spasojević, D., Zmejkoski, D., Glamočlija, J., Nikolić, M., Soković, M., Milošević, V., Jarić, I., Stojanović, M., Marinković, E., Barisani-Asenbauer, T., Prodanović, R., Jovanović, M.,& Radotić, K.. (2016). Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment. in International Journal of Antimicrobial Agents, 48(6), 732-735.
https://doi.org/10.1016/j.ijantimicag.2016.08.014

Spasojević D, Zmejkoski D, Glamočlija J, Nikolić M, Soković M, Milošević V, Jarić I, Stojanović M, Marinković E, Barisani-Asenbauer T, Prodanović R, Jovanović M, Radotić K. Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment. in International Journal of Antimicrobial Agents. 2016;48(6):732-735.
doi:10.1016/j.ijantimicag.2016.08.014 .

Spasojević, Dragica, Zmejkoski, Danica, Glamočlija, Jasmina, Nikolić, Miloš, Soković, Marina, Milošević, Verica, Jarić, Ivana, Stojanović, Marijana, Marinković, Emilija, Barisani-Asenbauer, Talin, Prodanović, Radivoje, Jovanović, Miloš, Radotić, Ksenija, "Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment" in International Journal of Antimicrobial Agents, 48, no. 6 (2016):732-735,
https://doi.org/10.1016/j.ijantimicag.2016.08.014 . .