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  - 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  - 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 . .