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 .