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Transforming growth factor β-related genes in human retinal pigment epithelial cells after tacrolimus treatment
Malgorzata Kimsa1, Barbara Strzalka-Mrozik2, Magdalena Kimsa-Dudek3
1Department of Biochemistry, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland.
Background:
The transforming growth factor β (TGFβ) family plays an important role in the pathogenesis of many diseases, including fibrotic pathologies of the eyes. The difficulties of surgical procedures contribute to the search for new treatment strategies for proliferative vitreoretinopathy. Therefore, the aim of this study was to investigate the expression profile of TGFβ isoforms, their receptors, and TGFβ-related genes in human retinal pigment epithelial cells (RPE) after tacrolimus (FK-506) treatment in the presence or absence of lipopolysaccharide (LPS)-induced inflammation.
Methods:
The expression profile was analyzed using oligonucleotide microarrays and quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) techniques.
Results:
Analysis using oligonucleotide microarrays revealed 20 statistically significant differentially expressed TGFβ-related genes after LPS treatment in relation to control cells, and after tacrolimus and LPS treatment in relation to LPS-treated cells. Moreover, our results showed that mRNA levels for TGFβ2 and TGFβR3 after tacrolimus treatment, and for TGFβR3 after tacrolimus and LPS treatment in RPE cells were decreased. In turn, in the presence of LPS-induced inflammation, TGFβ2 mRNA level was increased.
Conclusions:
These results can be important in regard to the treatment of proliferative vitreoretinopathy, pathogenesis of which is associated with processes regulated by TGFβ, such as inflammation, proliferation, epithelial-mesenchymal transition (EMT), and fibrosis.
Insights
Tacrolimus (FK-506) affects transforming growth factor β (TGFβ) gene expression in retinal cells, potentially impacting proliferative vitreoretinopathy treatment. It decreased TGFβ2 and TGFβR3 mRNA, but inflammation increased TGFβ2.
Area of Science:
- Ophthalmology
- Molecular Biology
- Immunology
Background:
- Transforming growth factor β (TGFβ) family is implicated in fibrotic eye diseases.
- Proliferative vitreoretinopathy (PVP) pathogenesis necessitates novel therapeutic strategies.
- Retinal pigment epithelial (RPE) cells are crucial in ocular homeostasis and disease.
Purpose of the Study:
- Investigate TGFβ isoform, receptor, and related gene expression in RPE cells.
- Evaluate the impact of tacrolimus (FK-506) on RPE cells.
- Examine tacrolimus effects under lipopolysaccharide (LPS)-induced inflammation.
Main Methods:
- Oligonucleotide microarrays for gene expression profiling.
- Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) for validation.
- Cultured human RPE cells treated with tacrolimus and/or LPS.
Main Results:
- 20 differentially expressed TGFβ-related genes identified post-LPS and post-tacrolimus/LPS treatment.
- Tacrolimus decreased TGFβ2 and TGFβR3 mRNA levels in RPE cells.
- LPS-induced inflammation increased TGFβ2 mRNA levels, while tacrolimus counteracted this for TGFβR3.
Conclusions:
- Findings offer insights into proliferative vitreoretinopathy treatment.
- TGFβ signaling pathways are key targets in managing inflammation, proliferation, EMT, and fibrosis in the eye.
- Tacrolimus may modulate TGFβ-driven processes relevant to fibrotic eye conditions.
