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Sze Yuan Ho, Beiying Qiu, Gemmy Chui Ming Cheung, Tien Yin Wong, Wan Jin Hong, Xiaomeng Wang; The role of a novel metabolic regulator in human retinal pigment epithelial cell function. Invest. Ophthalmol. Vis. Sci. 2019;60(9):1947.
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Currently, anti-vascular endothelial growth factor (VEGF) therapy is the first line treatment for neovascular age-related macular degeneration (nAMD). However, a substantial number of patients are not responsive to the treatment or may develop resistance over time. Furthermore, subretinal fibrosis can develop in approximately 50% of patients after two years of anti-VEGF treatment. Epithelial-mesenchymal transition (EMT) in retinal pigment epithelial cells (RPECs) was reported to play a key role in subretinal fibrosis associated with nAMD. In this study, we aim to investigate the impact of a novel lipase, SIPRAD007, on RPEC function.
Trypan blue exclusion and Ki-67 proliferation assays were performed to study the role of SIPRAD007 on the viability and proliferation of immortalised human RPECs, ARPE-19. The effect of SIPRAD007 on TGFβ1-induced expression of cytostatic and fibrotic genes was studied by real-time quantitative polymerase chain reaction (RT-qPCR) and/or Western blot analysis.
Our study showed a significant reduction in the viability and proliferation of SIPRAD007 siRNA transfected ARPE-19 cells as compared to scrambled siRNA-treated control cells by 14.2 ± 2.7% (n=4) and 46.3 ± 12.9% (n=3) respectively. This observation was supported by an increased expression of cell cycle inhibitors, p15 and p21, in SIPRAD007 knockdown ARPE-19 cells. Furthermore, the siSIPRAD007-treated ARPE-19 cells were more responsive to TGFβ1-induced expression of connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1).
Our results demonstrated an important role of SIPRAD007 in RPEC viability, proliferation and the expression of fibrotic markers, including PAI-1 and CTGF.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
The protein levels of CTGF and PAI-1 were significantly upregulated in TGFβ1-treated siSIPRAD007 transfected ARPE-19 cells. Ordinary one-way ANOVA followed by Tukey’s multiple comparisons test was used for statistical analysis. *P < 0.05. Data are expressed as mean ± SEM (n ≥ 3).
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