Journal
AGING-US
Volume 15, Issue 6, Pages 1713-1733Publisher
IMPACT JOURNALS LLC
Keywords
age-related macular degeneration; retina; CRISPR/Cas; mitochondria superoxide; POLDIP2
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This study generated a POLDIP2 knockout cell line in human retinal pigment epithelium cells using CRISPR/Cas technology. It found a link between POLDIP2 and age-related macular degeneration (AMD), and showed that the knockout of POLDIP2 does not affect cell proliferation, cell viability, phagocytosis, and autophagy. RNA sequencing revealed significant changes in genes related to immune response, complement activation, oxidative damage, and vascular development in the POLDIP2 knockout cells. Additionally, the study demonstrated that loss of POLDIP2 reduces mitochondrial superoxide levels and upregulates mitochondrial superoxide dismutase SOD2 in ARPE-19 cells, supporting a potential role of POLDIP2 in regulating oxidative stress in AMD pathology.
Genetic and epidemiologic studies have significantly advanced our understanding of the genetic factors contributing to age-related macular degeneration (AMD). In particular, recent expression quantitative trait loci (eQTL) studies have highlighted POLDIP2 as a significant gene that confers risk of developing AMD. However, the role of POLDIP2 in retinal cells such as retinal pigment epithelium (RPE) and how it contributes to AMD pathology are unknown. Here we report the generation of a stable human RPE cell line ARPE-19 with POLDIP2 knockout using CRISPR/Cas, providing an in vitro model to investigate the functions of POLDIP2. We conducted functional studies on the POLDIP2 knockout cell line and showed that it retained normal levels of cell proliferation, cell viability, phagocytosis and autophagy. Also, we performed RNA sequencing to profile the transcriptome of POLDIP2 knockout cells. Our results highlighted significant changes in genes involved in immune response, complement activation, oxidative damage and vascular development. We showed that loss of POLDIP2 caused a reduction in mitochondrial superoxide levels, which is consistent with the upregulation of the mitochondrial superoxide dismutase SOD2. In conclusion, this study demonstrates a novel link between POLDIP2 and SOD2 in ARPE-19, which supports a potential role of POLDIP2 in regulating oxidative stress in AMD pathology.
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