Knockout of AMD-associated gene POLDIP2 reduces mitochondrial superoxide in human retinal pigment epithelial cells

被引：0

作者：

Nguyen, Tu ^{[1
,2
]}

Urrutia-Cabrera, Daniel ^{[1
,2
]}

Wang, Luozixian ^{[1
,2
]}

Lees, Jarmon G. ^{[3
,4
,5
]}

Wang, Jiang-Hui ^{[1
,2
]}

Hung, Sandy S. C. ^{[1
,2
]}

Hewitt, Alex W. ^{[1
,2
,6
]}

Edwards, Thomas L. ^{[1
,2
]}

McLenachan, Sam ^{[7
]}

Chen, Fred K. ^{[1
,2
,7
]}

Lim, Shiang Y. ^{[3
,4
,5
]}

Luu, Chi D. ^{[1
,2
]}

Guymer, Robyn ^{[1
,2
]}

Wong, Raymond C. B. ^{[1
,2
]}

机构：

[1] Royal Victorian Eye & Ear Hosp, Ctr Eye Res Australia, Melbourne, Vic, Australia

[2] Univ Melbourne, Dept Surg, Ophthalmol, Melbourne, Vic, Australia

[3] St Vincents Inst Med Res, OBrien Inst Dept, Melbourne, Vic, Australia

[4] Univ Melbourne, Dept Surg, Melbourne, Vic, Australia

[5] Univ Melbourne, Dept Med, Melbourne, Vic, Australia

[6] Univ Tasmania, Menzies Inst Med Res, Sch Med, Hobart, Tas, Australia

[7] Univ Western Australia, Royal Perth Hosp, Dept Ophthalmol, Ctr Ophthalmol & Visual Sci,Incorporating Lions E, Perth, WA, Australia

来源：

AGING-US | 2023年 / 15卷 / 06期

基金：

英国医学研究理事会;

关键词：

age-related macular degeneration; retina; CRISPR/Cas; mitochondria superoxide; POLDIP2; DELTA-INTERACTING PROTEIN-2; COMPLEMENT FACTOR-H; MACULAR DEGENERATION; RPE; DRUSEN; ACTIVATION; DISEASE; ARPE-19; MODELS; RISK;

D O I：

暂无

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

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.

引用

页码：1713 / 1733

页数：21