Epithelial membrane protein 2 (EMP2) regulates hypoxia-induced angiogenesis in the adult retinal pigment epithelial cell lines.
Epithelial membrane protein 2 (EMP2) regulates hypoxia-induced angiogenesis in the adult retinal pigment epithelial cell lines.
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DOI：
10.1038/s41598-022-22696-x
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发表时间：
2022-11-12
影响因子：
4.6
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中科院分区：
文献类型：
--
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病理性视网膜新生血管形成是许多常见疾病（包括糖尿病视网膜病变、早产儿视网膜病变和视网膜血管阻塞性疾病）中可能导致失明的后果。本研究探讨上皮膜蛋白2（EMP2）及其在缺氧条件下作为人视网膜色素上皮（RPE）中血管生成的可能调节因子的作用。为了研究其影响，对RPE细胞系ARPE - 19进行基因改造，使其要么过表达EMP2，要么降低其水平，并分别进行RNA测序和蛋白质印迹分析以确认RNA和蛋白质水平的表达变化。在常氧条件和缺氧应激下评估蛋白质表达。用人脐静脉内皮细胞（HUVEC）进行毛细血管管形成试验以评估功能反应。发现EMP2表达在缺氧条件下在mRNA和蛋白质水平上均与促血管生成因子HIF1α和VEGF的表达呈正相关。从机制上讲，EMP2通过下调冯·希佩尔·林道蛋白（pVHL）来稳定HIF1α的表达。还发现EMP2介导的ARPE - 19细胞变化改变了条件培养基中一种或多种旁分泌因子的分泌，这些因子在体外功能性血管生成试验中可调节HUVEC迁移和毛细血管管形成。本研究确定EMP2是一种人RPE细胞系中血管生成的潜在介质。EMP2水平与促血管生成介质HIF1α和VEGF呈正相关，并且从机制上讲，EMP2通过下调pVHL调节HIF1α。本研究支持进一步研究EMP2作为视网膜病理性新生血管形成治疗的一个有希望的新靶点。
Pathologic retinal neovascularization is a potentially blinding consequence seen in many common diseases including diabetic retinopathy, retinopathy of prematurity, and retinal vaso-occlusive diseases. This study investigates epithelial membrane protein 2 (EMP2) and its role as a possible modulator of angiogenesis in human retinal pigment epithelium (RPE) under hypoxic conditions. To study its effects, the RPE cell line ARPE-19 was genetically modified to either overexpress EMP2 or knock down its levels, and RNA sequencing and western blot analysis was performed to confirm the changes in expression at the RNA and protein level, respectively. Protein expression was evaluated under both normoxic conditions or hypoxic stress. Capillary tube formation assays with human umbilical vein endothelial cells (HUVEC) were used to evaluate functional responses. EMP2 expression was found to positively correlate with expression of pro-angiogenic factors HIF1α and VEGF at both mRNA and protein levels under hypoxic conditions. Mechanistically, EMP2 stabilized HIF1α expression through downregulation of von Hippel Lindau protein (pVHL). EMP2 mediated changes in ARPE-19 cells were also found to alter the secretion of a paracrine factor(s) in conditioned media that can regulate HUVEC migration and capillary tube formation in in vitro functional angiogenesis assays. This study identifies EMP2 as a potential mediator of angiogenesis in a human RPE cell line. EMP2 levels positively correlate with pro-angiogenic mediators HIF1α and VEGF, and mechanistically, EMP2 regulates HIF1α through downregulation of pVHL. This study supports further investigation of EMP2 as a promising novel target for therapeutic treatment of pathologic neovascularization in the retina.
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影响因子：
2
作者：
Telander DG;Morales SA;Mareninov S;Forward K;Gordon LK
通讯作者：
Gordon LK
影响因子：
1.5
作者：
Qazi, Yureeda;Maddula, Surekha;Ambati, Balamurali K.
通讯作者：
Ambati, Balamurali K.
影响因子：
3.7
作者：
Fu M;Rao R;Sudhakar D;Hogue CP;Rutta Z;Morales S;Gordon LK;Braun J;Goodglick L;Wadehra M
通讯作者：
Wadehra M
影响因子：
5.8
作者：
Walter, Wencke;Sanchez-Cabo, Fatima;Ricote, Mercedes
通讯作者：
Ricote, Mercedes
影响因子：
3.6
作者：
Wadehra, M;Sulur, GG;Goodglick, L
通讯作者：
Goodglick, L