Abstract:
With the dramatic rise in the aging population, researching age-related macular degeneration (AMD), especially the severe form neovascular AMD (nAMD), has become more important than ever. In this study, we found that collagen type X was increased in retina-choroid tissue of mice with laser-induced choroidal neovascularization (CNV) based on immunohistofluorescence. RNA sequencing and bioinformatic analyses were performed to compare the retina-choroid tissue complex of the CNV mouse model to normal controls. Collagen type X alpha 1 chain (Col10a1) was among the most significantly upregulated genes, and the results were validated with an animal model at the mRNA and protein levels by quantitative real-time polymerase chain reaction (qPCR) and western blotting, respectively. COL10A1 was also upregulated in human retinal microvascular endothelial cells (HRMECs), human umbilical vein endothelial cells (HUVECs), RPE19 cells and RF/6A cells under hypoxic conditions. Next, in vitro and in vivo experiments were performed to study the effect of COL10A1 on neovascularization. siRNA knockdown of COL10A1 suppressed the proliferation and tube formation ability of HRMECs under hypoxic conditions. Snail family transcriptional repressor 1 (SNAIL1) and angiopoietin-2 (ANGPT2) were downregulated in COL10A1 knockdown HRMECs under hypoxic conditions and thus were potential downstream genes. Significant decreases in CNV leakage and CNV lesion area, as assessed by fundus fluorescein angiography (FFA) and immunofluorescence of choroidal flat mounts, respectively, were observed in a mouse model intravitreally injected with anti-collagen X monoclonal antibody (mAb) compared to the controls. In conclusion, COL10A1 promotes CNV formation and may represent a new candidate target for the treatment and diagnosis of nAMD and other neovascular diseases.
摘要:
随着人口老龄化的急剧上升,研究年龄相关性黄斑变性(AMD),尤其是严重的新生血管性AMD(nAMD),变得比以往任何时候都重要。在这项研究中,我们发现,在激光诱导的脉络膜新生血管(CNV)小鼠的视网膜脉络膜组织中,X型胶原增加.进行RNA测序和生物信息学分析以将CNV小鼠模型的视网膜-脉络膜组织复合物与正常对照进行比较。胶原蛋白X型α1链(Col10a1)是最显著上调的基因之一,并通过定量实时聚合酶链反应(qPCR)和蛋白质印迹在mRNA和蛋白质水平的动物模型上验证结果,分别。COL10A1在人视网膜微血管内皮细胞(HRMEC)中也上调,人脐静脉内皮细胞(HUVECs),缺氧条件下的RPE19细胞和RF/6A细胞。接下来,进行了体外和体内实验以研究COL10A1对新生血管形成的影响。在低氧条件下,COL10A1的siRNA敲低抑制了HRMECs的增殖和成管能力。在低氧条件下,蜗牛家族转录抑制因子1(SNAIL1)和血管生成素2(ANGPT2)在COL10A1敲低HRMEC中下调,因此是潜在的下游基因。CNV渗漏和CNV病变面积显著减少,通过荧光素眼底血管造影(FFA)和脉络膜扁平支架的免疫荧光评估,分别,与对照相比,在玻璃体内注射抗胶原X单克隆抗体(mAb)的小鼠模型中观察到。总之,COL10A1可促进CNV的形成,并可能成为治疗和诊断nAMD和其他新生血管性疾病的新候选靶点。