The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology, Chongqing Eye Institute, and Chongqing Branch (Municipality Division) of National Clinical Research Center for Ocular Diseases, Chongqing, 400016, China.
Beijing Ophthalmology & Visual Sciences Key Laboratory, Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing, 100730, China.
J Neuroinflammation. 2024 Nov 6;21(1):289. doi: 10.1186/s12974-024-03279-1.
Retinal neovascularization (RNV) disease is one of the leading causes of blindness, yet the molecular underpinnings of this condition are not well understood. To delve into the critical aspects of cell-mediated angiogenesis, we analyzed our previously published single-cell data. Our analysis revealed that retinal pigment epithelium (RPE) cells serve a crucial promotional function in angiogenesis. RPE cells were regulated by N6-methyladenosine (m6A). Next, we detected several critical m6A methylase in hypoxic ARPE-19 cells and in oxygen-induced retinopathy (OIR) mice, our results revealed a significant decrease in the level of methyltransferase like 3 (METTL3). METTL3 specific inhibitor STM2457 intravitreal injection or METTL3 conditional knockout mice both showed a significantly reduced neovascularization area of retina. Additionally, the angiogenesis-related abilities of human retinal endothelial cells (HRECs) were diminished after co-cultured with ARPE-19 treated with STM2457 or sh-METTL3 in vitro. Furthermore, through the integration of Methylated RNA immunoprecipitation (MeRIP) sequencing and RNA sequencing, we discovered that the metabolic enzyme quinolinate phosphoribosyltransferase (QPRT) was directly modified by METTL3 and recognized by the YTH N6-methyladenosine RNA binding protein C1 (YTHDC1). Moreover, after over-expressing QPRT, the angiogenic abilities of HRECs were improved through the phosphorylated phosphatidylinositol-3-kinase (p-PI3K)/ phosphorylated threonine kinase (p-AKT) pathway. Collectively, our study provided a novel therapeutic target for retinal angiogenesis.
视网膜新生血管(RNV)疾病是导致失明的主要原因之一,但这种疾病的分子基础尚未得到很好的理解。为了深入研究细胞介导的血管生成的关键方面,我们分析了之前发表的单细胞数据。我们的分析表明,视网膜色素上皮(RPE)细胞在血管生成中起关键的促进作用。RPE 细胞受到 N6-甲基腺苷(m6A)的调节。接下来,我们在缺氧的 ARPE-19 细胞和氧诱导的视网膜病变(OIR)小鼠中检测到几个关键的 m6A 甲基转移酶,我们的结果显示,甲基转移酶样蛋白 3(METTL3)的水平显著降低。METTL3 特异性抑制剂 STM2457 玻璃体内注射或 METTL3 条件性敲除小鼠均显示视网膜新生血管化面积显著减少。此外,与用 STM2457 或 sh-METTL3 处理的 ARPE-19 共培养后,人视网膜内皮细胞(HRECs)的血管生成相关能力降低。此外,通过整合甲基化 RNA 免疫沉淀(MeRIP)测序和 RNA 测序,我们发现代谢酶喹啉酸磷酸核糖基转移酶(QPRT)被 METTL3 直接修饰,并被 YTH N6-甲基腺苷 RNA 结合蛋白 C1(YTHDC1)识别。此外,过表达 QPRT 后,HRECs 的血管生成能力通过磷酸化磷脂酰肌醇-3-激酶(p-PI3K)/磷酸化丝氨酸/苏氨酸激酶(p-AKT)通路得到改善。总之,我们的研究为视网膜血管生成提供了一个新的治疗靶点。