Ramshekar Aniket, Asare-Bediako Bright, Nguyen Jasmine, Suresh Arundhathy, Simmons Aaron, Bretz Colin A, Wang Haibo, Kunz Eric, Zaugg Chandler J, Wallace-Carrete Chris, Hartnett M Elizabeth
Department of Ophthalmology, Byers Eye Institute Stanford University School of Medicine Stanford California USA.
John A. Moran Eye Center University of Utah Salt Lake City Utah USA.
FASEB Bioadv. 2025 Sep 10;7(9):e70051. doi: 10.1096/fba.2025-00146. eCollection 2025 Sep.
Vascular endothelial growth factor (VEGF) is important in both developmental and pathologic angiogenesis in retinopathy of prematurity (ROP). Using a rat model representative of ROP, we found that regulation of VEGF signaling through VEGF receptor 2 (VEGFR2) in retinal microvascular endothelial cells (RMVECs) extended developmental angiogenesis but reduced pathologic angiogenesis, that is, intravitreal neovascularization (IVNV). We identified an adaptor protein, MEMO1, in IVNV in the rat model and tested the hypothesis that MEMO1 in RMVECs was important in IVNV by regulating signaling through VEGFR2. Instead, we found MEMO1 knockdown enhanced phosphorylation of VEGF-induced VEGFR2 and STAT3 and increased wound closure in vitro using cultured human RMVECs. Furthermore, MEMO1 overexpression suppressed VEGF-induced VEGFR2 and STAT3 phosphorylation and dampened VEGF-induced RMVEC wound closure. In contrast, in the absence of VEGF, MEMO1 overexpression promoted RMVEC proliferation in the wound closure assay and AKT phosphorylation, supporting a role for MEMO1 in VEGF-independent angiogenic processes. In vivo, retinal endothelial cell-specific knockdown of MEMO1 in the rat ROP model significantly increased IVNV but did not affect developmental angiogenesis. Our findings support a novel regulatory role for MEMO1 where MEMO1 limits VEGF-driven IVNV and promotes VEGF-independent angiogenic signaling. These results suggest MEMO1 may serve as a protective modulator of pathological angiogenesis in ROP and represent a potential therapeutic target to limit IVNV while preserving physiologic angiogenesis.
血管内皮生长因子(VEGF)在早产儿视网膜病变(ROP)的发育性血管生成和病理性血管生成中均起重要作用。利用一种代表ROP的大鼠模型,我们发现,通过视网膜微血管内皮细胞(RMVECs)中的血管内皮生长因子受体2(VEGFR2)对VEGF信号进行调控,可延长发育性血管生成,但减少病理性血管生成,即玻璃体内新生血管形成(IVNV)。我们在大鼠模型的IVNV中鉴定出一种衔接蛋白MEMO1,并通过调控VEGFR2信号来测试RMVECs中的MEMO1在IVNV中起重要作用这一假设。相反,我们发现,使用培养的人RMVECs,敲低MEMO1可增强VEGF诱导的VEGFR2和STAT3磷酸化,并增加体外伤口愈合。此外,过表达MEMO1可抑制VEGF诱导的VEGFR2和STAT3磷酸化,并减弱VEGF诱导的RMVEC伤口愈合。相比之下,在无VEGF的情况下,过表达MEMO1可在伤口愈合试验中促进RMVEC增殖和AKT磷酸化,支持MEMO1在不依赖VEGF的血管生成过程中的作用。在体内,在大鼠ROP模型中对视网膜内皮细胞进行特异性敲低MEMO1可显著增加IVNV,但不影响发育性血管生成。我们的研究结果支持MEMO1具有一种新的调节作用,即MEMO1可限制VEGF驱动的IVNV,并促进不依赖VEGF的血管生成信号。这些结果表明,MEMO1可能是ROP中病理性血管生成的一种保护性调节因子,并且是在保留生理性血管生成的同时限制IVNV的潜在治疗靶点。