Mechanistic study and therapeutic application of AIBP in AMD

项目摘要 脉络膜新生血管（CNV），湿性型年龄相关性黄斑变性（AMD），是一种主要的 老年人失明的原因目前的抗血管内皮生长因子（VEGF）疗法具有许多优点。 严重的局限性包括：1）许多患者对单一抗VEGF治疗反应不佳或根本没有反应 （无应答者）; 2）即使对于应答者，长期结局也是次优的，并且可能导致抗VEGF抗体， 耐药性; 3）重复玻璃体内注射对生活质量产生不利影响，并增加局部 并发症因此，改善CNV治疗功效的替代疗法或联合疗法代表了一种新的治疗方法。 未满足的主要临床需求。CNV患者中抗VEGF抵抗通常与小动脉CNV相关 （大口径分支小动脉，血管环和吻合连接），其中巨噬细胞 已知发挥重要作用，而抗VEGF应答者的特征在于毛细血管CNV，其中 VEGFR 2信号传导具有压倒性的作用。分泌型载脂蛋白A-I结合蛋白（AIBP） 与其伴侣apoA-I或高密度脂蛋白（HDL）结合，以增强胆固醇流出并抑制脂筏- 在过度活跃的内皮细胞中锚定VEGFR 2信号传导。通过结合Toll样受体4（TLR 4），AIBP 增加巨噬细胞的胆固醇流出，从而抑制炎症（图1）。VEGF是一种 将巨噬细胞募集到分泌增加的炎症血管部位的炎性细胞因子 由巨噬细胞产生的额外的VEGF和其他促血管生成因子产生正反馈回路。我们 假设AIBP、apoA-I和抗VEGF组合将通过以下方式对抗抗VEGF抗性： 同时靶向VEGF、内皮细胞和巨噬细胞。我们最近发现， 治疗通过有效抑制小动脉CNV有效治疗抗VEGF抵抗，而阿柏西普 单药治疗无效（IOVS，2022）。我们的目标是开发AIBP/apoA-I/抗VEGF组合 本发明提供了一种应用广泛且长期有效的用于AMD的基因治疗，从而克服了现有基因治疗的局限性， Aim 1将决定AIBP在病理性血管生成中的作用。目标2将检验这样一个假设， 在CNV中，AIBP与巨噬细胞的联合治疗对于通过联合治疗克服抗VEGF抗性是必要的。目的 3将开发长期AIBP/apoA-I/抗VEGF联合基因治疗小鼠AMD模型中的CNV。

PROJECT SUMMARY Choroidal neovascularization (CNV), the wet type of age-related macular degeneration (AMD), is a major cause of blindness in the elderly. Current anti-vascular endothelial growth factor (VEGF) therapy has a number of serious limitations including: 1) many patients respond poorly or not at all to mono anti-VEGF treatments (non-responders); 2) the long-term outcomes are suboptimal even for responders and can lead to anti-VEGF resistance; 3) repetitive intravitreal injections adversely affect quality of life and increase the risk of local complications. Therefore, alternative or combination therapies to improve CNV treatment efficacy represent a major unmet clinical need. Anti-VEGF resistance in CNV patients is frequently associated with arteriolar CNV (large-caliber branching arterioles, vascular loops and anastomotic connections), in which macrophages are known to play important roles, whereas anti-VEGF responders are characterized by capillary CNV, in which VEGFR2 signaling has an overriding role. The secretory apolipoprotein A-I (apoA-I) binding protein (AIBP) binds to its partner apoA-I or high-density lipoprotein (HDL) to enhance cholesterol efflux and inhibit lipid raft- anchored VEGFR2 signaling in hyperactive endothelial cells. By binding the toll-like receptor 4 (TLR4), AIBP augments cholesterol efflux from macrophages, thereby suppressing inflammation (Fig. 1). VEGF is an inflammatory cytokine that recruits macrophages to vascular sites of inflammation where heightened secretion of additional VEGF and other pro-angiogenic factors by macrophages creates positive feedback loops. We hypothesize that combinations of AIBP, apoA-I and anti-VEGF will combat anti-VEGF resistance by simultaneously targeting VEGF, endothelial cells, and macrophages. We recently showed that this combination therapy was effective in treating anti-VEGF resistance by potently inhibiting arteriolar CNV, whereas aflibercept monotherapy was ineffective (IOVS, 2022). Our objective is to develop AIBP/apoA-I/anti-VEGF combination gene therapy with broad application and long-term efficacy for AMD, thereby obviating the limitations of current Aim1 will determine the role of AIBP in pathological angiogenesis. Aim 2 will test the hypothesis that targeting macrophages with AIBP is necessary to overcome anti-VEGF resistance in CNV by combination therapy. Aim 3 will develop long-term AIBP/apoA-I/anti-VEGF combination gene therapy for CNV in mouse AMD models.