Non-canonical VEGF receptor signaling regulates retinal neovascularization

非经典 VEGF 受体信号传导调节视网膜新生血管形成

• 批准号: 8722755
• 负责人: Michael Edwin Boulton
• 金额: $ 21.14万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8722755
• 关键词: Non; canonical; VEGF; receptor; signaling

Neovascular diseases of the eye include retinopathy of prematurity, proliferative diabetic retinopathy, and the exudative or "wet" form of age-related macular degeneration (AMD). Together these diseases affect all age groups and are the leading causes of vision impairment in developed nations. The collective evidence suggests that the vascular endothelial growth factor (VEGF) family is critical for ocular angiogensis and this has become a major target for therapeutic intervention. Investigation of VEGF action has largely focused on receptor binding events at or near the plasma membrane and subsequent activation of classical signal transduction cascades. However, it is now apparent from our work and others that the signaling mediated by ligands binding to the VEGFRs (VEGFR1 and VEGFR2) is much more complex and involves intracellular trafficking of VEGFRs. Our data shows that targeted subcellular translocation of VEGFRs to adherens/tight junctions (AJs/TJs) results in the VEGFRs regulating vascular permeability or, if translocated to the nucleus VEGFRs can regulate transcription of pro- and anti-angiogenic regulators. Preliminary data also indicate that the specific ratio of VEGFR1:VEGFR2 at specific cell sites (such as AJs/TJs and the nucleus) is critical in determining vascular permeability and angiogenesis. Furthermore, endosomal sorting, ¿-secretase and SUMOylation appear to be key regulators of VEGFR trafficking. Based on these observations we propose the following hypothesis: VEGF-driven vascular permeability and neovascularization are highly dependent on the targeted subcellular translocation of specific VEGFRs. Pharmacological or genetic manipulation of components of the endosomal trafficking pathway, ¿-secretase complex and/or SUMOylation will reduce vascular permeability and inhibit aberrant retinal and choroidal neovascularization. We will test this hypothesis through the following aims. In Aim 1, we will a) determine the origin of nuclear VEGFRs by characterizing the routes of VEGFR internalization and trafficking, b) identify the nuclear targets of VEGFR1 and VEGFR2 and assess how these targets contribute to angiogenesis, c) assess how the ratio of nuclear VEGFR1:VEGFR2 dictates the angiogenic outcome and d) identify the mechanism by which ¿- secretase, presenilin and/or sumoylation regulate trafficking of VEGFRs. In Aim 2, we will a) determine if translocation of VEGFRs to AJs and TJs is via membrane diffusion or endosomal trafficking and b) assess how the ratio of VEGFR1 and VEGFR2 at AJs and TJs changes in response to pro- and antiangiogenic factors, the junctional binding partners (e.g. VE-cadherin, ¿- catenin, claudin-5) involved and how this affects permeability. In Aim 3, we will a) assess the changes in VEGFR subcellular localization and VEGFR1:VEGFR2 ratio in mice which have undergone pharmacological or genetic modulation of transmembrane proteases and/or SUMOylation and determine if this can prevent VEGF-induced retinal vascular permeability and/or retinal or choroidal angiogenesis in mice and b) evaluate the modulation of novel nuclear signaling pathways identified in sub Aim 1B in mouse models of angiogenesis. Understanding this novel non-canonical VEGF signalling pathway(s) will provide new information on angiogenesis and allow development of a sustainable treatment strategy for AMD and diabetic retinopathy.

眼部的新生血管性疾病包括早产儿视网膜病变、增殖性糖尿病视网膜病变和视网膜病变。 视网膜病变和渗出性或“湿性”形式的年龄相关性黄斑变性（AMD）。一起 这些疾病影响所有年龄组，是发达国家视力受损的主要原因。 nations.这些证据表明，血管内皮生长因子（VEGF）家族 对于眼部血管生成至关重要，这已成为治疗干预的主要目标。 VEGF作用的研究主要集中在血浆或血浆附近的受体结合事件 膜和随后的经典信号转导级联的激活。但据 现在从我们的工作和其他人的工作中显而易见的是，由结合到细胞表面的配体介导的信号传导， VEGFRs（VEGFR 1和VEGFR 2）要复杂得多，并且涉及细胞内转运， VEGFRs。我们的数据表明，VEGF靶向亚细胞易位到粘附细胞/紧密粘附细胞， 血管连接（AJs/TJs）导致VEGF调节血管通透性，或者如果易位到血管连接， 核VEGF可以调节促血管生成和抗血管生成调节物的转录。初步 数据还表明，在特定的细胞位点（如AJs/TJs）的VEGFR 1：VEGFR 2的特定比率 和细胞核）对于决定血管渗透性和血管生成至关重要。此外，委员会认为， 内体分选、<$-分泌酶和SUMO化似乎是VEGFR的关键调节因子 贩卖人口基于这些观察，我们提出以下假设：VEGF驱动 血管通透性和新血管形成高度依赖于靶向的 特异性VEGF的亚细胞易位。药理学或遗传操作 内体运输途径的组分，β-分泌酶复合物和/或 SUMO化将降低血管通透性并抑制异常的视网膜和脉络膜 新生血管形成我们将通过以下目标来检验这一假设。目标1： 将a）通过表征VEGFR的途径来确定核VEGFR的来源， B）鉴定VEGFR 1和VEGFR 2的核靶标，并评估 这些靶点如何促进血管生成，c）评估细胞核的比例如何， VEGFR 1：VEGFR 2决定血管生成结果，d）确定血管生成的机制。 分泌酶、早老素和/或类小泛素化调节VEGF的运输。在目标2中，我们将a） 确定VEGF向AJs和TJ的易位是通过膜扩散还是内体扩散 和B）评估AJs和TJ中VEGFR 1和VEGFR 2的比例如何变化， 对促血管生成因子和抗血管生成因子的反应，连接结合配偶体（例如VE-钙粘蛋白， catenin，claudin-5）参与，以及这如何影响渗透性。在目标3中，我们将a）评估 小鼠中VEGFR亚细胞定位和VEGFR 1：VEGFR 2比例的变化， 经历跨膜蛋白酶的药理学或遗传学调节和/或 SUMO化，并确定这是否可以防止VEGF诱导的视网膜血管通透性和/或 小鼠视网膜或脉络膜血管生成和B）评价新的核信号传导的调节 在小鼠血管生成模型中的子目标1B中鉴定的途径。理解这部小说 非典型VEGF信号通路将提供关于血管生成的新信息， 制定AMD和糖尿病视网膜病变的可持续治疗策略。

项目成果

Repurposing antimalarial aminoquinolines and related compounds for treatment of retinal neovascularization.

• DOI: 10.1371/journal.pone.0202436
• 发表时间: 2018
• 期刊: PloS one
• 影响因子: 3.7
• 作者: McAnally D;Siddiquee K;Gomaa A;Szabo A;Vasile S;Maloney PR;Divlianska DB;Peddibhotla S;Morfa CJ;Hershberger P;Falter R;Williamson R;Terry DB;Farjo R;Pinkerton AB;Qi X;Quigley J;Boulton ME;Grant MB;Smith LH
• 通讯作者: Smith LH

Development of an anti-angiogenic therapeutic model combining scAAV2-delivered siRNAs and noninvasive photoacoustic imaging of tumor vasculature development.

• DOI: 10.1016/j.canlet.2012.11.016
• 发表时间: 2013-05-10
• 期刊: CANCER LETTERS
• 影响因子: 9.7
• 作者: Ruan, Qing;Xi, Lei;Boye, Sanford L.;Han, Song;Chen, Zhi J.;Hauswirth, William W.;Lewin, Alfed S.;Boulton, Michael E.;Law, Brian K.;Jiang, Wen G.;Jiang, Huabei;Cai, Jun
• 通讯作者: Cai, Jun

Consequences of oxidative stress in age-related macular degeneration.

• DOI: 10.1016/j.mam.2012.03.009
• 发表时间: 2012-08
• 期刊: MOLECULAR ASPECTS OF MEDICINE
• 影响因子: 10.6
• 作者: Jarrett, Stuart G.;Boulton, Michael E.
• 通讯作者: Boulton, Michael E.