VEGF receptor-1-mediated protection in dilated cardiomyopathy

VEGF 受体 1 介导的扩张型心肌病保护作用

• 批准号: 8257204
• 负责人: FABIO A RECCHIA
• 金额: $ 43.58万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-02 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8257204
• 关键词: 3' Untranslated Regions; Adenovirus Vector; Agonist; Angiogenic Factor; Angiotensin II; Apoptosis; Atrial Natriuretic Factor; Attenuated; Binding; CMV promoter; Candidate Disease Gene; Canis familiaris; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cells; Coronary Arteriosclerosis; Coronary Occlusions; Cytoprotection; Dilated Cardiomyopathy; Disease; Endothelial Cells; Etiology; Family; Family member; Fibrosis; Functional disorder; Future; Gene Delivery; Gene Transfer; Goals; Growth Factor; Heart; Heart Diseases; Human; In Vitro; Incubated; Innovative Therapy; Ligands; Malignant - descriptor; Mammals; Mediating; Mediator of activation protein; MicroRNAs; Modeling; Molecular; Muscle Cells; Myocardial; Names; Necrosis; Oxidative Stress; Pharmacological Treatment; Placental Growth Factor; Pre-Clinical Model; Protein Isoforms; Protocols documentation; Receptor Protein-Tyrosine Kinases; Regulatory Element; Role; Staging; Stimulation of Cell Proliferation; Stimulus; Testing; Time; Tissues; Transgenes; Transplantation; Untranslated Regions; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor B; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factors; Vascular Permeabilities; angiogenesis; base; cell type; clinical application; clinically relevant; coronary perfusion; gene therapy; in vivo; innovation; instrument; member; migration; novel therapeutics; promoter; receptor; research study; therapeutic transgene; transgene expression; vector

DESCRIPTION (provided by applicant): In mammals, the vascular endothelial growth factor (VEGF) family consists of five members, VEGF-A, -B, -C, - D and placenta growth factor (PlGF), synthesized by various cell types, including cardiomyocytes. VEGF-B is relatively understudied and yet is emerging as a major pro-survival factor. Similar to PlGF, it binds selectively the receptor VEGFR-1. We have recently found that VEGRF-1 is expressed in cardiomyocytes and provided the first evidence that VEGF-B167 gene transfer reduces myocyte apoptosis and angiotensin II-induced oxidative stress and markedly attenuates cardiac remodeling and functional derangement in dogs with pacing- induced dilated cardiomyopathy. On the other hand, VEGF-A gene delivery did not yield similar beneficial effects, in vivo. The cytoprotective/antiapoptotic and minimally angiogenic effects of VEGF167 render this factor an appealing candidate for gene therapy of non-ischemic dilated cardiomyopathy, which is not caused by coronary artery disease, hence would not benefit from angiogenesis. The overall goal of the present project is to test the hypothesis that VEGFR-1 is the main mediator of VEGF-induced cytoprotection in hearts with non-ischemic dilated cardiomyopathy. Cardiac gene transfer of VEGFR-1 ligands will be performed via adeno-associated vector-9 (AAV9) in chronically instrumented dogs with pacing-induced dilated cardiomyopathy. Parallel experiments will be conducted in vitro to explore molecular mechanisms and to optimize transgene constructs. Aim 1 is to determine the pathophysiological and cellular mechanisms underlying the different effects of VEGFR-1 versus VEGFR-2 stimulation in dilated cardiomyopathy. We will first identify the most cardioprotective VEGFR-1 ligand and then compare it to VEGF-E, a selective VEGFR-2 ligand, and to VEGF-A, a dual ligand, in paced hearts. The molecular basis of VEGFR-1 and VEGFR-2- mediated myocardial protection will be studied in cultured cardiomyocytes. Aim 2 is to determine the optimal timing for VEGF-B-induced cardioprotection during the course of pacing-induced dilated cardiomyopathy. The most cardioprotective VEGFR-1 ligand identified in specific aim 1 will be delivered as an AAV-carried transgene to the heart at different time points during the pacing protocol. Aim 3 is to test the cardioprotective effects of VEGF-B transgenes inducible by endogenous stimuli specifically occurring in the failing heart. An ideal strategy for future clinical applications would be to deliver therapeutic transgenes expressed only in the presence of molecular alterations relevant to the disease. Toward this goal, we will develop the following innovative strategies to achieve VEGF-B transgene expression induced by endogenous stimuli occurring in the failing heart: a) construction of artificial promoters comprising regulatory elements sensitive to oxidative stress; b) utilization of the natural promoter of the atrial natriuretic peptide; c) inclusion, in the 3' UTR region of the transgene, of target sequences for microRNAs that are expressed in the healthy heart, but downregulated in the failing heart. PUBLIC HEALTH RELEVANCE: This project will explore the role of growth factors, named vascular endothelial growth factors (VEGF), in the protection of cardiac cells under a pathological condition known as dilated cardiomyopathy. This cardiac disease is less frequent that others due to coronary occlusion or dysfunction, however it is very malignant, difficult to treat with the available pharmacological agents, and is responsible for more than 50% of cardiac transplants in the US. The administration of VEGFs can be a valid alternative to the current therapies for dilated cardiomyopathy, but, before this new strategy can be implemented, it will be necessary to understand in detail the function and beneficial of these factors as well as their harmful effects. We will therefore utilize a clinically relevant dog model of dilated cardiomyopathy and will test various members of the VEGF family that activate different receptors. Our hypothesis is that those VEGF members that activate the so called VEGF receptor-1 are the ones more likely to display relevant beneficial effects and to be considered for an innovative therapy of dilated cardiomyopathy.

描述（由申请人提供）：在哺乳动物中，血管内皮生长因子（VEGF）家族由五个成员组成，VEGF-A、-B、-C、-D和胎盘生长因子（PlGF），由各种细胞类型合成，包括心肌细胞。 VEGF-B 的研究相对较少，但正在成为主要的促生存因素。与 PlGF 类似，它选择性地结合受体 VEGFR-1。我们最近发现 VEGRF-1 在心肌细胞中表达，并提供了第一个证据，证明 VEGF-B167 基因转移可减少心肌细胞凋亡和血管紧张素 II 诱导的氧化应激，并显着减轻起搏引起的扩张型心肌病狗的心脏重塑和功能紊乱。另一方面，VEGF-A 基因递送在体内并未产生类似的有益效果。 VEGF167 的细胞保护/抗凋亡和最小的血管生成作用使该因子成为非缺血性扩张型心肌病基因治疗的有吸引力的候选者，这种心肌病不是由冠状动脉疾病引起的，因此不会从血管生成中受益。本项目的总体目标是检验以下假设：VEGFR-1 是 VEGF 诱导的非缺血性扩张型心肌病心脏细胞保护的主要介质。 VEGFR-1 配体的心脏基因转移将通过腺相关载体 9 (AAV9) 在患有起搏诱发的扩张型心肌病的长期仪器狗中进行。将在体外进行平行实验，以探索分子机制并优化转基因构建体。目标 1 是确定 VEGFR-1 与 VEGFR-2 刺激对扩张型心肌病产生不同影响的病理生理学和细胞机制。我们将首先确定最具心脏保护作用的 VEGFR-1 配体，然后将其与 VEGF-E（一种选择性 VEGFR-2 配体）和 VEGF-A（一种双配体）在起搏心脏中进行比较。将在培养的心肌细胞中研究 VEGFR-1 和 VEGFR-2 介导的心肌保护的分子基础。目标 2 是确定在起搏引起的扩张型心肌病过程中 VEGF-B 诱导的心脏保护作用的最佳时机。具体目标 1 中确定的最具心脏保护作用的 VEGFR-1 配体将作为 AAV 携带的转基因在起搏方案期间的不同时间点递送至心脏。目标 3 是测试由衰竭心脏中特异发生的内源性刺激诱导的 VEGF-B 转基因的心脏保护作用。未来临床应用的理想策略是提供仅在存在与疾病相关的分子改变时表达的治疗性转基因。为了实现这一目标，我们将开发以下创新策略来实现衰竭心脏中发生的内源刺激诱导的 VEGF-B 转基因表达： a) 构建包含对氧化应激敏感的调节元件的人工启动子； b) 心房钠尿肽天然启动子的利用； c) 在转基因的 3'UTR 区域中包含在健康心脏中表达但在衰竭心脏中下调的 microRNA 的靶序列。 公共健康相关性：该项目将探讨生长因子（血管内皮生长因子 (VEGF)）在扩张型心肌病病理条件下保护心肌细胞的作用。与其他因冠状动脉闭塞或功能障碍导致的心脏病相比，这种心脏病的发病率较低，但其恶性程度很高，很难用现有的药物治疗，并且在美国超过 50% 的心脏移植手术都是由这种心脏病引起的。 VEGF 的施用可以成为当前扩张型心肌病疗法的有效替代方案，但是，在实施这一新策略之前，有必要详细了解这些因素的功能和益处以及它们的有害影响。因此，我们将利用临床相关的扩张型心肌病狗模型，并测试激活不同受体的 VEGF 家族的各个成员。我们的假设是，那些激活所谓 VEGF 受体 1 的 VEGF 成员更有可能表现出相关的有益作用，并被考虑用于扩张型心肌病的创新疗法。