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，分别由VEGF -A，-B，-C，-C， - D和胎盘生长因子（PLGF）组成，由包括心脏肌细胞在内的各种细胞类型合成。 VEGF-B相对研究，但正在成为主要的生存因素。与PLGF相似，它选择性地结合了受体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基因转移减少了肌细胞凋亡和血管紧张素II诱导的氧化应激，并显着减轻了心脏重塑和功能性危害，并在狗中发挥了造成诱发的诱导诱导的诱导降低的心脏造成心脏体现的疾病。另一方面，VEGF-A基因递送在体内没有产生类似的有益作用。 VEGF167的细胞保护/抗凋亡和微小的血管生成作用使该因素成为非缺血性扩张性心肌病基因治疗的吸引力候选，这不是由冠状动脉疾病引起的，因此不会受益于血管生成。本项目的总体目的是检验以下假设：VEGFR-1是VEGF诱导的非缺血性扩张性心肌病的心脏中VEGF诱导的细胞保护作用的主要介体。 VEGFR-1配体的心脏基因转移将通过与起搏诱导的扩张性心肌病的长期仪器狗中的腺相关矢量9（AAV9）进行。平行实验将在体外进行，以探索分子机制并优化转基因构建体。目的1是确定在扩张心肌病中VEGFR-1与VEGFR-2刺激的不同作用背后的病理生理和细胞机制。我们将首先识别最具心脏保护性的VEGFR-1配体，然后将其与选择性的VEGF-2配体VEGF-E进行比较，并在节奏的心脏中与双重配体VEGF-A进行比较。 VEGFR-1和VEGFR-2介导的心肌保护的分子基础将在培养的心肌细胞中进行研究。 AIM 2是确定在起搏引起的扩张性心肌病过程中VEGF-B诱导的心脏保护的最佳时机。特定目标1中确定的最心脏保护性VEGFR-1配体将在起搏协议期间的不同时间点作为AAV婚姻转基因传递。 AIM 3是测试由特异性心脏中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与该疾病相关的分子改变的情况下表达的治疗转基因。为了实现这一目标，我们将开发以下创新策略，以实现失败心脏中发生的内源性刺激引起的VEGF-B转基因表达：a）构建人造启动子，包括对氧化应激敏感的调节元件； b）利用心房纳特里肽的天然启动子； c）包含在转基因的3'UTR区域中，在健康心脏中表达的microRNA的靶序列，但在失败的心脏中被下调。 公共卫生相关性：该项目将探索生长因子的作用，即称为血管内皮生长因子（VEGF），在病理疾病中保护心脏细胞的保护性心肌病变。由于冠状动脉阻塞或功能障碍，这种心脏疾病的频率较小，但是它非常恶性，很难用可用的药理剂治疗，并且负责美国50％以上的心脏移植。 VEGF的给药可以是当前扩张心肌病的疗法的有效替代方法，但是，在实施这种新策略之前，有必要详细了解这些因素的功能和有益，以及它们的有害影响。因此，我们将利用临床上相关的扩张心肌病模型，并将测试激活不同受体的VEGF家族的各种成员。我们的假设是，那些激活所谓的VEGF受体1的VEGF成员更有可能表现出相关的有益作用，并被认为是对扩张性心肌病的创新疗法。