Angiogenesis in Diseased Model

描述(由申请人提供)：糖尿病是心力衰竭(HF)患者最重要的并存疾病，它对心力衰竭和缺血性心脏病患者的预后产生不利影响。它的特点是侧支血管形成减少，以应对冠状动脉缺血事件。因此，在糖尿病的26周中，毛细血管密度和直径显示出超过20%的进行性下降。血管内皮生长因子(VEGF)是生理和病理条件下新血管形成的主要介质，在发育血管形成中起着至关重要的作用，在糖尿病并发症中被发现受到抑制。临床和实验性糖尿病均有冠状动脉侧支循环异常的报道。我们最初和继续研究缺血再灌流心脏氧化还原信号的动机使我们进入了糖尿病氧化还原调节和应激信号的新研究领域。硫氧还蛋白(Trx)是我们实验室最近发现的一种氧化还原活性蛋白，可诱导HO-1和血管内皮生长因子，从而保护心肌免受氧化应激的影响。Trx具有血管生成潜能，可能通过血管生成或缺血/梗死心肌血管生成促进新生血管的形成。我们这个项目的长期目标是了解糖尿病心力衰竭(HF)中氧化应激诱导的异常，并建立有效的治疗策略来治疗未来的糖尿病患者。因此，这项研究试图通过确定糖尿病动物中TRX-VEGF信号的潜在候选者来解决一个重要的临床问题。这项研究将利用广泛的多学科方法，结合各种技术、现代分子生物学、成像技术、基因治疗、基因靶向和生理学。目的I.糖尿病病情加重对氧化应激/氧化还原状态的影响将与小窝蛋白、Trx、HO-1和VEGF的表达及血管生成一起研究。目的II.硫氧还蛋白-1的过度表达包括HO-1、VEGF/VEGFR2和eNOS的表达增加，伴随着新生血管的增加。目的III.探讨GSK-3β和β-连环蛋白介导的血管内皮生长因子信号转导通路在糖尿病动物血管生成中的作用。目的IV.联合应用Ad-VEGF和Ad-Ang-1基因治疗可触发生存信号和新生血管形成。精心设计的链脲佐菌素诱导的糖尿病大鼠和基因工程小鼠模型的一系列实验将被使用。总之，这项拟议的研究将有助于我们理解与糖尿病相关疾病相关的血管生成调控机制，并可能为治疗缺血性疾病提供新的治疗策略。

DESCRIPTION (provided by applicant): Diabetes is the most significant co-morbidity of patients with heart failure (HF), which adversely affects outcomes in patients with HF and ischemic heart disease. It is characterized by a decreased collateral vessel formation in response to coronary ischemic events. Therefore capillary density and diameter exhibited progressive decreases of more than 20% over 26 week of diabetes. Vascular endothelial growth factor (VEGF) is a major mediator of neovascularization in physiological and pathological conditions with crucial roles in developmental blood vessel formation was found to be inhibited in diabetic complications. Abnormalities of the coronary collateral circulation have been reported in clinical and experimental diabetes mellitus. Our initial and continuing motivation to study of REDOX SIGNALING IN ISCHEMIC REPERFUSED heart leads us to step into a new research field of redox regulation and stress signal in diabetes mellitus. Thioredoxin (Trx) a redox active protein has recently been shown by our laboratory to induce HO-1 and VEGF that protects myocardium from oxidative stress. Trx has angiogenic potential, which may contribute to de novo development of vessels by vasculogenesis or angiogenesis in ischemic/infarcted myocardium. Our long-term goal of this project is to understand the oxidative stress induced abnormalities in diabetic heart failure (HF) and to establish effective therapeutic strategies to treat diabetic patients in future. Thus, this study will attempt to address an important clinical issue by identifying potential candidates of Trx-VEGF signaling in diabetic animals. This study will utilize a broad multidisciplinary approach that will combine various techniques, modern molecular biology, imaging technology, gene therapy, gene targeting and physiology. Aim I. The impact of oxidative stress/redox status with increased diabetic condition will be studied along with caveolin, Trx, HO-1 and VEGF expression and angiogenesis. Aim II. Overexpression of Thiredoxin-1 involves increased HO-1, VEGF/VEGFR2 and eNOS expression followed by increased neovascularization. Aim III. Involvement of GSK-3 beta and beta-catenin mediated VEGF signaling in diabetic animals will be explored related to angiogenesis. Aim IV. Combination gene therapy with Ad-VEGF and Ad-Ang- 1 triggers survival signal and neovascularization. Thoughtfully designed series of experiments in streptozotocin-induced diabetic rat and genetically engineered mouse model will be used. Collectively, the proposed study will contribute to our understanding of the mechanisms that regulate angiogenesis in association with diabetic-related disorders, and may provide novel therapeutic treatment strategies to cure ischemic disease.