OVEREXPRESSION OF ENOS IN CRITICAL LIMB ISCHEMIA

ENOS 在严重肢体缺血中的过度表达

• 批准号: 7179321
• 负责人: Louis Michael Messina
• 金额: $ 38.52万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7179321
• 关键词: Amputation; Angiogenic Proteins; Animal Model; Arterial Occlusive Diseases; Arteries; Blood Vessels; Blood flow; Catheters; Cells; Clinical; Condition; Dermal; Development; Diabetes Mellitus; Disease; Edema; Effectiveness; Elderly; Gangrene; Gene Delivery; Gene Transfer; Gene Transfer Techniques; Goals; Hindlimb; Human; Image; Impairment; Inflammatory; Ischemia; Lasers; Limb structure; Localized; Lower Extremity; Measures; Methods; Modeling; Molecular; Operative Surgical Procedures; Pain; Patients; Perfusion; Peripheral; Protein Overexpression; Rate; Rattus; Research; Research Proposals; Rest; Role; Secondary to; Site; Skeletal Muscle; Skin; Stem cells; Sustainable Development; Symptoms; System; Techniques; Testing; Therapeutic; Tissues; Ulcer; United States; Vascular Diseases; Vascular Endothelial Growth Factors; base; cigarette smoking; clinically relevant; concept; disability; gene therapy; human NOS3 protein; hypercholesterolemia; insight; interest; new growth; novel; novel therapeutics; success; tetrahydrobiopterin; transgene expression

DESCRIPTION (provided by applicant): Despite advances in the surgical and catheter-based treatments for critical limb ischemia (CLI), 150,000 patients require lower limb amputation in the United States annually. Believing that an effective molecular therapy would have a significant clinical impact on the disability and limb loss rates in these patients, this study looks specifically at the role of endothelial nitric oxide synthase (eNOS) in collateral artery development (arteriogenesis). Because such development can reverse CLI, our central hypothesis maintains that the overexpression of eNOS will induce the ordered and sustainable development of collateral arteries in a way that will reverse CLI. Specifically, this research proposal aims 1) to identify the optimal gene transfer condition in a rat hind limb mode of ischemia; 2) to examine whether these eNOS gene transfer techniques can also reverse CLI clinically relevant animal models of diabetes, advanced age and hypercholesterolemia, and cigarette smoking; and 3) to characterize the molecular mechanisms by which eNOS overexpression increases arteriogenesis. To accomplish these aims, our research plan employs novel gene delivery techniques and sites of delivery. In particular, a unique method of intra-arterial gene transfer under vascular isolation that results in highly efficient and localized transgene expression will be used for gene transfer and laser Doppler perfusion imaging to measure dermal blood flow will facilitate the accomplishment of the above aims. Understanding the mechanisms by which eNOS exerts its powerful effects on arteriogenesis should provide additional insight into the fundamental mechanisms regulating arteriogenesis, and therefore open new therapeutic approaches for the treatment of CLI and other vascular diseases.

描述（由申请人提供）： 尽管严重肢体缺血 (CLI) 的手术和基于导管的治疗取得了进展，但美国每年仍有 150,000 名患者需要下肢截肢。本研究相信有效的分子疗法将对这些患者的残疾和肢体丧失率产生重大的临床影响，因此专门研究了内皮一氧化氮合酶 (eNOS) 在侧支动脉发育（动脉生成）中的作用。因为这种发展可以逆转 CLI，我们的中心假设认为，eNOS 的过度表达将诱导侧支动脉有序且可持续的发展，从而逆转 CLI。具体来说，本研究计划的目的是：1）确定大鼠后肢缺血模式下的最佳基因转移条件； 2) 检查这些 eNOS 基因转移技术是否也可以逆转糖尿病、高龄和高胆固醇血症以及吸烟的 CLI 临床相关动物模型； 3) 表征 eNOS 过度表达促进动脉生成的分子机制。为了实现这些目标，我们的研究计划采用了新颖的基因传递技术和传递位点。特别是，在血管隔离下进行动脉内基因转移的独特方法可实现高效和局部转基因表达，将用于基因转移和激光多普勒灌注成像来测量真皮血流量将有助于实现上述目标。了解 eNOS 对动脉生成发挥强大作用的机制应该可以为调节动脉生成的基本机制提供更多的见解，从而为 CLI 和其他血管疾病的治疗开辟新的治疗方法。