iNOS gene therapy to prevent allograft vasculopathy

iNOS 基因治疗预防同种异体移植血管病变

• 批准号: 6668348
• 负责人: RICHARD L. SIMMONS
• 金额: $ 29.24万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6668348
• 关键词: apoptosis; atherosclerosis; biotechnology; clinical research; cooperative study; gene therapy; heart transplantation; human subject; isozymes; laboratory rat; miniature swine; nitric oxide synthase; transfection /expression vector; transplant rejection

Based on current data from the United Network for Organ Sharing (UNOS), cardiac allograft vasculopathy (CAV) is the leading cause of death after the first year of transplantation and accounts for 25% of all deaths annually after the third year. Initial studies from our laboratory have demonstrated that the expression of inducible nitric oxide synthase (iNOS; NOS2) can suppress the development of neointimal hyperplasia and prevent the development of CAV. The proposal described herein will test the hypothesis that viral vectors capable of transferring the human iNOS gene can suppress the development of CAV without causing undue toxicity to cardiac myocytes. Furthermore, this proposal will serve as the pre-clinical basis for near-term clinical trials for the future prevention of CAV in humans. In order to serve these ends, we will carry out the following Specific Aims: 1) to determine the efficacy of several viral vectors (adenoviral, adeno-associated virus, and lentivirus) containing the human iNOS gene (Ad-iNOS, AAV-iNOS, Lt-iNOS) and their respective control vectors for suppression of cardiac transplant arteriosclerosis in the rat model; 2) to determine toxicity of Ad-iNOS, AAV-iNOS, and Lt- iNOS and their respective control vectors in rat models; 3) to determine efficacy and toxicity of the ideal viral vector (Ad-iNOS, AAV-iNOS, or Lt-iNOS) and its respective control vector as determined in Specific Aim #2, in a porcine model of chronic rejection and 4) to determine the toxicity in human hearts that are being supported with ventricular assist devices. In this proposal we plan to address several key questions including 1) what effect will chronic iNOS expression have on myocardial contractility; 2) will chronic iNOS expression induce cardiac myocyte apoptosis; 3) will iNOS expression induce acute cellular rejection; 4) what effect will iNOS expression during an episode of acute cellular rejection have on global cardiac function and 5) can iNOS be delivered in a safe and efficacious manner such that CAV can be prevented in a large animal model of chronic rejection? We plan to address these questions using both in vitro and in vivo models. Upon completion of the specific alms outlined whin this proposal our goal will be utilize the data obtained from these experiment to serve as a guideline by which to proceed with a safe and efficacious iNOS-based gene therapy trial for the therapeutic prevention of CAV.

根据联合器官共享网络（UNOS）的当前数据，心脏同种异体血管病（CAV）是移植第一年后的主要死亡原因，占第三年后每年死亡的25％。我们实验室的初步研究表明，诱导型一氧化氮合酶（INOS; NOS2）的表达可以抑制新内膜增生的发展并防止CAV的发展。本文所述的提案将检验以下假设：能够转移人INOS基因的病毒载体可以抑制CAV的发展而不会引起对心肌细胞的不适当毒性。此外，该提案将作为未来预防人类CAV的近期临床试验的临床前基础。 In order to serve these ends, we will carry out the following Specific Aims: 1) to determine the efficacy of several viral vectors (adenoviral, adeno-associated virus, and lentivirus) containing the human iNOS gene (Ad-iNOS, AAV-iNOS, Lt-iNOS) and their respective control vectors for suppression of cardiac transplant arteriosclerosis in the rat model; 2）确定大鼠模型中AD-INOS，AAV-INOS和LT- INOS及其各自的对照载体的毒性； 3）确定理想病毒载体（AD-INOS，AAV-INOS或LT-INOS）的功效和毒性及其各自的控制载体，如特定目标2所确定的慢性抑制模型和4）确定在人心脏中受到心脏支持的毒性。在此提案中，我们计划解决几个关键问题，包括1）慢性iNOS表达对心肌收缩性有什么影响； 2）慢性iNOS表达会诱导心肌细胞凋亡； 3）INOS表达会诱导急性细胞排斥； 4）在急性细胞排斥反应的发作中，INOS表达对全球心脏功能有什么影响，而5）INOS可以以安全有效的方式传递，以便在大型的慢性排斥动物模型中可以预防CAV？我们计划使用体外和体内模型解决这些问题。在完成特定施舍的概述后，我们的目标将利用从这些实验中获得的数据作为指导方针，通过该指南，通过该指南进行安全有效的基于INOS的基因治疗试验，以预防CAV的治疗性。