ICAM1 gene inactivation protects against ischemic injury

ICAM1 基因失活可预防缺血性损伤

• 批准号: 6881422
• 负责人: ROBERT A MONTGOMERY
• 金额: $ 37.96万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6881422
• 关键词: adeno associated virus group; cell adhesion molecules; complementary RNA; gene expression; genetic models; immunocytochemistry; inflammation; injury; ischemia; kidney function; laboratory mouse; laboratory rat; leukocyte adhesion molecules; messenger RNA; model design /development; northern blottings; polymerase chain reaction; regulatory gene; renal ischemia /hypoxia; reperfusion; ribozymes; small nuclear RNA; tissue /cell culture; transfection /expression vector; vascular endothelium

All solid organ allografts are maintained for some period of time outside of the body. While unwelcome ischemic injury results from this ex vivo phase, it does offer a unique window for focused and efficient gene transfer that could improve graft and recipient survival. Delayed graft function and primary nonfunction remain recalcitrant problems in transplantation and have been associated with late graft failure due to chronic rejection. Ischemia/reperfusion injury (IRI) is an important antigen-independent factor that can contribute to the pathogenesis of these clinical entities. We have developed a chimeric expression vector that generates abundant antisense/ribozyme regulatory molecules in situ and has proven efficacy. The goal of this work is to establish the conditions for the selective inactivation of individual genes responsible for IRI through the efficient ex vivo delivery of our chimeric transgene into allografts. The adhesion molecule ICAM-1 has been chosen as a model target gene to develop this gene therapy strategy because there is compelling evidence that it is an important trigger molecule for cascades of proinflamatory mediators responsible for IRI. Our hypothesis is that the ex vivo delivery of a chimeric transgene designed to produce anti- ICAM-1 regulatory molecules will inhibit the expression of ICAM-I in endothelial cells and abrogate or ameliorate IRI in an ischemic organ. Using a rational approach to designing chimeric constructs and testing them in vitro, we hope to establish the requirements and conditions for inhibition of ICAM-1 and other important molecules in the IRI pathway in transplanted allografts. Both viral and nonviral approaches to delivering effective ICAM-1 inhibitory transgenes will be developed for use in a rat renal IRI model. Because this method of inhibition is based on technology that is more potent and specific than strategies previously employed, it possess great potential as both a powerful therapeutic agent and a surrogate genetic tool for interrogating gene function. This work has important implications for the selective inactivation of gene function in allografts through ex vivo gene transfer.

所有实体器官同种异体移植物都会在体外维持一段时间。 虽然这种离体阶段会导致不受欢迎的缺血性损伤，但它确实为集中有效的基因转移提供了一个独特的窗口，可以提高移植物和受体的存活率。 移植物功能延迟和原发性无功能仍然是移植中的顽固问题，并且与慢性排斥导致的晚期移植物失败有关。 缺血/再灌注损伤（IRI）是一个重要的抗原非依赖性因素，可能导致这些临床疾病的发病机制。 我们开发了一种嵌合表达载体，可原位产生丰富的反义/核酶调节分子，并已被证明有效。 这项工作的目标是通过将我们的嵌合转基因有效离体递送至同种异体移植物中，建立选择性失活负责 IRI 的单个基因的条件。 粘附分子 ICAM-1 已被选为开发这种基因治疗策略的模型靶基因，因为有令人信服的证据表明它是负责 IRI 的促炎介质级联的重要触发分子。 我们的假设是，设计用于产生抗ICAM-1调节分子的嵌合转基因的离体递送将抑制内皮细胞中ICAM-1的表达并消除或改善缺血器官中的IRI。 使用合理的方法设计嵌合结构并在体外测试它们，我们希望建立在移植的同种异体移植物中抑制 ICAM-1 和 IRI 途径中其他重要分子的要求和条件。 将开发用于传递有效 ICAM-1 抑制性转基因的病毒和非病毒方法，用于大鼠肾 IRI 模型。 由于这种抑制方法基于比以前采用的策略更有效和更具体的技术，因此它具有作为强大的治疗剂和用于询问基因功能的替代遗传工具的巨大潜力。 这项工作对于通过离体基因转移选择性失活同种异体移植物中的基因功能具有重要意义。