ICAM1 gene inactivation protects against ischemic injury

所有实体器官同种异体移植物都在体外维持一段时间。 虽然不受欢迎的缺血性损伤的结果从这个离体阶段，它确实提供了一个独特的窗口，集中和有效的基因转移，可以提高移植物和受体的生存。 移植物功能延迟恢复和原发性无功能仍然是移植中难以解决的问题，并与慢性排斥反应引起的移植物晚期衰竭有关。 缺血/再灌注损伤（IRI）是一个重要的抗原非依赖性因素，可以有助于这些临床实体的发病机制。 我们已经开发了一种嵌合表达载体，该载体在原位产生丰富的反义/核酶调节分子，并已被证明有效。 这项工作的目标是建立条件，通过有效的离体递送我们的嵌合转基因到同种异体移植物中的个别基因的选择性失活负责IRI。 粘附分子ICAM-1已被选为模型靶基因，以开发这种基因治疗策略，因为有令人信服的证据表明，它是一个重要的触发分子的级联反应的促炎介质负责IRI。 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。 使用合理的方法来设计嵌合构建体并在体外对其进行测试，我们希望建立在移植的同种异体移植物中抑制ICAM-1和IRI通路中的其他重要分子的要求和条件。 将开发用于大鼠肾IRI模型的递送有效ICAM-1抑制转基因的病毒和非病毒方法。 由于这种抑制方法是基于比以前采用的策略更有效和特异的技术，因此它具有作为强大的治疗剂和询问基因功能的替代遗传工具的巨大潜力。 这项工作具有重要意义的选择性失活的基因功能在同种异体移植物通过体外基因转移。

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.