Anti-apoptotic gene therapy: Islet allotransplantation

抗凋亡基因治疗：同种异体胰岛移植

• 批准号: 6398164
• 负责人: Nick Giannoukakis
• 金额: $ 14万
• 依托单位: CHILDREN'S HOSP PITTSBURGH/UPMC HLTH SYS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6398164
• 关键词: CD95 molecule; NOD mouse; SCID mouse; apoptosis; diabetes mellitus therapy; equine infectious anemia virus; gene therapy; immunotherapy; insulin dependent diabetes mellitus; nonhuman therapy evaluation; pancreatic islet transplantation; serine proteinases; technology /technique development; tissue /cell culture; tumor necrosis factor alpha

DESCRIPTION (provided by applicant) Type 1 diabetes mellitus, also termed insulin-dependent diabetes mellitus (IDDM) is an autoimmune disease that specifically targets the pancreatic beta cells of the islets of Langerhans in a T-lymphocyte-mediated destruction. While insulin replacement therapy corrects the hyperglycemia, it is not a cure, since most IDDM patients eventually succumb to the complications associated with imprecise glucose homeostasis. One approach that can restore tight glycemic control is the replacement of beta cells in the form of islet allografts or xenografts. The latter may not become a clinical reality anytime soon primarily because of concerns for zoonoses. Allografts are ethically acceptable, yet they face both alloimmune rejection as well as autoimmune destruction following transplantation. It appears that three distinct death effector pathways are responsible for beta cell destruction: 1) Fas, 2) tumor necrosis factor alpha and 3) perforin/granzyme B. Similar to autoimmune destruction, it appears that Fas, TNFa and perforin/granzyme B are important death effectors in allograft rejection of islets. Genetic engineering of islets to produce inhibitors of these pathways may facilitate allogeneic islet transplantation and may result in long-term survival. In the non-obese diabetic (NOD) mouse model, engineering islets ex vivo to express a variety of immunoregulatory cytokines and proteins has resulted in significant, but not indefinite, prolongation of allogeneic islet transplant survival. One important reason why long-term or indefinite allograft survival has not been achieved is the nature of the gene delivery vectors, most of which are of viral origin and highly immunogenic or toxic to islet cells. Recent engineering of lentiviruses including human immunodeficiency virus (HIV-1), feline immunodeficiency (FIV) and equine infectious anemia viruses (EIAV), has resulted in vectors that are able to infect non-dividing cells, are non-immunogenic in vivo and can stably integrate into the host cell. These desirable characteristics of lentiviral vectors as gene delivery vehicles for islets ex vivo, are in contrast to the highly immunogenic, transient and toxic nature of vectors that have been recently used in islet gene transfer strategies (adenovirus, herpes simplex). While HIV-based lentiviral vectors have been demonstrated to readily infect human islets, the nature of the virus strain is a significant impediment for clinical applications. EIAV, on the other hand, offers the same characteristics as HIV and is not pathogenic in humans. The focus of this proposal is to demonstrate that soluble antagonists of Fas, TNF and granzyme B can protect islets in culture from apoptosis activation and to further develop the EIAV lentiviral system as an efficient gene delivery vector of cDNAs encoding inhibitors of Fas, TNFa and perforin/granzyme B-dependent death effector pathways, alone or in combinations. This may be a desirable approach to facilitate allogeneic islet transplantation as a possible therapy for IDDM.

描述（由申请人提供） 1 型糖尿病，也称为胰岛素依赖型糖尿病 （IDDM）是一种自身免疫性疾病，专门针对胰腺β T 淋巴细胞介导的破坏中的胰岛细胞。尽管 胰岛素替代疗法可以纠正高血糖，但它不是治愈方法，因为 大多数 IDDM 患者最终死于与以下疾病相关的并发症： 不精确的葡萄糖稳态。一种可以恢复紧张血糖的方法 控制是以胰岛同种异体移植物的形式替换 β 细胞或 异种移植物。后者可能不会很快成为临床现实 因为担心人畜共患病。同种异体移植在伦理上是可以接受的，但它们 面临同种免疫排斥以及自身免疫破坏 移植。看来三种不同的死亡效应途径是 负责破坏 β 细胞：1) Fas，2) 肿瘤坏死因子 α 3) 穿孔素/颗粒酶 B。与自身免疫破坏类似，看来 Fas、TNFa 和穿孔素/颗粒酶 B 是同种异体移植物中重要的死亡效应因子 胰岛排斥反应。胰岛基因工程产生抑制剂 这些途径可能促进同种异体胰岛移植并可能导致 在长期生存中。在非肥胖糖尿病（NOD）小鼠模型中，工程 胰岛离体表达多种免疫调节细胞因子和蛋白质，导致同种异体的显着但不是无限期的延长 胰岛移植存活率。长期或无限期的一个重要原因 同种异体移植物的存活尚未实现是基因传递的本质 载体，其中大部分是病毒来源，对人体具有高度免疫原性或毒性 胰岛细胞。包括人类在内的慢病毒的最新工程 免疫缺陷病毒 (HIV-1)、猫免疫缺陷病毒 (FIV) 和马 传染性贫血病毒（EIAV）已产生能够 感染非分裂细胞，体内无免疫原性，可稳定整合 进入宿主细胞。慢病毒载体的这些理想特征是 离体胰岛的基因传递载体与高度 最近使用的载体的免疫原性、瞬时性和毒性 胰岛基因转移策略（腺病毒、单纯疱疹）。虽然基于艾滋病毒 慢病毒载体已被证明可以轻易感染人类胰岛 病毒株的性质是临床研究的重大障碍 应用程序。另一方面，EIAV 具有与 HIV 相同的特征 并且对人类不致病。该提案的重点是证明 Fas、TNF 和颗粒酶 B 的可溶性拮抗剂可以保护胰岛 培养细胞凋亡激活并进一步开发 EIAV 慢病毒 系统作为编码抑制剂的cDNA的有效基因递送载体 Fas、TNFa 和穿孔素/颗粒酶 B 依赖性死亡效应途径，单独或 组合中。这可能是促进同种异体的理想方法 胰岛移植作为 IDDM 的一种可能治疗方法。