Conditional Regulation of T Cell Survival in Immunotherapy

免疫治疗中 T 细胞存活的条件调节

• 批准号: 7154577
• 负责人: STANLEY R. RIDDELL
• 金额: $ 38.21万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-29 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7154577
• 关键词: CD34 molecule; Retroviridae; T lymphocyte; apoptosis; biotechnology; blood /lymphatic neoplasm; clinical research; clinical trials; cooperative study; cysteine endopeptidases; dimer; gene induction /repression; gene therapy; graft versus host disease; hematopoietic stem cells; human subject; human therapy evaluation; immunogenetics; immunotherapy; neoplasm /cancer relapse /recurrence; passive immunization; patient oriented research; small molecule; stem cell transplantation; tissue /cell culture; transfection; transfection /expression vector

Small molecules that control the expression and/or activity of transgene products can be used to initiate signaling pathways that regulate cellular functions, including cell death. Studies supported by this grant have investigated inducible suicide genes based on the induction of apoptosis through the chemical induced dimerization of FK506 binding proteins (FKBP) linked to death effector molecules. This strategy for regulating cell survival employs transgenes that encode human proteins and should avoid the problem of immune responses to suicide gene products derived from pathogens. A new construct that utilizes an inducible caspase-9 gene (iCasp-9) for cell suicide and a truncated CD34 molecule for selection of transduced cells has been developed to overcome the limitations of the Fas suicide gene studied in the prior funding period. The proposed studies will evaluate retroviral vectors encoding the iCasp-9 suicide gene in vitro and in a clinical trial to regulate allogeneic donor T lymphocytes administered to patients to treat recurrent or persistent malignancy after allogeneic hematopoietic stem cell transplant (HSCT). The introduction of iCasp-9 will bring the T cells under pharmacologic control and permit their ablation in patients with GVHD. These studies will provide insight into the clinical use of chemical dimerizing agents to regulate transgene function and may identify a suicide gene that is not immunogenic in humans and could be broadly applied in cell and gene therapy. The specific aims are: 1. To determine the in vitro activity of retroviral constructs encoding truncated CD34 as a selectable marker and inducible caspase-9 (iCasp-9) for cell suicide. 2. To determine the safety, in vivo persistence, and biologic activity of adoptive immunotherapy with donor T lymphocytes modified by retrovirus mediated gene transfer to express the ACD34-iCasp-9 suicide gene. 3. To determine if donor T lymphocytes modified to express the ACD34/iCasp-9 suicide gene can be ablated in patients who develop graft versus host disease or other serious toxicity by the administration of the synthetic FKBP binding drug AP1903.

控制转基因产物的表达和/或活性的小分子可用于启动调节细胞功能（包括细胞死亡）的信号传导途径。该基金支持的研究调查了诱导型自杀基因，其基于通过与死亡效应分子连接的FK 506结合蛋白（FKBP）的化学诱导二聚化诱导细胞凋亡。这种调节细胞存活的策略采用编码人类蛋白质的转基因，并且应该避免对来自病原体的自杀基因产物的免疫应答的问题。开发了一种新的构建体，该构建体利用诱导型半胱天冬酶-9基因（iCasp-9）进行细胞自杀，并利用截短的CD 34分子选择转导的细胞，以克服先前资助中研究的Fas自杀基因的局限性。 期拟议的研究将在体外和临床试验中评估编码iCasp-9自杀基因的逆转录病毒载体，以调节给予患者的同种异体供体T淋巴细胞，以治疗同种异体造血干细胞移植（HSCT）后复发或持续的恶性肿瘤。iCasp-9的引入将使T细胞处于药理学控制之下，并允许在GVHD患者中进行消融。这些研究将提供深入了解化学二聚化剂的临床应用，以调节转基因功能，并可能确定一个自杀基因，在人类中是没有免疫原性的，可以广泛应用于细胞和基因治疗。具体目标是：1.确定编码截短的CD 34（作为选择性标记）和诱导型胱天蛋白酶-9（iCasp-9）的逆转录病毒构建体的体外细胞自杀活性。2.确定供者过继免疫治疗的安全性、体内持久性和生物活性 通过逆转录病毒介导的基因转移修饰以表达ACD 34-iCasp-9自杀基因的T淋巴细胞。3.确定在发生移植物抗宿主病或其他严重毒性的患者中，是否可以通过给予合成FKBP结合药物AP 1903来消融经修饰以表达ACD 34/iCasp-9自杀基因的供体T淋巴细胞。