MEMORIAL SLOAN-KETTERING CANCER GENE THERAPY PROGRAM

在过去三年的赠款期间，MSKCC癌症基因治疗计划项目完全专注于使用逆转录病毒介导的基因转移到造血细胞，目的是(A)改进现有的治疗方案和(B)引入控制恶性疾病的新方法。在过去三年期间取得的主要进展是：在项目中采用了控制恶性疾病的新方法。在过去的三年中，主要的进展是。在项目1中，实现了小鼠和人T淋巴细胞的高效转导；并在体外和体内展示了转导的T淋巴细胞对更昔洛韦的敏感性。在项目2中，实现了树突状细胞的高效转导；并利用它们产生针对卵白蛋白多肽的细胞毒性T细胞。在项目3中，生产新的抗药性酶变体，并在临床前模型中证明，使用转基因造血干细胞涉及到用甲氨蝶呤治疗的荷瘤小鼠的存活。在接下来的5年里，我们计划做以下工作。项目1将完善生产针对EBV淋巴瘤和所有细胞的快速自杀敏感T细胞的方法；将进行针对这些条件的临床试验。项目2将首先在小鼠模型中测试表达黑色素瘤分子的树突状细胞的抗肿瘤效果，然后在III/IV期黑色素瘤患者中进行临床试验。在项目3中，一个新的主要目标将是在非霍奇金淋巴瘤患者中进行临床试验(目前处于IRB批准阶段)，旨在确定表达甲氨蝶呤耐药基因的造血细胞将在体内持续多久，以及药物给药将在多大程度上选择免疫球蛋白分子和表面受体CD28从肿瘤抗原接收共刺激信号。PSMA分子将在人类前列腺癌的临床前模型中作为靶点。逆转录病毒核心将继续支持为所有4个项目开发新的载体，并将在生产用于临床试验的病毒库存方面发挥更大的作用；以及负责持续监测有效性和安全性。行政部门将继续协调所有4个项目，并将在为3个项目的临床试验提供数据管理和为所有人提供生物统计支助方面发挥更大的作用。第三个新的核心正在被引入(成像核心)，这是一种提供最先进技术的成像技术，用于在体内成像以基因工程细胞的肿瘤部位为靶点的体内成像：与项目1和项目4一起开展的工作已经获得了初步数据。项目之间的互动已经广泛且不断增加，特别是因为一个项目所产生的载体设计和转导方案的优化立即使其他项目受益。此外，具有相似目标的调查员和研究员之间的定期联席会议所产生的合作气氛自然对所有项目都产生了宝贵的反馈。这反过来又有助于形成目前的续展提案。

Over the past 3-year grant period, the Program-Project on Cancer Gene Therapy on MSKCC has been entirely focused on the use of retroviral mediated gene transfer into hematopoietic cells for the purpose of (a) improving existing therapeutic protocols and (b) introducing new approaches to the control of malignant diseases. The main advances in this past 3-year period have been: In Project introducing new approaches to the control of malignant diseases. The main advances in this past 3- year period have been. In Project 1, achieving high efficiency transduction of murine and human T lymphocytes; and demonstrating in vitro and in vivo the sensitivity to gancyclovir of the transduced T lymphocytes. In Project 2, achieving high efficiency transduction of dendritic cells; and using them to generate cytotoxic T cells directed against ovalbumin peptides. In Project 3, producing new drug-resistant enzyme variants and demonstrating in a pre-clinical model that the use of genetically modified hematopoietic stem cells involves the survival of tumor-bearing mice treated with methotrexate. Over the next 5 years we plan to do the following. Project 1 will perfect methods for producing rapidly suicide-susceptible T cells targeted to EBV lymphoma and to ALL cells; clinical trials against these conditions will be undertaken. Project 2 will test the anti-tumor efficacy of dendritic cells expressing melanoma molecules first in a mouse model, and then in a clinical trial in patients with stage III/IV melanoma. In Project 3, a new major aim will be a clinical trial (currently at the stage of IRB approval) in patients with non-Hodgkin lymphoma, aiming to establish for how long hematopoietic cells expressing genes conferring resistance to methotrexate will persist in vivo, and to what extent drug administration will select immunoglobulin molecule and the surface receptor CD28 receive a co-stimulator signal from the tumor antigen. The PSMA molecule will be targeted in a pre- clinical model of human prostate carcinoma. The retroviral core will continue to support the development of new vectors for all 4 projects, and it will have a much expanded role in producing viral stocks for use in clinical trials; as well as taking responsibility for continuous monitoring of efficacy and safety. The administrative ore will continue to coordinate all 4 projects, and it will have an expanded role in providing data management for clinical trials in 3 projects and support in biostatistics for all. A third new core is being introduced (Imaging core),a imaging to provide state-of-the-art technology for in vivo imaging of the targeting to tumor sites of genetically engineered cells in mice and eventually in humans: preliminary data have been already obtained in work carried out with Project 1 and Project 4. Interactions among projects have been extensive and increasing particularly because optimization of vector design and transduction protocols accruing form one project have immediately benefitted others. In addition, the atmosphere of collaboration arising from regular joint meetings among investigators and fellows having similar aims has naturally produced valuable feedback on all projects. This in turn has helped in the formation of the present renewal proposal.