MEMORIAL SLOAN-KETTERING CANCER GENE THERAPY PROGRAM

在过去3年的资助期内，MSKCC癌症基因治疗方案项目完全侧重于利用逆转录病毒介导的基因转移到造血细胞中，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。过去3年期间的主要进展是：在项目中引入了控制恶性疾病的新方法。过去三年的主要进展是。项目1：实现小鼠和人T淋巴细胞的高效转导；并在体外和体内证明了转导的T淋巴细胞对更昔洛韦的敏感性。项目2：实现树突状细胞的高效转导；并利用它们产生针对卵清蛋白肽的细胞毒性T细胞。在项目3中，产生新的耐药酶变体，并在临床前模型中证明，使用基因修饰的造血干细胞涉及接受甲氨蝶呤治疗的荷瘤小鼠的存活。在接下来的5年里，我们计划做以下事情。项目1将完善生产针对EBV淋巴瘤和ALL细胞的快速自杀敏感T细胞的方法；针对这些情况将进行临床试验。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功效，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在体内持续多长时间，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接收来自肿瘤抗原的共刺激信号。PSMA分子将成为人类前列腺癌临床前模型的靶标。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；同时负责持续监测药物的有效性和安全性。行政部门将继续协调所有4个项目，并在为3个项目的临床试验提供数据管理和为所有项目提供生物统计支持方面发挥更大的作用。正在引入第三个新核心（成像核心），这是一种成像技术，可提供最先进的技术，用于在小鼠和最终在人类中针对基因工程细胞的肿瘤部位进行体内成像：在项目1和项目4的工作中已经获得了初步数据。项目之间的相互作用已经广泛并不断增加，特别是因为一个项目产生的载体设计和转导协议的优化立即使其他项目受益。此外，具有类似目标的研究人员和研究员之间定期举行联合会议所产生的合作气氛自然对所有项目产生了宝贵的反馈。这反过来又有助于形成目前的更新建议。

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.