GENE THERAPY FOR CANCER

描述：本申请的长期目标是使用基因治疗 在癌症治疗中的应用 在这段时间里， 和新鉴定的细胞凋亡介质将被分析和采用， 小鼠肿瘤缩小的研究。 具体目标按顺序列出 的优先权。 1)为了测试成肌细胞是否能递送FasL， 有效治疗小鼠的实体瘤。 这种方法 利用成肌细胞的属性，使它们能够生长， 大量的，基因工程，并交付给体内组织 稳定表达转基因的细胞 表达FasL的原代成肌细胞， 未转化和Fas缺陷的细胞，将被注射， 测试了它们作为局部抗肿瘤剂的潜力， 通过三种协同机制杀死癌细胞：（a）直接通过 FasL/Fas介导的细胞凋亡，（B）间接通过FasL介导的中性粒细胞 侵袭，和（c）通过旁观者效应作为T和B的同种异体刺激物 淋巴细胞和自然杀伤细胞。 2)为了确定 FasL/Fas介导的细胞凋亡中其他相关蛋白的作用 （a）设计的组成型和可调控的逆转录病毒载体 在申请人的实验室中递送细胞毒性野生型和突变体 以受控的方式将蛋白质递送至大的细胞群体，和（B）a 一种新的方法，用于监测蛋白质-蛋白质相互作用， 基于顺反子内β-半乳糖苷酶（β-gal）的哺乳动物细胞类型 嵌合蛋白的互补。 3)利用贝塔半乳糖 申请人实验室开发的互补技术， 鉴定以前未被识别的相互作用蛋白质， 细胞凋亡，一个潜在的“哺乳动物双杂交筛选。“最终目标是 为了测试Fas介导的细胞杀伤的充分表征的组分， 直接作为抗肿瘤剂并进行鉴定和 描述可能作为未来的途径的其他组成部分 当由成肌细胞或其它体内出现的基因递送时， 交付技术。 结合传统治疗 模式，这些基因治疗策略可以提供有效的治疗方法， 通常难以进入的实体瘤的局部治疗 外科手术和使用抗癌剂的治疗受到阻碍， 剂量限制性骨髓毒性。

DESCRIPTION: The long-term goal of this application is to use gene therapy in the treatment of cancer. In this grant period both well-characterized and newly identified mediators of apoptosis will be analyzed and employed in studies of tumor reduction in mice. The specific aims are listed in order of priority. 1) To test whether delivery of FasL by myoblasts is efficacious in the treatment of solid tumors in mice. This approach capitalizes on the attributes of myoblasts that allow them to be grown to large numbers, genetically engineered, and delivered to tissues in vivo where they stably express transgenes. FasL-expressing primary myoblasts, that are not transformed and are defective in Fas, will be injected and tested for their potential as localized antitumor agents that effectively kill cancer cells by three synergistic mechanisms: (a) directly via FasL/Fas-mediated apoptosis, (b) indirectly via FasL-mediated neutrophil invasion, and (c) via a bystander effect as allogeneic stimulants of T and B lymphocytes and natural killer cells. 2) To determine the mechanism of action of proteins implicated by others in the FasL/Fas mediated apoptotic pathway using (a) constitutive and regulatable retroviral vectors designed in the applicant's laboratory to delivery cytotoxic wild type and mutant proteins in a controlled manner to large populations of cells and (b) a novel method for monitoring protein-protein interactions in diverse intact mammalian cell types based on intracistronic beta-galactosidase (beta-gal) complementation of chimeric proteins. 3) To capitalize on the beta-gal complementation technology developed in the applicant's laboratory to identify previously unrecognized interacting proteins with a function in apoptosis, a potential "mammalian two-hybrid screen." The ultimate goal is to test a well-characterized component of the Fas-mediated cell killing pathway (FasL) directly as an antitumor agent and to identify and characterize other components of the pathway that may serve as future anticancer agents when delivered by myoblasts or other emerging in vivo gene delivery technologies. In conjunction with traditional treatment modalities, these gene therapy strategies may provide potent adjuncts for the localized treatment of solid tumors that are often difficult to access surgically and who treatment with anticancer agents is hindered by dose-limiting bone marrow toxicity.