GENE THERAPY FOR CANCER

描述：该应用程序的长期目标是使用基因疗法 在治疗癌症方面。在这一批款期内， 新发现的细胞凋亡介体将被分析并应用于 对小鼠肿瘤抑制作用的研究。具体目标按顺序列出 最重要的是。1)检测成肌细胞传递FasL是否 对小鼠实体瘤有明显的治疗作用。这种方法 利用成肌细胞的特性，使它们能够培养成 大量的，基因工程的，并被输送到体内组织中 在那里它们稳定地表达转基因。表达FasL的原代成肌细胞， 没有转化和在FAS中有缺陷的，将被注射和 测试了它们作为局部抗肿瘤药物的潜力，有效地 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，以及(C)通过旁观者效应作为同种异体T和B的刺激物 淋巴细胞和自然杀伤细胞。2)确定其作用机制 其他相关蛋白在FasL/Fas介导的细胞凋亡中的作用 使用(A)构造性和可调节的逆转录病毒载体的途径 在申请人的实验室中交付细胞毒性野生型和突变体 以受控的方式将蛋白质传递给大量细胞和(B)a 一种监测不同物种蛋白质相互作用的新方法 基于β-半乳糖苷酶(β-GAL)的哺乳动物细胞类型 嵌合蛋白的互补作用。3)利用Beta-Gal 申请人实验室开发的补充技术 鉴定以前未识别的相互作用蛋白，其功能在 细胞凋亡，一个潜在的“哺乳动物双杂交屏幕”。最终目标是 测试Fas介导的细胞杀伤的一个特征良好的成分 直接作为抗肿瘤药物的途径(FasL)并鉴定和 描述未来可能发挥作用的路径的其他组成部分 成肌细胞或体内其他新兴基因携带的抗癌药物 交付技术。与传统治疗相结合 这些基因治疗策略可能提供有效的辅助手段 对经常难以触及的实体肿瘤的局部治疗 手术和世卫组织使用抗癌药物治疗受到以下因素的阻碍 剂量限制的骨髓毒性。

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.