Modulation of Cell Death during Cytotoxic Gene Therapy

细胞毒性基因治疗过程中细胞死亡的调节

• 批准号: 6414786
• 负责人: Richard G. Vile
• 金额: $ 25.72万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6414786
• 关键词: Adenoviridae; antigen antibody reaction; biological signal transduction; biotechnology; cell death; cytokine; cytotoxicity; flow cytometry; gene delivery system; gene expression; gene targeting; gene therapy; heat shock proteins; immunogenetics; immunologic assay /test; laboratory mouse; leukocyte activation /transformation; macrophage; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; neoplastic cell; nonhuman therapy evaluation; nucleic acid hybridization; stress proteins; technology /technique development; transcription factor; transfection /expression vector; tumor antigens

The mechanisms by which the immune system distinguishes normal developmental cell death from pathological immunogenic cell killing are central to effective cancer immunotherapy. The goal of this project is to understand the mechanisms by which tumor cell death can induce immune activation, and exploit these mechanisms to develop novel cytotoxic gene therapies for cancer. Preliminary data indicates that the mode of cell death is critical to generation of effective anti-tumor immune responses, such that cells dying via non-apoptotic mechanisms generate greater immune activation than cells dying via apoptotic mechanisms. We have also shown that macrophages are a key cell distinguishing between tumor cells dying through classical apoptosis and cells engineered to die through non-apoptotic mechanisms, and these differential responses of macrophages can be used to cure mice of established tumors. These data indicate that there are specific characteristics of non-apoptotic cell killing that result in immune activation. We aim to develop novel cytotoxic gene therapies that combine the key features of non-apoptotic cell killing, including release of cell contents, activation of cellular stress programs, and generation of an inflammatory tumor environment. To achieve this aim, in combination with candidate cytotoxic genes we will develop a novel approach in gene therapy that involves delivery of transcription factors and signaling molecules to activate broad- spectrum cellular gene expression programs. In view of preliminary data indicating that expression of cell stress- related genes such as Hsp70 by tumor cells during cell killing is key to immune activation, we aim to activate, by genetic means, programs of cell stress in tumor cells. Additionally in view of data indicating that immune activation is achieved in a pro- inflammatory cytokine environment generated by macrophages exposed to non-apoptotic tumor death, concomitant with cell death we aim to activate the broad immune cascade characteristic of the anti-viral response. We hypothesize that this combination of non-apoptotic cell death in a manner and an environment optimal for immune activation will result in increased efficacy of cytotoxic gene therapy approaches. Thus through increased understanding the mechanisms by which immune processes are activated by cytotoxic modalities we will significantly overcome some of the major current limitations of cancer gene therapy; namely limited efficiency of gene delivery and limited efficacy of in vivo cellular cytotoxicity.

免疫系统将正常发育细胞死亡与病理免疫原性杀害区分开的机制对于有效的癌症免疫疗法至关重要。 该项目的目的是了解肿瘤细胞死亡可以诱导免疫激活的机制，并利用这些机制来开发新的细胞毒性基因疗法。初步数据表明，细胞死亡模式对有效的抗肿瘤免疫反应的产生至关重要，因此，通过非凋亡机制死亡的细胞比通过凋亡机制死亡的细胞产生更大的免疫激活。 我们还表明，巨噬细胞是通过经典细胞凋亡而垂死的肿瘤细胞和通过非凋亡机制死亡的细胞的关键细胞，并且这些巨噬细胞的差异反应可用于治愈已建立肿瘤的小鼠。 这些数据表明，非凋亡细胞杀伤的特定特征会导致免疫激活。 我们旨在开发新型的细胞毒性基因疗法，以结合非凋亡细胞杀伤的关键特征，包括细胞含量的释放，细胞应激程序的激活以及产生炎症性肿瘤环境。 为了实现这一目标，与候选细胞毒性基因相结合，我们将开发一种基因疗法的新方法，涉及转录因子和信号分子的传递，以激活广泛的细胞基因表达程序。 鉴于初步数据表明，在细胞杀死过程中，肿瘤细胞的细胞应激相关基因（例如HSP70）的表达是免疫激活的关键，我们旨在通过遗传手段激活肿瘤细胞中细胞应激的程序。 此外，鉴于数据表明在暴露于非凋亡肿瘤死亡的巨噬细胞产生的炎性细胞因子环境中实现了免疫活化，与细胞死亡有关，我们旨在激活抗病毒反应的广泛免疫级联反应特征。 我们假设以某种方式与非凋亡细胞死亡和免疫激活最佳环境的这种结合将导致细胞毒性基因治疗方法的疗效提高。 因此，通过增加了解免疫过程被细胞毒性形态激活的机制，我们将显着克服癌症基因治疗的一些主要当前局限性。即，基因递送效率有限，体内细胞毒性的功效有限。