Durable CD4+ T-cell Tumor Immunity Following Gene-modified HPSC

基因修饰 HPSC 后的持久 CD4 T 细胞肿瘤免疫

• 批准号: 7735973
• 负责人: Christopher E. Touloukian
• 金额: $ 30万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-22 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7735973
• 关键词: Activated Lymphocyte; Adoptive Transfer; Advanced Malignant Neoplasm; Alleles; Animal Model; Antigens; Applications Grants; Autoantigens; Autoimmune Process; Autoimmunity; CD4 Positive T Lymphocytes; CD8B1 gene; Cancer Vaccines; Cells; Characteristics; Data; Development; Differentiation Antigens; Effectiveness; Engraftment; Event; Experimental Designs; Exposure to; Frequencies; Gene Silencing; Gene-Modified; Goals; HLA-DR4 Antigen; Human; Immune; Immune Tolerance; Immune response; Immune system; Immunization; Immunology; Immunotherapy; Infusion procedures; Interleukin-7; Kinetics; Knock-out; Lead; Leukocytes; Longevity; Lymphocyte; Lymphocyte Activation; Lymphocyte Function; MHC Class II Genes; Malignant Neoplasms; Mediating; Metastatic Melanoma; Modeling; Molecular Genetics; Mus; Patients; Peripheral; Physiological; Play; Population; Positioning Attribute; Proteins; Research Proposals; Role; Running; Self Tolerance; Stem cell transplant; Stem cells; Surface; System; T-Cell Development; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Technology; Testing; Therapeutic; Thymus Gland; Transplantation; Tumor Escape; Tumor Immunity; Tyrosinase related protein-1; Upper arm; Ursidae Family; Vaccination; Vitiligo; base; cancer immunotherapy; cancer therapy; clinical application; cytokine; design; gene therapy; human TYRP1 protein; immunogenicity; improved; innovation; melanocyte; melanoma; novel; novel therapeutics; public health relevance; research study; tumor; tumor eradication

DESCRIPTION (provided by applicant): Over the past 25 years, cancer immunotherapy has undergone tremendous innovation and progress with developments running parallel to advances in basic immunology, molecular genetics and gene therapy. Many of these treatments which have included potent cytokine infusions, the adoptive transfer of activated lymphocytes, and a wide array of cancer vaccines, have been largely applied to patients with metastatic melanoma. These treatments, however, have been consistently hampered by poor immunogenicity, limited immune durability, immune tolerance and tumor escape. To improve upon existing treatments and to help answer basic questions about mechanisms that fundamentally inhibit immune responses to cancer, we sought to develop a novel translational model of cancer immunotherapy. Supported by our preliminary studies, we focused our experimental efforts on further understanding the effects of antigen expression on lymphocyte selection, immune activation, autoimmunity, and cancer treatment. To investigate these issues, we designed a melanoma-based model composed of four critical components: 1) the use of two different experimental mice, one expressing a human class II allele (HLA-DR4), the other knocked-out in the expression of a melanocyte-differentiation antigen called tyrosinase-related-protein-1 (TRP-1); 2) the study of a crucial subset of T-cells (T-helper cells or CD4+ T-cells) which react against TRP-1; 3) the use of lentiviral technology to deliver to hematopoetic stem cells (HPSC) a CD4+ T-cell receptor (TCR) which reacts against TRP-1; and 4) the transplantation and immune reconfiguration with gene-modified HPSC. To achieve these ends we have broken down the overall goal into three specific aims: 1) Does gene-modified HPSC transplantation lead to competent T-cell development and function and can that functionality be further amplified with supplemental IL-7, vaccination, or by early antigen-driven homeostatic proliferation? 2) Does endogenous TRP-1 expression have a direct impact on immunoselection and immune tolerance following gene-modified HPSC transplantation? 3) What are the autoimmune, anti-tumor effects, and mechanisms-of-action of TRP-1-specific CD4+ T-cells following gene-modified HPSC transplantation? We believe this research proposal builds upon existing treatments and technologies in cancer immunotherapy. What makes this model unique however, is both the role of antigen to control the composition of the T-cell repertoire and the critical position that CD4+ T-cells play in global immune activation and anti-tumor activity. We believe that experimental approaches, which facilitate the long-term engraftment, expansion and activation of highly reactive CD4+ T-cells, ultimately produce more effective cancer immunotherapies. PUBLIC HEALTH RELEVANCE: This research proposal builds upon existing technologies and treatments in gene-therapy and immunotherapy for patients with cancer and metastatic melanoma in particular. Here we demonstrate the development of a model in which we successfully reconfigure the immune system with a specific high-frequency white blood cell that is capable of recognizing a key protein found on the surface of melanoma. We then propose experiments designed to better understand the therapeutic implications of this model, improve its overall effectiveness, and study its mechanisms-of-action.

描述（由申请人提供）：在过去的25年中，癌症免疫治疗经历了巨大的创新和进步，其发展与基础免疫学、分子遗传学和基因治疗的进步并行。这些治疗中的许多包括有效的细胞因子输注、活化淋巴细胞的过继转移和广泛的癌症疫苗，已大量应用于转移性黑色素瘤患者。然而，这些治疗一直受到免疫原性差、免疫持久性有限、免疫耐受和肿瘤逃逸的阻碍。为了改进现有的治疗方法，并帮助回答有关从根本上抑制对癌症的免疫反应的机制的基本问题，我们试图开发一种新的癌症免疫治疗的转化模型。在我们初步研究的支持下，我们将实验工作集中在进一步了解抗原表达对淋巴细胞选择、免疫激活、自身免疫和癌症治疗的影响上。为了研究这些问题，我们设计了一个基于黑色素瘤的模型，该模型由四个关键组成部分组成：1）使用两只不同的实验小鼠，一只表达人类II类等位基因（HLA-DR 4），另一只敲除黑色素细胞分化抗原酪氨酸酶相关蛋白-1（TRP-1）的表达; 2）研究T细胞的一个关键子集3）使用慢病毒技术向造血干细胞（HPSC）递送与TRP-1反应的CD4 + T细胞受体（TCR）;（4）基因修饰的HPSC的移植和免疫重构。为了实现这些目标，我们将总体目标分解为三个具体目标：1）基因修饰的HPSC移植是否导致有能力的T细胞发育和功能，以及该功能是否可以通过补充IL-7、疫苗接种或通过早期抗原驱动的稳态增殖进一步扩增？2)内源性TRP-1表达是否对基因修饰的HPSC移植后的免疫选择和免疫耐受有直接影响？3)基因修饰的HPSC移植后TRP-1特异性CD4 + T细胞的自身免疫、抗肿瘤作用和作用机制是什么？我们相信这项研究提案建立在癌症免疫治疗的现有治疗和技术基础上。然而，使该模型独特的是抗原控制T细胞库组成的作用以及CD4 + T细胞在整体免疫激活和抗肿瘤活性中发挥的关键作用。我们认为，实验方法，促进长期植入，扩增和激活高反应性CD4 + T细胞，最终产生更有效的癌症免疫疗法。 公共卫生相关性：这项研究计划建立在基因治疗和免疫治疗的现有技术和治疗方法的基础上，特别是癌症和转移性黑色素瘤患者。在这里，我们展示了一个模型的发展，在这个模型中，我们成功地用一种特定的高频白色血细胞重新配置了免疫系统，这种血细胞能够识别黑色素瘤表面上发现的一种关键蛋白质。然后，我们提出了旨在更好地理解这种模型的治疗意义，提高其整体有效性，并研究其作用机制的实验。