Reprogramming of T cells for the Treatment of Melanoma

T 细胞重编程治疗黑色素瘤

基本信息

批准号：
8412064
负责人：
Stephen Gottschalk
金额：
$ 101.44万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-13 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8412064
关键词：
Address Adoptive Transfer Affinity Animals Antibodies Antigen Receptors Antigen Targeting Antigens Biochemical Cell Survival Cell physiology Cells Clinical Clinical Investigator Cloning Collaborations Combined Modality Therapy Cytoplasmic Tail Cytotoxic T-Lymphocytes Defect Diagnostic Neoplasm Staging Elements Environment Extracellular Domain Fibroblasts Gene Expression Gene Mutation Genes Genetic Glycolipids Goals Human Immunodeficient Mouse Immunoprecipitation Immunotherapy In complete remission Infiltration Institutes Interleukin-10 Knowledge Lentivirus Vector Lesion Libraries Location Malignant Neoplasms Mass Spectrum Analysis Membrane Proteins Methods Monoclonal Antibodies Mus Mutation Myelogenous Neuroblastoma Organ PTEN gene Pathway interactions Patients Play Population Production Proteins Regulatory T-Lymphocyte Resolution Role Signal Transduction Site Structure Suppressor-Effector T-Lymphocytes Surface Surface Antigens T cell therapy T-Cell Activation T-Lymphocyte T-Lymphocyte Epitopes Testing Therapeutic Tissues Toxic effect Translating Translations Tumor Antigens Work base cancer therapy cell mediated immune response clinical application cytokine cytotoxic density human disease in vivo inhibitor/antagonist interest melanocyte melanoma mouse model mutant neoplastic cell novel oncology peripheral blood programs receptor binding small hairpin RNA small molecule somatic cell nuclear transfer tumor tumor specificity

Address Adoptive Transfer Affinity Animals Antibodies Antigen Receptors Antigen Targeting Antigens Biochemical Cell Survival Cell physiology Cells Clinical Clinical Investigator Cloning Collaborations Combined Modality Therapy Cytoplasmic Tail Cytotoxic T-Lymphocytes Defect Diagnostic Neoplasm Staging Elements Environment Extracellular Domain Fibroblasts Gene Expression Gene Mutation Genes Genetic Glycolipids Goals Human Immunodeficient Mouse Immunoprecipitation Immunotherapy In complete remission Infiltration Institutes Interleukin-10 Knowledge Lentivirus Vector Lesion Libraries Location Malignant Neoplasms Mass Spectrum Analysis Membrane Proteins Methods Monoclonal Antibodies Mus Mutation Myelogenous Neuroblastoma Organ PTEN gene Pathway interactions Patients Play Population Production Proteins Regulatory T-Lymphocyte Resolution Role Signal Transduction Site Structure Suppressor-Effector T-Lymphocytes Surface Surface Antigens T cell therapy T-Cell Activation T-Lymphocyte T-Lymphocyte Epitopes Testing Therapeutic Tissues Toxic effect Translating Translations Tumor Antigens Work base cancer therapy cell mediated immune response clinical application cytokine cytotoxic density human disease in vivo inhibitor/antagonist interest melanocyte melanoma mouse model mutant neoplastic cell novel oncology peripheral blood programs receptor binding small hairpin RNA small molecule somatic cell nuclear transfer tumor tumor specificity

展开

项目摘要

DESCRIPTION (provided by applicant): Recent work has convincingly shown that the degree of T cell infiltration plays a critical role in the natural progression of many cancers. A landmak study found that the type, density, and location of cytotoxic T cells within tumors enabled better prediction of patient survival than histopathological methods currently used to stage cancer. However, the tumor microenvironment strongly inhibits expansion and effector functions of tumor-specific T cells. The goal of this project is to identify novel targes for therapy by pinpointing the key genetic and biochemical defects within tumor-infiltrating T cells that restrain their effector function. The identification of such therapeutic targts is of broad relevance in oncology because T cell mediated immune responses have the potential to eradicate cancers. Towards this goal, we propose a novel in vivo shRN discovery approach that enables identification of critical genes and pathways in the relevant microenvironment. Our hypothesis is that shRNAs which target critical inhibitors within dysfunctional T cells can reprogram them to undergo substantial expansion in tumors. T cells will be genetically modified with shRNA pools and then transferred into tumor-bearing mice so that enrichment of particular shRNAs within tumors can be quantified by Illumina sequencing of the shRNA cassette. This in vivo approach will also allow us to address a second related problem in oncology, the identification of combination therapies that act in a highly synergistic manner on defined cellular pathways. We will approach this issue using a lentiviral vector with two shRNA cloning sites, so that an active shRNA can be tested for synergy against a pool of shRNAs. We will determine which shRNA combinations optimize T cell activityin vivo in terms of proliferation, cytokine production and anti-tumor cytotoxic actio. These therapeutic approaches will be tested in a mouse model in which melanomas spontaneously develop based on genetic lesions found in the human disease. A central goal of this effort is to translate these discoveries into clinical application through collaboration with a clinical investigator with expertise in adoptive T cell therapy. Adoptive transfer of T cells that express 'chimeric antigen receptors' (CARs) has emerged as a promising approach because the antibody-like extracellular domain of a CARbinds with high affinity to a surface molecule on tumor cells, while the cytoplasmic domain induces T cell activation. Co-expression of shRNAs and CARs through the same lentiviral vector into T cells could greatly enhance T cell survival and expansion within tumors. This approach will be tested using human T cells in mice bearing human melanomas, as an important step towards clinical translation. PUBLIC HEALTH RELEVANCE: The goal of this project is to discover novel targets for immunotherapy of melanoma in the relevant in vivo tissue microenvironment using a novel shRNA discovery approach. Such shRNAs may be useful for enhancing the efficacy of adoptive T cell therapy for melanoma. Some of the identified gene products could also be targeted using other approaches, such as small molecules and monoclonal antibodies. Our approach may be widely applicable for the discovery of regulators of T cell function in a variety of other human diseases.

描述（申请人提供）：最近的工作令人信服地表明，T细胞浸润的程度在许多癌症的自然发展中起着至关重要的作用。一项Landmak的研究发现，肿瘤中细胞毒性T细胞的类型，密度和位置比目前用于癌症的组织病理学方法更好地预测了患者生存。然而，肿瘤微环境强烈抑制肿瘤特异性T细胞的膨胀和效应子功能。该项目的目的是通过查明抑制其效应子功能的肿瘤浸润T细胞中的关键遗传和生化缺陷来识别新颖的治疗。这种治疗性TARGT的鉴定在肿瘤学中具有广泛的相关性，因为T细胞介导的免疫反应有可能消除癌症。为了实现这一目标，我们提出了一种新颖的体内发现方法，可以识别相关微环境中的关键基因和途径。我们的假设是，靶向功能障碍T细胞中关键抑制剂的shRNA可以对其进行重新编程以在肿瘤中大量扩张。 t 细胞将用shRNA池进行遗传修饰，然后转移到含肿瘤的小鼠中，以便可以通过 shRNA盒的光明测序。这种体内方法还将使我们能够解决肿瘤学中的第二个相关问题，即对定义的细胞途径以高度协同方式起作用的组合疗法的鉴定。我们将使用带有两个shRNA克隆位点的慢病毒载体来解决此问题，以便可以测试主动shRNA的SHRNA池的协同作用。我们将根据增殖，细胞因子产生和抗肿瘤的细胞毒性ACTIO来确定哪些SHRNA组合优化了T细胞活性素。这些治疗方法将在小鼠模型中进行测试，在小鼠模型中，黑色素瘤根据人类疾病中发现的遗传病变自发发展。这项工作的一个核心目的是通过与具有收养T细胞疗法专业知识的临床研究者合作将这些发现转化为临床应用。表达“嵌合抗原受体”（CARS）的T细胞的过继转移已成为一种有前途的方法，因为碳纤维的抗体样细胞外域对肿瘤细胞上的表面分子高亲和力，而细胞质结构域则诱导T细胞活化。通过相同的慢病毒载体进入T细胞的shRNA和汽车的共表达可以极大地增强T细胞的存活和肿瘤内的扩张。这种方法将使用携带人类黑色素瘤的小鼠中的人T细胞进行测试，这是迈向临床翻译的重要一步。公共卫生相关性：该项目的目的是使用一种新型的ShRNA发现方法来发现相关体内组织微环境中黑色素瘤免疫疗法的新目标。这种SHRNA可能有助于增强收养T细胞疗法对黑色素瘤的功效。一些鉴定的基因产物也可以使用其他方法来靶向，例如小分子和单克隆抗体。我们的方法可能广泛适用于在许多其他人类疾病中发现T细胞功能的调节剂。