Genetic and immunological dissection of coinhibitory crosstalks between human T cells and cancer cells

人类 T 细胞和癌细胞之间共抑制串扰的遗传和免疫学剖析

• 批准号: 10529610
• 负责人: Masato Ogishi
• 金额: $ 4.83万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10529610
• 关键词: Biochemical; Biological Assay; Biological Markers; Biology; CD80 gene; CRISPR screen; Cancer Patient; Cancer cell line; Cell Line; Cells; Child; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Communicable Diseases; Development; Disease remission; Dissection; Etiology; Favorable Clinical Outcome; Functional disorder; Genes; Genetic; Germ-Line Mutation; Goals; Growth; Health; Human; Human Genetics; Immobilization; Immune; Immunity; Immunologics; Immunooncology; Immunosuppression; Immunotherapy; In Vitro; Inherited; Interferon Type I; Intrinsic factor; Knock-out; Lesion; Ligands; Longitudinal Studies; Malignant Neoplasms; Mediating; Methods; Minor; Minority; Mismatch Repair; Modeling; Molecular; Mus; Mutation; Neoplasm Metastasis; Outcome; Pathway interactions; Patients; Predictive Value; RNA Interference; Reporting; Research Personnel; Role; Self Tolerance; Side; Signal Transduction; Somatic Mutation; Sorting - Cell Movement; Surface; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Tumor Immunity; Uncertainty; Work; cancer cell; cancer immunotherapy; cancer type; cytotoxicity; genetic approach; genome-wide; immune cell checkpoints; immune checkpoint; immune checkpoint blockade; immunoregulation; improved; inter-individual variation; neoantigens; neoplastic cell; next generation; novel; patient subsets; pembrolizumab; programmed cell death ligand 1; programmed cell death protein 1; receptor; response; severe COVID-19; success; systemic autoimmunity; transcriptome sequencing; tumor; tumor immunology

PROJECT SUMMARY / ABSTRACT The goal of this proposal is to identify novel coinhibitory checkpoint molecules operating orthogonally to currently targeted checkpoints in cancer patients, such as PD-1. Despite the unprecedented success of checkpoint blockade immunotherapy as a therapeutic approach against multiple types of cancer, significant inter-individual variability remains, ranging from no response to complete remission of systemically metastasized lesions. Various tumor-intrinsic parameters, such as tumor mutational burden, neoantigen burden, and mismatch repair pathway deficiencies, only show limited predictive power for favorable clinical outcomes. Therefore, complementary to such tumor-intrinsic factors, it is plausible to hypothesize that humans have hitherto unknown coinhibitory mechanisms orthogonal to currently known immune checkpoints and that a minor fraction of humans with germline or somatic mutations in such pathways could benefit from exceptionally potent antitumor immunity triggered by checkpoint blockade immunotherapy. Identification of such orthogonal pathways would significantly enhance the health of patients with multiple types of cancer; for instance, the 15 types of cancer for which pembrolizumab has been indicated. The current project aims at dissecting the human genetic and immunological basis of coinhibitory crosstalk between human T cells and cancer cells by tackling the two specific aims. Specific Aim 1 is to delineate the molecular mechanisms of PD-L1/L2-mediated inhibition of T cell activation independent from PD-1. This aim is supported by preliminary observations that both primary and immortalized T cells from a patient with inherited complete PD-1 deficiency, as well as PD-1-negative human T cell lines Jurkat and HuT78, all responded to co-inhibition by bead- immobilized PD-L1/L2. CD83 was identified as a candidate for PD-L1-mediated immunosuppression. The proposed work includes searching for additional candidates through RNA sequencing and surface receptor profiling and functionally validating the candidates via RNA silencing and CRISPR-guided knockout. Specific Aim 2 is to characterize the immunoregulatory function of CD83 in human T cells. Previous studies suggest that CD83 can serve as both an immunosuppressive receptor and ligand for human T cells, although its counterreceptor(s), downstream signaling, and relevance to antitumor immunity remain mostly elusive. The proposed work includes biochemical characterization of CD83 signaling in human T cells with or without cancer cell coculture utilizing genetic knockout and lentiviral rescue of CD83 gene. The proposed project will help untangle the biochemical and immunological basis of human T cell coinhibitory circuits in the crosstalk between cancer cells and also substantially bolster the applicant's scientific portfolio and expedite the growth toward an independent investigator pioneering human genetics of cancer immunology and immunotherapy.

项目概要/摘要 该提案的目标是识别正交操作的新型共抑制检查点分子 目前癌症患者的目标检查点，例如 PD-1。尽管取得了前所未有的成功 检查点阻断免疫疗法作为多种癌症的治疗方法， 仍然存在显着的个体差异，从无反应到完全缓解 全身转移病灶。各种肿瘤内在参数，例如肿瘤突变 负担、新抗原负担和错配修复途径缺陷仅显示有限的预测 良好的临床结果的力量。因此，与这些肿瘤内在因素相辅相成的是 有理由假设人类具有迄今为止未知的正交共抑制机制 目前已知的免疫检查点，并且一小部分人患有种系或体细胞 这些途径的突变可能会受益于由以下因素引发的异常有效的抗肿瘤免疫： 检查点阻断免疫疗法。识别这种正交路径将显着 增强多种癌症患者的健康；例如，15种癌症 已表明派姆单抗。当前的项目旨在剖析人类遗传 通过解决人类 T 细胞和癌细胞之间的共抑制串扰的免疫学基础 两个具体目标。具体目标 1 是描述 PD-L1/L2 介导的分子机制 独立于 PD-1 的 T 细胞活化抑制。这一目标得到了初步观察的支持 来自患有遗传性完全 PD-1 缺陷的患者的原代 T 细胞和永生化 T 细胞，如 以及 PD-1 阴性人类 T 细胞系 Jurkat 和 HuT78，均对珠子的共抑制做出反应 固定化 PD-L1/L2。 CD83 被确定为 PD-L1 介导的免疫抑制的候选者。 拟议的工作包括通过 RNA 测序和表面寻找其他候选者 通过 RNA 沉默和 CRISPR 引导对候选药物进行受体分析和功能验证 昏死。具体目标 2 是表征人类 T 细胞中 CD83 的免疫调节功能。 先前的研究表明 CD83 可以作为免疫抑制受体和配体 人类 T 细胞，尽管其反受体、下游信号传导以及与抗肿瘤的相关性 免疫力仍然难以捉摸。拟议的工作包括 CD83 的生化表征 利用基因敲除和慢病毒在有或没有癌细胞共培养的人类 T 细胞中进行信号传导 CD83基因的拯救。拟议的项目将有助于解开生化和免疫学基础 人类 T 细胞共抑制回路在癌细胞之间的串扰中的作用，并且还大大增强了 申请人的科学组合并加速成长为独立研究者 开创了癌症免疫学和免疫治疗的人类遗传学。