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

基本信息

批准号：
10670879
负责人：
Masato Ogishi
金额：
$ 4.77万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
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/10670879
关键词：
Biochemical Biological Assay Biological Markers Biology CD80 gene COVID-19 pneumonia 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 Malignant Neoplasms Mediating Methods Minor Minority Mismatch Repair Mismatch Repair Deficiency Modeling Molecular Mus Mutation Neoplasm Metastasis Outcome Pathway interactions Patients Predictive Value RNA Interference Reporting Reproducibility Research Research Personnel Role Self Tolerance Side Signal Transduction Somatic Mutation Sorting Surface T cell response T-Cell Activation T-Lymphocyte Testing Therapeutic Tumor Immunity Uncertainty Work cancer cell cancer immunotherapy cancer type candidate validation 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

项目摘要

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人类 T 细胞和癌细胞之间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。