A systematic approach to uncover the basic mechanisms of checkpoint inhibitor immune related adverse events

揭示检查点抑制剂免疫相关不良事件基本机制的系统方法

基本信息

批准号：
10637272
负责人：
Adam Mor
金额：
$ 73.77万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-20 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10637272
关键词：
Address Adverse event Affinity Alleles Animal Model Antibodies Antigens Arthritis Autoimmune Autoimmune Diseases Autoimmunity Automobile Driving Biology Birth Blood CTLA4 blockade Cancer Patient Cell Death Cell Separation Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Clinic Colon Development Disease Disease susceptibility Drug Targeting Evolution Exhibits Failure Future Genetic Genomic DNA Goals Grant Hematopoietic stem cells Heterogeneity Human Immune Evasion Immune checkpoint inhibitor Immune response Immune system Immunofluorescence Immunologic Immunologic Markers Immunophenotyping Immunotherapy Incidence Infiltration Inflammation Inflammatory Inflammatory Response Intervention Joints Life Liver Lung Malignant Neoplasms Mediating Modeling Molecular Mus Musculoskeletal System Organ Pancreas Pathogenesis Pathogenicity Pathway interactions Patients Pattern Peptides Predisposition Process Reaction Role SH2B gene Severities Stains Susceptibility Gene Syndrome Synovial Fluid T cell infiltration T cell receptor repertoire sequencing T-Cell Activation T-Lymphocyte T-Lymphocyte Subsets T-cell receptor repertoire Techniques Testing Thymus Gland Tissues Toxic effect Tumor Antigens Tumor-Infiltrating Lymphocytes Variant Work anti-PD-1 autoreactivity cancer immunotherapy cancer infiltrating T cells cancer therapy checkpoint therapy chemical property chemokine comparative experimental study immune clearance immune-related adverse events inflammatory milieu mouse model neoplastic cell novel peripheral tolerance pre-clinical prevent programmed cell death protein 1 rituximab success therapeutic target tissue injury tocilizumab tool tumor tumor-immune system interactions

项目摘要

ABSTRACT Immune checkpoint inhibitors (ICIs) have profoundly altered cancer treatment. However, despite this technique’s success, there remain major challenges to ICIs use that must be met to best advance these therapies. These include increasing responsiveness to PD-1 and CTLA-4 blockade, uncovering new targets to optimize pathway blockade and addressing why more than one third of all patients that receive ICIs develop often devastating immune related adverse events (irAEs) syndromes characterized by various patterns of off-target organ inflammation. There is currently no clear understanding of the complexity and heterogeneity of irAEs and as ICIs approaches are expanded, understanding the basic mechanisms of irAEs stands to be a critical goal. We hypothesize that three distinguishable, but not mutually exclusive, pathogenic processes drive irAEs, and that the contribution of each varies according to the type of the irAEs. The first process is not tumor-related and is predetermined by known T cell autoimmune susceptibility genes, reflecting a failure of the negative selection mechanism. However, progression to an autoimmune disease is held in a latent preclinical stage by peripheral tolerance mechanisms until triggered by the inhibition of these mechanisms through ICIs. We hypothesize that the second process driving irAEs is an intrinsic consequence of the T cell immune response directed to tumor antigens. We envision that some of the TCR clonotypes responding directly to the tumor are specific for a peptide that is also expressed by target organ MHC molecules involved in the irAEs, and that activation of this expanded clone by ICIs initiates its attack on the tumor and on the target tissue, resulting in irAEs. Finally, since many of the tumor-infiltrating lymphocytes are not clonal, we hypothesize a third process of irAEs wherein the inflammatory response is mediated by promiscuous nonspecific TCR clones activated by ICIs or the tumor inflammatory milieu. Aim 1. To characterize the human T cell repertoire of irAEs patients. We will perform a gDNA repertoire analysis and single cell TCR seq of T cells isolated from the blood, synovial fluid, and synovial tissue of patients with irAEs, to uncover the physical-chemical properties of CDR3s in comparison to T cells that mediate other autoimmune diseases. We will take advantage of the gDNA to test the hypothesis that the entrance of these self-reactive TCRs in a patient’s repertoire is influenced by the variants of the same non-HLA alleles associated with other autoimmune disease susceptibility and the failure of negative selection and examine the role of the MHC allotype on repertoire selection and irAEs development. We will use humanized immune system mice to recreate, from hemopoietic stem cells, a replica of the irAEs patients’ pre-ICIs and pre-tumor TCR repertoire, to study repertoire evolution. Aim 2. To immunophenotype the irAEs mouse model. We developed a novel mouse model for ICIs-induced adverse events. We will perform CITEseq of T cells isolated from tumors and inflamed organs to uncover differential subsets of cells and their functions across different types of tumors. Through tetramer staining and TCRseq, we will uncover the contribution of tumor-specific T cells to the pathogenesis of irAEs. We will utilize multiplex IF studies to uncover the contribution of the tumor and the inflamed organ microenvironment to the function of the infiltrating cells.

抽象的免疫检查点抑制剂（ICIS）具有严重改变的癌症治疗方法。但是，要求这项技术的成功，必须满足ICIS使用的主要挑战，以最好地提高这些疗法。这些包括增加对PD-1和CTLA-4封锁的响应能力，发现新目标以优化途径封锁并解决原因所有接受ICI的患者中有超过三分之一经常出现毁灭性免疫相关的不良事件（IRAE）综合征的特征是各种靶向器官注射的模式。目前没有明确的理解伊拉斯的复杂性和异质性以及随着ICIS方法的扩展，了解基本机制伊拉斯是一个关键目标。我们假设三个独特但不是互斥的致病性过程驱动伊拉斯，并根据伊拉斯的类型对每种品种的贡献。第一个过程不是与肿瘤相关，并由已知的T细胞自身免疫性基因预先确定，反映了阴性的失败选择机制。但是，向自身免疫性疾病的进展是通过周围的潜在临床前阶段的耐受机制，直到通过ICI抑制这些机制触发。我们假设第二驱动IRAE的过程是针对肿瘤抗原的T细胞免疫反应的内在结果。我们设想某些直接响应肿瘤的TCR clonotypes是特异性的，该肽也针对靶标的肽。涉及伊拉斯的器官MHC分子，而ICIS的激活启动了其对侵害的攻击肿瘤和靶组织上，导致伊拉斯。最后，由于许多肿瘤浸润的淋巴细胞不是克隆的我们假设第三个伊拉斯过程，其中炎症反应是由滥交非特异性TCR介导的由ICI或肿瘤炎症环境激活的克隆。目的1。要表征伊拉斯的人类T细胞曲目患者。我们将执行从血液，滑膜分离的T细胞的GDNA曲目分析和单细胞TCR SEQ 与T相比介导其他自身免疫性疾病的细胞。我们将利用GDNA来检验这些自反应性TCR的入口在患者曲目中受到同一非HLA等位基因的变体的影响与其他自身免疫性疾病的敏感性以及负面选择的失败相关，并检查了 MHC关于曲目选择和IRAES开发的同种型。我们将使用人源化免疫系统小鼠重建，从血压干细胞（伊拉斯患者的ICIS和肿瘤前TCR曲目的复制品）到研究曲目进化。目的2。对IRAES小鼠模型进行免疫型。我们开发了一种新型的ICIS诱导的鼠标模型不利事件。我们将执行从肿瘤和发炎器官中分离出的T细胞的Citeseq，以发现差异细胞集及其功能在不同类型的肿瘤上。通过四聚体染色和TCRSEQ，我们将发现肿瘤特异性T细胞对IRAE的发病机理的贡献。如果研究学习，我们将利用多路复用发现肿瘤和发炎器官微环境对浸润细胞功能的贡献。