CDK4/6 inhibition during CD8 T cell priming potentiates memory formation in mice and humans

CD8 T 细胞启动期间的 CDK4/6 抑制可增强小鼠和人类的记忆形成

基本信息

批准号：
10382325
负责人：
Stephanie Dougan
金额：
$ 44.5万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-05 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10382325
关键词：
ATAC-seq Achievement Adjuvant Adoptive Transfer Affect Affinity Antigens Binding Binding Sites Blood Blood Cells Blood Circulation Blood specimen Breast Cancer Patient Breast Cancer Treatment CD8-Positive T-Lymphocytes CD8B1 gene CDK4 gene Cancer Patient Cell Cycle Cell Cycle Regulation Cell Lineage Cell division Cells Chromatin Clinic Clinical Clinical Trials Clinical Trials Design Co-Immunoprecipitations Communicable Diseases Cytostatics DNA Data Destinations Development Disease remission Effector Cell G1 Arrest Gene Expression Gene Expression Profile Gene Expression Profiling Gene Silencing Genes Genetic Transcription Goals Hepatitis Heterogeneity Human Immunity Immunologics Immunotherapeutic agent In Vitro Individual Link Malignant Neoplasms Mass Spectrum Analysis Memory Minority Modeling Molecular Mus Nature PD-1 blockade Paper Patients Pharmaceutical Preparations Physiological Positioning Attribute Proteins RNA RUNX3 gene Reporting Resolution Role Sampling Seminal T cell therapy T memory cell T-Lymphocyte Technology Testing Tissue Sample Tissues Toxic effect Transcription Repressor Tumor Immunity Up-Regulation blood treatment cancer immunotherapy chromatin immunoprecipitation cytokine drug action immune checkpoint blockade improved in vivo inhibitor inhibitor therapy long term memory mouse model neoplastic cell patient safety phase I trial phase II trial prevent response therapy design transcription factor tumor tumor growth

项目摘要

Project Summary Cancer immunotherapy’s signature achievement is extension of overall survival with long-term durable remissions in a minority of patients. However, the question of how memory CD8 T cells form in cancer patients is poorly understood. Our long-term goal is to understand the relationship between cell cycle regulation and CD8 T cell memory fate. We examined CDK4/6 inhibition in mouse and human CD8 T cells during T cell priming and found a dramatic skewing of CD8 T cells toward a memory fate. Antigen-specific CD8 T cells treated ex vivo with CDK4/6i displayed increased long-lived memory responses upon adoptive transfer into mice. We further used single cell transcriptional profiling to evaluate recently activated human CD8 T cells from blood of breast cancer patients pre- and on-treatment with the CDK4/6i palbociclib or abemaciclib. TCR clonotype tracking and RNA velocity analysis revealed that CDK4/6 inhibition increased the ratio of central memory to effector CD8 T cell precursors. Analysis of paired samples showed that MYC targets were downregulated by CDK4/6 inhibitor therapy in humans, consistent with an increase in the transcriptional repressor MXD4. Our central hypothesis is that CDK4/6 inhibition in CD8 T cells in both mice and humans augments long-term protective immunity. To test this, we will use state-of-the-art technologies including: 1) TRP1 transnuclear mice developed by our lab to study antigen-specific TCRs with a physiologic range of affinities for an endogenous antigen; 2) scRNAseq and TCR clonotype tracking to analyze a large bank of longitudinal blood and matched tissue samples from patients on a Phase II trial; 3) ChIPseq, and co-immunoprecipitations to determine the molecular basis linking CDK4/6 inhibition to memory cell fate in CD8 T cells. In Aim 1 we will identify the molecular mechanism linking CDK4 and/or CDK6 to induction of memory cell fate. Slowing the cell cycle using other cell cycle inhibitors or by taking advantage of natural heterogeneity in cell division rates does not potentiate memory formation in CD8 T cells, implying a unique role for CDK4 and/or CDK6 in conferring a memory cell fate. We will identify the transcriptional regulators affected by CDK4/6 inhibition, with a focus on MYC and RUNX3. Combination of CDK4/6 inhibitors with PD-1 blockade in the clinic results in unexpected treatment-limiting toxicity. In Aim 2 we propose that a short course of CDK4/6 inhibition would successfully potentiate CD8 T cell memory, thus allowing cessation of the CDK4/6 inhibitor to minimize or obviate overlap with PD-1 blockade. We will test this hypothesis in mice using our TRP1 CD8 T cell adoptive transfer model, and in humans using 29 paired blood and tissue samples from a Phase I trial of CDK4/6 inhibition with PD-1 blockade. We will use high- resolution single-cell transcriptional profiling of recently activated CD8 T cells to track individual TCR clonotypes and their transcriptional profiles in CDK4/6 inhibitor pre- and on-treatment blood and tissue samples. Impact: A basic understanding of the mechanistic role of CD4/6 in CD8 T cell fate decisions will allow for better design of clinical trials in cancer or infectious disease using CDK4/6i as adjuvants to promote CD8 T cell memory.

项目摘要癌症免疫疗法的签名成就是延长整体生存期并长期耐用性少数患者的减免。但是，癌症患者中记忆CD8 T细胞如何形成的问题理解很差。我们的长期目标是了解细胞周期调节之间的关系和CD8 T细胞存储器命运。我们检查了T细胞中小鼠和人CD8 T细胞中的CDK4/6抑制启动并发现CD8 T细胞朝着记忆命运的戏剧性偏斜。处理过的抗原特异性CD8 T细胞带有CDK4/6i的外体显示在自适应转移到小鼠时增加了长寿命的记忆反应。我们进一步使用单细胞转录分析来评估最近激活的人CD8 T细胞乳腺癌患者与CDK4/6i palbociclib或Abemaciclib进行治疗前和治疗。 TCR克隆型跟踪和RNA速度分析表明，CDK4/6抑制增加了中央记忆与效应子CD8 T细胞前体。对配对样品的分析表明，MYC目标被下调人类的CDK4/6抑制剂疗法与转录复制MXD4的增加一致。我们的中心假设是小鼠和人类CDK4 T细胞中CDK4/6抑制作用长期增强保护性免疫。为了测试这一点，我们将使用最新技术，包括：1）TRP1 TRENNACOIL MICE 由我们的实验室开发，用于研究具有内生的生理范围的抗原特异性TCR 抗原; 2）SCRNASEQ和TCR克隆型跟踪，分析一大群纵向血液并匹配 II期试验中患者的组织样品； 3）chipseq和共免疫沉淀以确定将CDK4/6抑制与CD8 T细胞中的记忆细胞命运联系起来的分子基础。在AIM 1中，我们将确定将CDK4和/或CDK6连接到记忆细胞命运的分子机制。使用使用其他细胞周期抑制剂或通过利用细胞分裂率的自然异质性没有潜力 CD8 T细胞中的记忆形成，这意味着CDK4和/或CDK6的独特作用在会议记忆细胞命运中。我们将确定受CDK4/6抑制影响的转录调节器，重点是MYC和RUNX3。 CDK4/6抑制剂与诊所中PD-1阻滞的组合导致意外的治疗限制毒性。在AIM 2中，我们建议简短的CDK4/6抑制作用将成功潜在的CD8 T细胞记忆，从而允许停止CDK4/6抑制剂最大程度地减少或消除与PD-1阻滞的重叠。我们将使用我们的TRP1 CD8 T细胞自适应转移模型在小鼠中检验这一假设，并使用29 通过PD-1阻断的CDK4/6抑制CDK4/6的I期试验的配对血液和组织样品。我们将使用高最近激活的CD8 T细胞的分辨率单细胞转录分析以跟踪单个TCR clonotypes 及其在CDK4/6抑制剂预处理和治疗血液和组织样品中的转录曲线。影响：对CD4/6在CD8 T细胞命运决策中的机理作用的基本理解将允许更好地设计使用CDK4/6i作为癌症或传染病的临床试验作为调节器，以促进CD8 T细胞记忆。