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：
10592281
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/10592281
关键词：
ATAC-seq Achievement Adjuvant Adoptive Transfer Affect Affinity Antigens Binding Binding Sites Blood Blood Cells 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 ChIP-seq Chromatin Circulation 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 Immunoprecipitation Immunotherapeutic agent In Vitro Individual Link Malignant Neoplasms Mass Spectrum Analysis Memory Minority Modeling Molecular Mus Nature Nuclear PD-1 blockade Paper Patients Pharmaceutical Preparations Phosphorylation Physiological Positioning Attribute Proteins RNA RUNX3 gene Reporting Resolution Role Sampling Seminal T cell therapy T memory cell T-Cell Activation 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.

项目概要癌症免疫疗法的标志性成就是通过长期持久的方式延长总体生存期然而，记忆 CD8 T 细胞在癌症患者中如何形成的问题仍然存在。我们的长期目标是了解细胞周期调控之间的关系。我们检查了小鼠和人类 CD8 T 细胞在 T 细胞过程中的 CDK4/6 抑制。启动并发现 CD8 T 细胞显着偏向于记忆命运。离体 CDK4/6i 在过继转移至小鼠体内后显示出增强的长寿命记忆反应。进一步使用单细胞转录谱来评估来自血液的最近激活的人类 CD8 T 细胞接受 CDK4/6i palbociclib 或 TCR 克隆型治疗的乳腺癌患者。追踪和 RNA 速度分析表明，CDK4/6 抑制增加了中枢记忆与中枢记忆的比率。配对样本的分析表明，MYC 靶标被下调。 CDK4/6 抑制剂治疗人类，与转录抑制因子 MXD4 的增加一致。中心假设是，小鼠和人类 CD8 T 细胞中的 CDK4/6 抑制可增强长期效果为了测试这一点，我们将使用最先进的技术，包括：1) TRP1 跨核小鼠。由我们实验室开发，用于研究与内源性抗原具有生理亲和力范围的抗原特异性 TCR 2) scRNAseq 和 TCR 克隆型追踪，分析大量纵向血液并进行匹配 II 期试验患者的组织样本；3) ChIPseq 和免疫共沉淀以确定将 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目标 1 中，我们将确定将 CDK4 和/或 CDK6 与诱导记忆细胞命运相关的分子机制。其他细胞周期抑制剂或利用细胞分裂率的自然异质性不会增强 CD8 T 细胞中记忆的形成，意味着 CDK4 和/或 CDK6 在赋予记忆细胞命运方面具有独特的作用。我们将确定受 CDK4/6 抑制影响的转录调节因子，重点关注 MYC 和 RUNX3。 CDK4/6 抑制剂与 PD-1 阻断剂在临床上的组合会导致意想不到的治疗限制在目标 2 中，我们提出短期的 CDK4/6 抑制将成功增强 CD8 T 细胞。记忆，从而允许停止 CDK4/6 抑制剂以最小化或消除与 PD-1 阻断的重叠。将使用我们的 TRP1 CD8 T 细胞过继转移模型在小鼠中测试这一假设，并使用 29 我们将使用来自 CDK4/6 抑制与 PD-1 阻断的 I 期试验的配对血液和组织样本。对最近激活的 CD8 T 细胞进行分辨率单细胞转录分析，以追踪单个 TCR 克隆型及其在 CDK4/6 抑制剂治疗前和治疗中的血液和组织样本中的转录谱。影响：A。对 CD4/6 在 CD8 T 细胞命运决定中的机制作用的基本了解将有助于更好地设计使用 CDK4/6i 作为佐剂促进 CD8 T 细胞记忆的癌症或传染病临床试验。