DNA damage response and cancer immunity

DNA损伤反应和癌症免疫

基本信息

批准号：
10523886
负责人：
CHRISTOPHER J. BAKKENIST
金额：
$ 46.19万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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：
2022-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10523886
关键词：
Acceleration Adenosine Base Excision Repairs CD8-Positive T-Lymphocytes CD8B1 gene CDC2 gene CT26 Cancer Model Cancer Patient Cell Cycle Cell Cycle Checkpoint Cell Cycle Kinetics Cells Cessation of life Chemotherapy and/or radiation Clinical Trials Cyclin-Dependent Kinases Cytidine DNA DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway DNA Sequence DNA biosynthesis DNA replication fork DNA-Directed RNA Polymerase Data Dose-Limiting Excision Excision Repair Genetic Transcription Genetically Engineered Mouse Genome Genome Stability Genomic DNA Goals Guanosine Immune Immune response Immunologic Memory Immunotherapy In Vitro Inbred BALB C Mice Interferons Ionizing radiation Kinetics Knock-out Laboratories Licensing Lymphopenia MCM4 gene Malignant Neoplasms Mediating Mus Mutation Phosphotransferases RNA Radiation therapy Reporting Ribonucleosides Ribonucleotides S phase Signal Transduction System T cell response T cell therapy Testing Thymidine Toxic effect Transplantation Tumor Immunity Tumor-infiltrating immune cells Uracil Uridine anti-tumor immune response ataxia telangiectasia mutated protein cancer cell cell killing cell type chemotherapy helicase immune checkpoint blockade in vivo inhibitor innovation kinase inhibitor mouse model novel strategies repaired response transplant model tumor uracil-DNA glycosylase

项目摘要

The overarching goal of our laboratory is to determine how DNA damage response inhibitors (DDRi) can be used to potentiate cancer cell killing while concurrently increasing anti-tumor immune responses after radiation therapy (XRT). The DNA Damage Response (DDR) is a signaling system that integrates DNA repair pathways and the cell cycle to safeguard genome stability. In addition to activating cell cycle checkpoints and DNA repair in cells treated with XRT, the DDR limits origin firing and delays cell cycle transitions in unstressed cells. While cyclin- dependent kinases are cell cycle accelerators, DDR kinases are cell cycle brakes and, in this analogy, DDRi disable the brakes, causing unchecked acceleration. Here we will determine how the DDR is rewired in CD8+ T cells to accommodate massive and concomitant DNA replication and transcription in S phase. We will also determine the impact of DDRi in cancer and immune cells. We hypothesize that ATR kinase inhibitors induce origin firing that causes ribonucleosides to be mis-incorporated into the genome, and that this generates chimeric RNA-DNA fragments and type I IFN-dependent immunologic memory after XRT. To test our hypothesis in cancer and immune cells, we have generated an innovative transplantable model of cancer. The Mcm4Chaos3/Chaos3 mouse carries a mutation in Mcm4 that destabilizes the replicative helicase. Cells derived from Mcm4Chaos3/Chaos3 mice have a 60% reduction in origin licensing. We have generated Mcm4Chaos3/Chaos3 B16 cancer cells that can be transplanted into Mcm4wt/wt and Mcm4Chaos3/Chaos3 mice. This will allow us to separate the function of ATR that limits origin firing from that which mediates the repair of replication forks in cancer and immune cells. In Aim 1, we will define cell cycle kinetics and determine how ATR inhibitors induce DNA damage in immune and cancer cells in vitro. In Aim 2, we will define cell cycle kinetics and determine whether ATR inhibitors induce DNA damage in immune cells and type 1 interferons in vivo. In Aim 3, we will determine whether ATR inhibitors combine with XRT to generate durable responses and immunologic memory through effects on immune and/or cancer cells. Successful completion of this project will define how the DDR is rewired in CD8+ T cells to accelerate cell cycle transitions and accommodate massive and concomitant DNA replication and transcription in S phase which, accounts for ~70% of the cell cycle as G1 is abridged. These studies are highly significant as the objective of checkpoint blockade and adoptive T cell transfer is to induce rapid division in CD8+ T cells. Successful completion of this project will identify combinations and sequences of DDRi that potentiate cancer cell killing while concurrently increasing anti-tumor immune responses in mouse models of cancer treated with XRT. These studies are highly significant as we use DDRi that are currently in 115 clinical trials and XRT which is used to treat >50% of cancer patients, >60% with curative intent.

我们实验室的首要目标是确定如何使用 DNA 损伤反应抑制剂 (DDRi) 增强癌细胞杀伤力，同时增强放射治疗后的抗肿瘤免疫反应（XRT）。 DNA 损伤反应 (DDR) 是一个信号系统，整合了 DNA 修复途径和细胞周期以维护基因组稳定性。除了激活细胞周期检查点和细胞内 DNA 修复经过 XRT 处理后，DDR 限制了无应激细胞的起源放电并延迟了细胞周期转变。当细胞周期蛋白- 依赖性激酶是细胞周期加速器，DDR 激酶是细胞周期制动器，在这个类比中，DDRi 禁用刹车，导致不受控制的加速。这里我们将确定CD8+ T中DDR如何重新布线细胞在 S 期适应大量且伴随的 DNA 复制和转录。我们也会确定 DDRi 对癌症和免疫细胞的影响。我们假设 ATR 激酶抑制剂诱导起源激发导致核糖核苷被错误地掺入基因组中，从而产生嵌合体 XRT 后的 RNA-DNA 片段和 I 型 IFN 依赖性免疫记忆。检验我们的癌症假设和免疫细胞，我们已经产生了一种创新的可移植癌症模型。 Mcm4Chaos3/Chaos3 小鼠携带 Mcm4 突变，导致复制解旋酶不稳定。源自 Mcm4Chaos3/Chaos3 的细胞小鼠的原产地许可减少了 60%。我们已经生成了 Mcm4Chaos3/Chaos3 B16 癌细胞，可以移植到 Mcm4wt/wt 和 Mcm4Chaos3/Chaos3 小鼠中。这将使我们能够分离 ATR 的功能限制介导癌症和免疫细胞中复制叉修复的起源发射。在目标 1 中，我们将定义细胞周期动力学并确定 ATR 抑制剂如何在免疫和癌症中诱导 DNA 损伤体外细胞。在目标 2 中，我们将定义细胞周期动力学并确定 ATR 抑制剂是否诱导 DNA 体内免疫细胞和 1 型干扰素的损伤。在目标 3 中，我们将确定 ATR 抑制剂是否与 XRT 结合，通过对免疫和/或癌细胞。该项目的成功完成将定义 DDR 如何在 CD8+ T 细胞中重新连接以加速细胞周期转变并适应大规模且伴随的 DNA 复制和转录处于 S 期，由于 G1 期被缩短，因此约占细胞周期的 70%。这些研究非常重要，因为检查点阻断和过继性 T 细胞转移的目的是诱导 CD8+ T 细胞快速分裂。该项目的成功完成将确定增强癌症的 DDRi 组合和序列细胞杀伤，同时增加抗肿瘤免疫反应的小鼠癌症模型 XRT。这些研究非常重要，因为我们使用目前正在进行 115 项临床试验的 DDRi 和 XRT 用于治疗>50%的癌症患者，>60%具有治愈目的。