Dissecting the interaction between DNA damage repair defects and the tumor microenvironment

剖析DNA损伤修复缺陷与肿瘤微环境之间的相互作用

• 批准号: 10477023
• 负责人: Sarah James Hill
• 金额: $ 44.25万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-16 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10477023
• 关键词: Address; Advisory Committees; Aftercare; B-Lymphocytes; Biological Models; Blood Vessels; Cancer Biology; Cancer Patient; Carboplatin; Cell Communication; Cells; Clinical; Coculture Techniques; Combined Modality Therapy; Complement; DNA; DNA Repair; DNA Sequence Alteration; DNA replication fork; Dana-Farber Cancer Institute; Defect; Dendritic Cells; Development; Disease; Early Diagnosis; Environment; Epithelial; Equipment and Supplies; Evolution; Financial Support; Flow Cytometry; Generations; Genetic Fingerprintings; Genetic Transcription; Genomics; Goals; Growth; Growth Factor; Immune; In Vitro; Knowledge; Lead; Leadership; Malignant neoplasm of ovary; Medical Oncology; Mesenchymal; Molecular; Natural Killer Cells; Neoadjuvant Therapy; Normal Cell; Organoids; Pathway interactions; Patients; Pharmaceutical Preparations; Platinum; Publications; Publishing; Research; Research Personnel; Research Training; Residencies; Resistance; Selection for Treatments; Serous; Stress; Stromal Cells; Stromal Neoplasm; System; Systemic Therapy; T-Lymphocyte; Therapeutic; Time; Tissue Sample; Training; Tumor-Derived; Tumor-infiltrating immune cells; Vision; Work; base; career; chemotherapy; combat; crosslink; cytokine; cytotoxicity; experience; gene repair; human tissue; innovation; insight; macrophage; multidisciplinary; neoplastic cell; neutrophil; novel; prevent; professor; programs; repaired; response; single-cell RNA sequencing; skills; targeted treatment; therapy resistant; treatment response; tumor; tumor microenvironment

PROJECT SUMMARY Genomic analysis of high grade serous ovarian cancer (HGSC) has revealed that up to 50% of HGSCs harbor a genomic alteration in a DNA damage repair (DDR) gene, mostly in the BRCA repair pathway. Functional profiling of DDR capacity of patient derived HGSC organoids has revealed that over 60% of HGSCs harbor defects in the stalled replication fork protection DDR pathway. Further analysis of these HGSC organoids reveals that in patients who receive neoadjuvant chemotherapy, a tumor which initially had a replication fork protection defect and was carboplatin sensitive can be induced by the neoadjuvant chemotherapy to undergo replication fork stabilization and become carboplatin resistant. Transcriptional analysis of the post-neoadjuvant fork stable organoids reveals that these cultures have undergone an epithelial-mesenchymal transition. Based on these results, the hypothesis is that defects in the stalled replication fork protection DDR pathway are a fundamental molecular defect in HGSC that when perturbed by treatment with neoadjuvant carboplatin can lead to loss of the defect at the molecular level, alterations in the overall state of the tumor cells, and changes in the way the tumor cells interact with the surrounding stroma and immune cells, all of which combine to create a broadly therapy resistant tumor microenvironment. The research challenge I will pursue is to dissect the evolving mechanisms by which tumor cells which originally harbor fork protection defects and are induced to lose the defect interact with the surrounding stroma and immune cells and how these changing interactions might be manipulated to prevent a deeply resistant tumor microenvironment. The work will utilize a novel HGSC organoid co-culture system which will be highly innovative for the field because it will allow real time assessment of the interactions between DDR defective or altered tumor cells and the surrounding normal cells over time and treatment. Approach: The goal of Aim 1 is to generate growth conditions for co-cultures of fork unstable platinum sensitive organoids with patient matched intra-tumoral stromal cells and all immune cells (T cells, B cells, NK cells, dendritic cells, macrophages, and neutrophils). The goal of Aim 2 will be to generate isogenic pairs of the organoids from Aim 1 that are fork stable and unstable and then study how the interaction of the tumor cells with the surrounding stromal cells changes in the organoid co-culture system as the tumor cells undergo selective fork stabilization. The goal of Aim 3 is to utilize the isogenic pairs of fork unstable and stable organoids from Aim 2 in the co-culture system and determine how the interaction of the tumor cells with the surrounding immune cells changes as the tumor cells undergo fork stabilization and how this may alter response to immuno-oncologic agents. This work will have major impact in HGSC because it will help understand the evolution of the tumor-normal cell interaction as the tumor cell fork protection defects are altered and stressed. This may allow for the development of rational combination therapies that simultaneously target the tumor cell defects and also prevent problematic tumor-normal cell interactions.

项目总结 高级别浆液性卵巢癌(HGSC)的基因组分析显示，高达50%的HGSC DNA损伤修复(DDR)基因的一种基因组改变，主要发生在BRCA修复途径中。功能性 对患者来源的HGSC有机物的DDR能力的分析显示，超过60%的HGSC 停滞的复制分叉保护DDR途径中的缺陷。HGSC有机化合物的进一步分析 研究表明，在接受新辅助化疗的患者中，一种最初有复制叉子的肿瘤 保护缺陷和卡铂敏感可诱导接受新辅助化疗 复制分叉稳定，并对卡铂产生抗药性。新佐剂后基因的转录分析 分叉稳定的有机体表明，这些培养物经历了上皮向间充质的转变。基座 根据这些结果，假设停滞的复制分叉保护DDR途径中的缺陷是一种 HGSC中的基本分子缺陷，当被新辅助卡铂治疗时，可以 导致分子水平上缺陷的丢失，肿瘤细胞整体状态的改变，以及 在肿瘤细胞与周围基质和免疫细胞相互作用的方式上，所有这些细胞结合在一起 创造一个广泛耐受治疗的肿瘤微环境。我将面临的研究挑战是剖析 最初含有叉状保护缺陷并被诱导的肿瘤细胞的进化机制 丢失缺陷与周围基质和免疫细胞的相互作用以及这些变化的相互作用 可能会被操控，以防止对肿瘤产生深度抗药性的微环境。这部作品将利用一部小说 HGSC有机化合物共培养系统，这将是该领域的高度创新，因为它将允许实时 DDR缺陷或改变的肿瘤细胞与周围正常细胞相互作用的评估 随着时间的推移和治疗。方法：目标1的目标是为叉子的共培养创造生长条件 不稳定的铂敏感类有机物与患者匹配的肿瘤内基质细胞和所有免疫细胞(T 细胞、B细胞、NK细胞、树突状细胞、巨噬细胞和中性粒细胞)。目标2的目标将是产生 来自目标1的稳定和不稳定的有机化合物的同基因对，然后研究如何相互作用 肿瘤细胞与周围基质细胞在类器官共培养体系中的变化 细胞经历选择性分叉稳定化。目标3的目标是利用分叉不稳定和不稳定的等位基因对 在共培养系统中来自Aim 2的稳定的有机物，并确定肿瘤细胞与 当肿瘤细胞经历分叉稳定时，周围的免疫细胞发生变化，以及这种变化可能如何改变。 对免疫肿瘤学药物的反应。这项工作将对HGSC产生重大影响，因为它将有助于 了解肿瘤-正常细胞相互作用的演变，因为肿瘤细胞叉状保护缺陷是 变了，变得紧张。这可能允许开发合理的联合疗法，同时 靶向肿瘤细胞缺陷，并防止有问题的肿瘤与正常细胞的相互作用。