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: 10693143
• 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/10693143
• 关键词: Adjuvant Chemotherapy; 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; Epithelium; Equipment and Supplies; Evolution; Financial Support; Flow Cytometry; Generations; Genetic Fingerprintings; Genetic Transcription; Genomics; Goals; Growth; Growth Factor; Immune; In Vitro; Institution; Knowledge; Lead; Leadership; Macrophage; 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; System; Systemic Therapy; T-Lymphocyte; Technology; Therapeutic; Time; Tissue Sample; Training; Tumor-infiltrating immune cells; Vision; Work; career; clinical training; combat; crosslink; cytokine; cytotoxicity; experience; gene repair; human tissue; innovation; insight; 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.

项目摘要 高级别浆液性卵巢癌（HGSC）的基因组分析显示，高达50%的HGSC具有 DNA损伤修复（DDR）基因的基因组改变，主要发生在BRCA修复途径中。功能 对患者来源的HGSC类器官的DDR能力的分析显示，超过60%的HGSC具有 停止的复制叉保护DDR路径中的缺陷。这些HGSC类器官的进一步分析 在接受新辅助化疗的患者中，最初具有复制叉的肿瘤 保护缺陷和对卡铂敏感者可诱导接受新辅助化疗 复制叉稳定化并变得对卡铂耐药。新辅助治疗后的转录分析 叉稳定的类器官揭示了这些培养物已经经历了上皮-间充质转化。基于 根据这些结果，假设停滞的复制叉保护DDR路径中的缺陷是 HGSC中的基本分子缺陷，当受到新辅助卡铂治疗的干扰时， 导致分子水平上的缺陷丧失，肿瘤细胞整体状态的改变， 肿瘤细胞与周围基质和免疫细胞相互作用的方式，所有这些联合收割机 创造一个广泛的治疗抗性肿瘤微环境。我所追求的研究挑战是 最初具有叉保护缺陷的肿瘤细胞的进化机制， 失去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 可能会被操纵来防止深度耐药的肿瘤微环境。这部作品将利用一部小说 HGSC类器官共培养系统将是该领域的高度创新，因为它将允许真实的时间 评估DDR缺陷或改变的肿瘤细胞与周围正常细胞之间的相互作用 随着时间和治疗。方法：目标1的目标是为叉的共培养物产生生长条件。 不稳定的铂敏感性类器官，具有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、树突细胞、巨噬细胞和嗜中性粒细胞）。目标2的目标是产生 来自Aim 1的类器官的同基因对是稳定的和不稳定的，然后研究如何相互作用 在类器官共培养系统中，肿瘤细胞与周围基质细胞的比例发生变化， 细胞经历选择性分叉稳定化。目的3的目标是利用不稳定的分叉和 在共培养系统中稳定的来自Aim 2的类器官，并确定肿瘤细胞如何与 周围的免疫细胞随着肿瘤细胞的分叉稳定化而发生变化， 对免疫肿瘤剂的反应。这项工作将对HGSC产生重大影响，因为它将有助于 理解肿瘤-正常细胞相互作用的演变，因为肿瘤细胞叉保护缺陷是 改变和强调。这可能允许开发合理的组合疗法， 靶向肿瘤细胞缺陷，并防止有问题的肿瘤-正常细胞相互作用。

项目成果

Enhanced Efficacy of Aurora Kinase Inhibitors in G2/M Checkpoint Deficient TP53 Mutant Uterine Carcinomas Is Linked to the Summation of LKB1-AKT-p53 Interactions.

• DOI: 10.3390/cancers13092195
• 发表时间: 2021-05-03
• 期刊: Cancers
• 影响因子: 5.2
• 作者: Lynch KN;Liu JF;Kesten N;Chow KH;Shetty A;He R;Afreen MF;Yuan L;Matulonis UA;Growdon WB;Muto MG;Horowitz NS;Feltmate CM;Worley MJ Jr;Berkowitz RS;Crum CP;Rueda BR;Hill SJ
• 通讯作者: Hill SJ

The ORFIUS complex regulates ORC2 localization at replication origins.

• DOI: 10.1093/narcan/zcae003
• 发表时间: 2024-03
• 期刊: NAR cancer
• 影响因子: 5.1