Preclinical analyses of advanced prostate cancer in genetically-engineered mice

项目摘要/摘要 晚期前列腺癌通常是导致男性癌症死亡的主要原因。 治疗失败，这会导致高度转移性的侵袭性疾病变体。在主要的 在晚期前列腺癌中，与DNA修复相关的生物过程是因果性失调而发生的。 值得注意的是，BRCA1和BRCA2是晚期前列腺癌中最常见的DNA修复基因 癌症，也是其治疗的关键靶点。为了研究它们在前列腺癌中的作用，我们产生了 基于BRCA1和BRCA2可诱导功能丧失的基因工程小鼠模型 前列腺。在初步研究中，我们发现BRCA2功能丧失会导致侵袭性前列腺 具有高度穿透性转移的肿瘤，包括骨转移，与DNA损伤增加和 由于缺乏雄激素而加速死亡。同时，我们开发了计算工具来解释 BRCA1和BRCA2的分子决定因素在前列腺癌中的作用。我们还开发了一种新的 名为OncoLoop的精确肿瘤学平台，通过计算将单个患者与单个GEMM进行匹配， 并预测和验证针对特定患者-GEMM对的药物。 利用这些模型和资源，我们将系统地研究BRCA1和BRCA1的功能 BRCA2在前列腺癌中的作用，受DNA修复缺陷在前列腺癌中起重要作用的假设的指导 前列腺癌的进展和治疗，特别是在雄激素缺乏的情况下。在目标1中，我们将 研究在GEMM中BRCA1和BRCA2功能丧失的后果，GEMM衍生的有机物 前列腺癌发生、转移和DNA损伤的模型和患者衍生的人类器官模型 回应。这些研究将为了解BRCA1和BRCA2在前列腺中的功能提供基本的见解 并阐明DNA修复机制与疾病进展的关系。在目标2中，我们将 通过鉴定和功能研究前列腺癌DNA修复功能的分子决定因素 表征代表BRCA1和BRCA2损失的机械性决定因素的主调节器(MRS) 在前列腺癌和转移瘤中的作用。MRS将基于对人类前列腺的保护而优先考虑 癌症，并在小鼠和人类器官中进行了功能验证。我们将通过以下方式补充这些研究 用单核RNA测序(SNRNA-SEQ)分析肿瘤和转移瘤的MR信号 与特定的单元状态相关联。在目标3中，我们将利用我们的GEMM、GEMM衍生的有机化合物和 同种异体移植和患者衍生的器官模型进行DNA修复功能的联合临床研究 前列腺癌。在目标3A中，我们将系统地评估目前正在或正在进行的药物/药物组合 到临床实践。在AIM 3B中，我们将使用OncoLoop平台来预测靶向DNA修复的新药 特定于前列腺的环境。总体而言，这些研究将提供生物学和分子方面的见解以供参考 目前的试验和支持未来试验的发展。

Project Summary/Abstract Advanced prostate cancer represents a major cause of cancer death in men, usually as a consequence of treatment failure, which gives rise to aggressive disease variants that are highly metastatic. Among the major biological processes that are causally dysregulated in advanced prostate cancer are those related to DNA repair. Notably, BRCA1 and BRCA2, are among the most frequently altered DNA repair genes in advanced prostate cancer, and are also key targets for its treatment. To study their functions in prostate cancer, we have generated genetically-engineered mouse models (GEMMs) based on inducible loss-of-function of Brca1 and Brca2 in the prostate. In preliminary studies, we have found that loss-of-function of Brca2 results in aggressive prostate tumors with highly penetrant metastases, including to bone, that are coincident with increased DNA damage and accelerated by androgen deprivation. In parallel, we have developed computational tools to elucidate the molecular determinants of Brca1 and Brca2 functions in prostate cancer. We have also developed a new precision oncology platform called OncoLoop, to computationally match individual patients to individual GEMMs, and to predict and validate drugs that target specific patient-GEMM pairs. Leveraging these models and resources, we will systematically investigate the functions of BRCA1 and BRCA2 in prostate cancer, guided by the hypothesis that defective DNA repair plays an important role in prostate cancer progression and treatment, particularly in the context of androgen deprivation. In Aim 1, we will investigate the consequences of loss-of-function of Brca1 and Brca2 in GEMMs, GEMM-derived organoid models, and patient-derived human organoid models for prostate tumorigenesis, metastasis, and DNA damage response. These studies will provide fundamental insights into the functions of BRCA1 and BRCA2 in prostate cancer and elucidate the relationship of DNA repair mechanisms for disease progression. In Aim 2, we will investigate molecular determinants of DNA repair functions in prostate cancer by identifying and functionally characterizing master regulators (MRs) that represent mechanistic determinants of Brca1 and Brca2 loss-of- function in prostate tumors and metastases. MRs will be prioritized based on conservation with human prostate cancer, and functionally validated in mouse and human organoids. We will complement these studies by analyses of tumors and metastases using single-nuclei RNA-sequencing (snRNA-seq) to identify MR signatures associated with specific cell states. In Aim 3, we will leverage our GEMMs, GEMM-derived organoids and allografts, and patient-derived organotypic models to pursue co-clinical investigations of DNA repair function in prostate cancer. In Aim 3A, we will systematically evaluate drugs/drug combinations currently in or advancing to clinical practice. In Aim 3B, we will use the OncoLoop platform to predict new drugs that target DNA repair in prostate-specific contexts. Overall, these studies will provide biological and molecular insights to inform on current trials and to support the development of future ones.