Metastasis of PTEN Mutant Prostate Cancer

PTEN突变前列腺癌的转移

• 批准号: 10540005
• 负责人: Lloyd C Trotman
• 金额: $ 52.94万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10540005
• 关键词: 1-Phosphatidylinositol 3-Kinase; 3-Dimensional; Address; Age; Americas; Animals; Antiandrogen Therapy; Armenia; Autopsy; Back; Body part; Breeding; CRISPR/Cas technology; Candidate Disease Gene; Castration; Cell Culture Techniques; Cell Separation; Cells; Cessation of life; Clinical; Collaborations; Dependence; Development; Diagnosis; Disease; Dreams; Event; Frequencies; Gene Mutation; Generations; Genes; Genetic; Genetically Engineered Mouse; Goals; Histologic; Human; Image; Immunocompetent; In Vitro; Indolent; International; Lentivirus Vector; Life; Malignant Neoplasms; Malignant neoplasm of prostate; Maps; Metastatic Prostate Cancer; Metastatic to; Microscopy; Modeling; Molecular; Molecular Probes; Mus; Mutate; Mutation; Nature; Neoplasm Metastasis; Nude Mice; Oncogenes; Organ; Organoids; PTEN gene; Pathway interactions; Patients; Penetrance; Pharmacology; Phenotype; Phosphotransferases; Positioning Attribute; Problem Solving; Prostate; Proxy; Receptor Protein-Tyrosine Kinases; Recurrent disease; Relapse; Reporting; Research; Resistance; Resolution; Role; Sampling; Signal Transduction; Site; Survivors; System; TP53 gene; Techniques; Testing; Therapeutic; Three-Dimensional Imaging; Time; Transplantation; Validation; Washington; base; candidate validation; cell type; cellular imaging; flexibility; genome analysis; genome sequencing; imaging platform; in vivo; in vivo Model; lead candidate; male; men; mutant; neoplastic cell; programs; prostate cancer model; single cell analysis; standard of care; tomography; tumor; tumor diagnosis; two-photon; upstream kinase; whole genome

With an estimated 190,000 new diagnoses in 2020, Prostate Cancer (PC) is the second most frequent tumor diagnosed in men and the second leading cause of male cancer deaths in America. Approximately one in three men over the age of 50 shows histological evidence of this tumor, however only one in ten will be diagnosed with clinically significant PC. Therapeutic options for localized PC are effective, however, metastatic PC is a yet incurable disease because standard of care anti-androgen therapy invariably results in incurable disease relapse. PTEN-TP53 loss is a most significant event of human lethal metastatic PC as summarized by the PCF/SU2C International Prostate Cancer Dream Team (Armenia et al., 2018) and confirmed by two decades of functional PC modeling in mouse. The central goal of this project is to understand what causes PTEN-mutant indolent prostate cancer (PC) to metastasize to different parts of the body and kill the patient. While the PCF/SU2C report significant association of PTEN loss with loss of TP53, the definition of further genes that are significantly co-mutated with PTEN has remained a problem. Importantly, such genes could point to pathways and principles that drive metastasis and/ or therapy resistance. We aim to solve this problem by combining single cell analysis of patient metastatic rapid autopsy samples with functional genetics and 3D whole organ imaging of lethal metastasis in mouse. We use a highly flexible genetically engineered mouse (GEM) model of lethal, endogenous metastatic prostate cancer, termed RapidCaP. Spontaneous progression to lethal metastasis is seen in ~70% of Pten/Trp53-mutant RapidCaP, suggesting a critical stochastic pioneer event that switches a Pten/ Trp53 null tumor cell from indolence to metastatic escape, similar to a critical pioneering event that causes relapse after castration therapy. However, classical approaches can only capture and reveal the time and physical nature of these pioneering events when already thousands or millions of cells are involved. In Aim 1, we reveal, isolate, and analyze single pioneer cells in space and time. We combine RapidCaP with serial two photon tomography (STPT), which allows us to image whole organs at single cell resolution and construct a 3D map of the time course of escape from indolence and of metastatic relapse after therapy. This guides our isolation of cell types that then serve as validated proxies for pioneer cells of escape or relapse. Through Aim 2, we solve the PTEN co-mutation impasse and define genes co-deleted with PTEN through single cell whole genome analysis of human rapid autopsy samples. We prioritize candidates based on comprehensive functional molecular probing of already isolated pioneer cells in vitro. Through Aim 3, we develop two new modeling platforms for more flexible validation of candidate genes and principles behind escape from indolence and relapse from castration therapy.

2020 年估计有 190,000 例新诊断，前列腺癌 (PC) 是第二常见的肿瘤 诊断为男性，是美国男性癌症死亡的第二大原因。 50 岁以上的男性显示出这种肿瘤的组织学证据，但只有十分之一的人会被诊断出来 对于具有临床意义的 PC，局部 PC 的治疗方案是有效的，但转移性 PC 的治疗方案还很有限。 PCF/SU2C 国际前列腺癌梦团队总结称，标准抗雄激素治疗总是会导致无法治愈的疾病复发，因此 PTEN-TP53 缺失是人类致命性转移性 PC 的最重要事件（亚美尼亚等人，2018 年）。 ）并被二十年的证实 鼠标中的功能性 PC 建模。 该项目的中心目标是了解 PTEN 突变惰性前列腺癌 (PC) 的原因 虽然 PCF/SU2C 报告 PTEN 缺失与 TP53 缺失显着相关，但与 PTEN 显着共突变的其他基因的定义。 重要的是，这些基因可能指出驱动转移的途径和原理。 我们的目标是通过结合患者转移的单细胞分析来解决这个问题。 快速尸检样本，具有功能遗传学和小鼠致死性转移的 3D 全器官成像。 我们使用高度灵活的基因工程小鼠 (GEM) 致命性内源性转移性前列腺模型 癌症，称为 RapidCaP，在约 70% 的 Pten/Trp53 突变型 RapidCaP 中观察到自发进展为致死性转移，这表明存在一个关键的随机先锋事件，将 Pten/Trp53 缺失肿瘤细胞从 转移逃逸的惰性，类似于去势治疗后导致复发的关键开创性事件。然而，经典方法只能捕获并揭示这些开创性事件的时间和物理性质，而这些开创性事件已经涉及数千或数百万个细胞。 在目标 1 中，我们将 RapidCaP 与空间和时间结合起来，揭示、分离和分析单个先锋细胞。 串行双光子断层扫描 (STPT)，它使我们能够以单细胞分辨率对整个器官进行成像 构建摆脱惰性和治疗后转移复发的时间过程的 3D 图。 指导我们分离细胞类型，然后将其作为逃逸或复发先锋细胞的有效代表。 通过目标 2，我们通过对人类快速尸检样本进行单细胞全基因组分析，解决了 PTEN 共突变僵局并定义了与 PTEN 共缺失的基因。目标 3，我们开发两个 新的建模平台可以更灵活地验证候选基因以及摆脱惰性和阉割治疗复发背后的原理。