"Novel Mouse Models for Quantitative Understanding of Baseline and Therapy-Driven Evolution of Prostate Cancer Metastasis"

“用于定量了解前列腺癌转移的基线和治疗驱动演变的新型小鼠模型”

• 批准号: 10660349
• 负责人: DAWID GRZEGORZ NOWAK
• 金额: $ 65.77万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660349
• 关键词: Accounting; Address; Affect; Aftercare; Animal Model; Animals; Architecture; Automobile Driving; Bar Codes; Biological Models; Cancer Prognosis; Cancer Relapse; Castration; Cells; Cessation of life; Clonal Evolution; Clonal Expansion; Computer Models; Computing Methodologies; Consumption; Data; Development; Disease; Disease Progression; Disseminated Malignant Neoplasm; Drug Targeting; Early Diagnosis; Evolution; Fluorescence; Gene Expression Profiling; Genes; Genomics; Goals; Growth; Guide RNA; Heritability; Hormones; Human; Immune system; Interruption; Intervention; Knowledge; Liver; Lung; MYC gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of prostate; Maps; Metastatic Neoplasm to the Liver; Metastatic Prostate Cancer; Methods; Modeling; Molecular; Molecular Evolution; Mutation; Neoplasm Metastasis; Organ; Outcome; PTEN gene; Pathway interactions; Patients; Pattern; Phase; Phenotype; Phylogenetic Analysis; Physiological; Population; Primary Neoplasm; Probability; Process; Prostate; Prostate Cancer therapy; Proto-Oncogenes; Public Health; RB1 gene; RNA; Research; Research Technics; Resistance; Site; Somatic Cell; Sorting; Survival Rate; System; TP53 gene; Technology; Testing; Therapeutic; Time; Tissues; Trees; Tumor Suppressor Proteins; androgen deprivation therapy; animal breeding; bone; cancer cell; cancer site; cancer therapy; curative treatments; design; disorder control; drug development; flexibility; genetic signature; improved; knock-down; luminescence; lymph nodes; man; men; migration; mortality; mouse model; novel; postnatal; pressure; prevent; prostate cancer metastasis; prostate cancer model; resistance mechanism; small hairpin RNA; targeted cancer therapy; tumor; tumor heterogeneity

PROJECT SUMMARY / ABSTRACT On average, a man dies from PCa every 16 minutes, mainly due to development of secondary malignant growths outside of the primary cancer site, known as metastases. The cornerstone of PCa treatment is androgen deprivation therapy (ADT). ADT temporarily halts PCa, but leads to resistance in nearly all cases, resulting in castration-resistant PC (CRPC). CRPC then undergoes further evolution of metastatic subclones and results in incurable disease. Research techniques revealing resistance mechanisms and clonal evolution of metastatic PCa are lacking due to the limited capacity of current animal models to mimic PCa evolution in its native microenvironment as well as inefficient methods for tracing subclonal evolution. Therefore, we developed EvoCaP (!Evolution in Cancer of the Prostate”), a mouse model of endogenous metastasis that recapitulates human PCa genetically, by using PTEN/TP53 co-deletions enriched in metastatic patients, and phenotypically, by focal initiation of primary disease progressing to bones, lungs, lymph nodes and liver metastases. Our model uses a lentiviral platform - LV.CreBC10 carrying: (1) Cre (Pten/Trp53 co- deletions; activation of Cas9, fluorescence and luminescence markers); (2) Barcode with ten sites for marking by Cas9 (BC10); (3) RNA guide specifically marking BC10; and (4) guide or short hairpin RNA for testing metastatic drivers. Luminescence (FLuc) permits continuous tracking of disease progression and fluorescence (eGFP) allows for specific sorting of cancer cells. BC10 represents a synthetic array of on-target sites, in order of decreasing activity, for the RNA guide that attracts Cas9 to generate subsequently specific edits. To streamline barcode analysis, we have established an R package - EvoTraceR. This comprehensive system enables: (1) the profiling of cancer cells based on shared mutational patterns in primary and metastasis; and (2) the building of phylogenetic trees to track evolution toward metastases in a robust and flexible way. Our central hypothesis is that differences in distinct molecular and phenotypical clonal architectures will be precisely detected between primary and metastatic sites depending on therapy status, enabling the inhibition of metastasis and/or resistance promoting genes and pathways. Our analyses will establish and mechanistically validate drivers of metastatic clonal expansion caused by Pten/Tp53-loss (basal) and also investigate how evolutionary pressure from therapy (ADT), applied at different stages of PCa, leads to the emergence of resistant clones. We will then use Cas9/guide (g)RNA and inducible short hairpins to target genes altered in those expanding clones to identify drivers of both treatment-naive and treatment-induced PCa metastasis. EvoCaP can feasibly track molecular evolution and validate targets for drug development, which may lead to identification of novel metastatic driver genes and pathways. Thus, therapies could be applied in: (1) primary diseases for early detection and interruption of metastases development; and (2) already existing metastases. Importantly, technologies developed in this project can also be applied to other types of metastatic cancers.

项目概要/摘要 平均每 16 分钟就有一名男性死于 PCa，主要是由于继发性恶性肿瘤的发展 原发癌症部位以外的生长，称为转移瘤，是 PCa 治疗的基石。 雄激素剥夺疗法 (ADT) 会暂时停止前列腺癌，但几乎在所有病例中都会导致耐药性， 产生去势抵抗性PC（CRPC），然后CRPC进一步进化为转移性亚克隆。 并导致不治之症的研究技术揭示耐药机制和克隆进化。 由于目前的动物模型模拟 PCa 进化的能力有限，因此缺乏对转移性 PCa 的研究。 天然微环境以及追踪亚克隆进化的低效方法。 因此，我们开发了 EvoCaP（！前列腺癌的进化”），一种内源性前列腺癌小鼠模型 通过使用富含转移性的 PTEN/TP53 共缺失，从遗传学上重现人类 PCa 的转移 患者，以及表型上，原发疾病的局灶性起始进展至骨骼、肺、淋巴结 我们的模型使用慢病毒平台 - LV.CreBC10 携带：(1) Cre (Pten/Trp53 co-) 缺失；Cas9、荧光和发光标记的激活）（2）具有十个标记位点的条形码； 由Cas9 (BC10)；(3)特异性标记BC10的RNA向导；和(4)用于测试的向导RNA或短发夹RNA； 发光 (FLuc) 允许连续跟踪疾病进展和荧光。 (eGFP) 允许对癌细胞进行特定排序，BC10 代表按顺序排列的目标位点的合成阵列。 活性降低，因为 RNA 引导吸引 Cas9 生成随后的特定编辑。 简化条形码分析，我们建立了一个 R 包 - EvoTraceR 这个综合系统。 能够：（1）基于原发和转移中共有的突变模式对癌细胞进行分析； （2）建立系统发育树，以稳健而灵活的方式追踪转移的进化。 我们的中心假设是，不同分子和表型克隆结构的差异将是 根据治疗状态精确检测原发部位和转移部位，从而抑制 我们的分析将机械地建立转移和/或耐药性促进基因和途径。 验证 Pten/Tp53 丢失（基础）引起的转移性克隆扩张的驱动因素，并研究如何 治疗的进化压力（ADT）应用于前列腺癌的不同阶段，导致出现 然后，我们将使用 Cas9/guide (g)RNA 和诱导型短发夹来靶向基因。 这些克隆可以识别扩大未经治疗和治疗诱导的 PCa 转移的驱动因素。 EvoCaP 可以切实跟踪分子进化并验证药物开发的目标，这可能会导致 识别新的转移驱动基因和途径因此，疗法可应用于：（1）原发性。 早期发现并阻止转移发展的疾病；(2) 已经存在的转移。 重要的是，该项目开发的技术也可以应用于其他类型的转移性癌症。