"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.

项目摘要 /摘要 平均而言，一个男人每16分钟从PCA死亡，这主要是由于继发性恶性肿瘤的发展 原发性癌症部位以外的生长，称为转移。 PCA治疗的基石是 雄激素剥夺疗法（ADT）。 ADT暂时停止PCA，但在几乎所有情况下都会引起抵抗， 导致cast割的PC（CRPC）。然后，CRPC经历了转移性亚克隆的进一步演变 并导致无法治愈的疾病。研究技术揭示了抗性机制和克隆进化 由于当前动物模型模仿PCA进化的能力有限，因此缺乏转移性PCA 天然微环境以及追踪亚克隆进化的不明显方法。 因此，我们开发了回忆（！ 通过使用富含转移性的PTEN/TP53共同消耗来概括人PCA的转移 患者和表观上是通过局灶性开始的原发性疾病发展到骨骼，肺，淋巴结 和肝转移。我们的模型使用慢病毒平台-lv.crebc10携带：（1）CRE（PTEN/TRP53共同 删除； Cas9，荧光和发光标记的激活）； （2）带有十个站点的条形码 由CAS9（BC10）; （3）RNA指南特异性标记BC10; （4）指导或用于测试的短发夹RNA 转移驱动器。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分类。 BC10代表靶向位点的合成阵列，按顺序 降低活性的，对于吸引CAS9产生特定特定编辑的RNA指南。到 流线条形码分析，我们建立了一个R软件包-Evotracer。这个全面的系统 启用：（1）基于原发性和转移中共有的突变模式的癌细胞培养；和 （2）建造系统发育树，以坚固而灵活的方式跟踪转移的进化。 我们的中心假设是，不同的分子和表型克隆结构的差异将是 根据治疗状况，在原发性和转移部位之间精确检测到 转移和/或抗性促进基因和途径。我们的分析将建立和机械 验证由PTEN/TP53-LOSS（基底）引起的转移性克隆扩张的驱动因素，还研究了如何如何研究 在PCA的不同阶段应用的治疗的进化压力（ADT）导致出现 抗性克隆。然后，我们将使用Cas9/Guide（G）RNA和可诱导的短发夹来改变目标基因 那些扩大克隆以识别治疗和治疗诱导的PCA转移的驱动因素。 回忆可以可行地跟踪分子进化并验证药物开发的靶标，这可能导致 新型转移驱动基因和途径的识别。那就是可以在以下方法中应用：（1）主要 疾病，用于早期检测和中断转移的发育； （2）已经存在的转移。 重要的是，该项目中开发的技术也可以应用于其他类型的转移性癌症。