Genomic profiling of single circulating tumor cells in the precision medicine of metastatic prostate cancer

项目总结和摘要 转移性去势抵抗性前列腺癌（mCRPC）是前列腺癌（PCa）的最致命状态， 仍然是PCa治疗中最具挑战性的问题。开发更有效的治疗方法， mCRPC是一种显著未满足的临床需求。在治疗反应中存在显著的异质性， mCRPC，需要使用更个性化的策略来指导治疗， 个体患者的基因组特征。使用传统的纸巾有很大的局限性- 基于基因组分析指导mCRPC治疗，部分原因是mCRPC是一种骨为主的转移性肿瘤， 因此，组织样品难以获得，产量通常较低。此外，在治疗过程中， 肿瘤的基因组谱可能会迅速改变以逃避治疗或免疫攻击，导致药物治疗。 阻力为了及时准确地捕捉这些变化并调整治疗方案， 将需要肿瘤活检，这在常规临床实践中难以进行， mCRPC的骨优势性质。因此，为了改善mCRPC预后， 开发新的、非侵入性的液体活检方法，以实时监测治疗反应，并指导 使用不同的治疗方法。循环肿瘤细胞（CTC）从肿瘤脱落到血液中，并且具有非常高的生物学活性。 高恶性潜能，并且可以说是要监测和治疗的肿瘤细胞的最重要的子集。 可以非侵入性地和重复地实时计数，并表现出有希望的预后 潜力，正如FDA批准的用于CTC枚举的CellSearch平台作为 包括mCRPC在内的几种转移性癌症的独立预后因素。然而，国家指导方针 没有一致赞同在常规临床实践中使用CTC计数，主要是因为它 目前尚不清楚对于CTC升高的高危患者应采取何种措施。这些事实凸显了 超越CTC计数和深入CTC基因组表征的重要性。 关于单细胞CTC分析的大规模研究很少报道，部分原因是由于 单CTC检测、分离、全基因组扩增（WGA）和测序偏倚鉴定， 纠正一下我们已经建立了一个关于浓缩、枚举、分离、WGA、 单个CTC的测序和数据分析。基于Sidney Kimmel的三个PCa患者队列 癌症中心，MD安德森癌症中心和乔治华盛顿大学癌症中心，我们将 对单个CTC进行基因组分析，以确定治疗反应和预后的标志物。敬我们最棒 这是mCRPC中第一项基于大规模人群的单CTC分析研究。时发现的问题 本研究将显著提高CTC在mCRPC管理中的临床应用潜力， 精确定制治疗以适应来自个体患者的个体CTC的基因组组成。

PROJECT SUMMARY AND ABSTRACT Metastatic castration-resistant prostate cancer (mCRPC) is the most lethal state of prostate cancer (PCa) and remains as the most challenging issue in PCa treatment. The development of more effective treatments for mCRPC is a significant unmet clinical need. There are substantial heterogeneities in treatment responses of mCRPC, necessitating the use of more personalized strategies in guiding treatment based on the specific genomic characteristics of individual patients. There are considerable limitations in using the traditional tissue- based genomic profiling to guide mCRPC treatment, partly because mCRPC is a bone-predominant metastatic disease, thus tissue samples are difficult to obtain and yields are generally low. Moreover, during treatments, the genomic profiles of tumors may change quickly to evade therapeutic or immune attacks, leading to drug resistance. In order to promptly and accurately capture these changes and adjust treatment plans, repeated tumor biopsies would be needed, which is difficult to perform in routine clinical practices given the invasive and bone-predominant nature of mCRPC. Therefore, in order to improve mCRPC prognosis, it is highly important to develop novel, non-invasive liquid biopsy approaches to real-time monitor treatment response and guide the use of different treatments. Circulating tumor cells (CTCs) are shed from tumors into blood and have extremely high malignant potential, and are arguably the most important subset of tumor cells to monitor and treat. CTCs can be non-invasively and repeatedly enumerated in real-time, and have exhibited promising prognostic potentials, as evidenced by the FDA-approval of the CellSearch platform for CTC enumerations as an independent prognostic factor of several metastatic cancers including mCRPC. However, national guidelines have not unanimously endorsed the use of CTC enumeration in routine clinical practices, mostly because it remains unclear what actions should be taken for high-risk patients with elevated CTCs. These facts highlight the importance of moving beyond CTC enumeration and towards in-depth genomic characterizations of CTCs. Large studies on single-cell CTC analysis have been rarely reported, partly due to the significant challenges on single-CTC detection, isolation, whole genome amplification (WGA), and sequencing bias identification and correction. We have established a comprehensive pipeline on the enrichment, enumeration, isolation, WGA, sequencing, and data analysis of single CTCs. Based on three PCa patient cohorts at the Sidney Kimmel Cancer Center, MD Anderson Cancer Center, and George Washington University Cancer Center, we will conduct genomic profiling of single CTCs to identify markers of treatment response and prognosis. To our best knowledge, this is the first large population-based study of single-CTC analysis in mCRPC. Findings from this study will significantly improve the potential of the clinical application of CTCs in mCRPC management, by precisely tailoring treatment to the genomic make-up of individual CTCs from individual patients.