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

项目摘要和摘要 转移性去势抵抗前列腺癌(MCRPC)是前列腺癌(PCA)最致命的状态， 仍然是PCa治疗中最具挑战性的问题。更有效的治疗方法的发展 MCRPC是一个重要的未得到满足的临床需求。在治疗反应上有很大的异质性。 MCRPC，有必要根据具体情况使用更个性化的策略来指导治疗 个体患者的基因组特征。使用传统的纸巾有相当大的局限性- 基于基因组图谱来指导mCRPC治疗，部分原因是mCRPC是一种以骨为主的转移瘤 疾病，因此组织样本很难获得，产量普遍较低。此外，在治疗过程中， 肿瘤的基因组图谱可能会迅速改变，以逃避治疗或免疫攻击，从而导致药物 抵抗。为了及时准确地捕捉到这些变化，调整治疗方案，反复 将需要肿瘤活检，这在常规临床实践中很难进行，因为具有侵袭性和 MCRPC的骨主性。因此，为了改善mCRPC的预后，至关重要。 开发新的非侵入性液体活组织检查方法，实时监测治疗反应并指导 使用不同的治疗方法。循环中的肿瘤细胞(CTCs)从肿瘤进入血液，并具有极高的 恶性潜能高，可以说是需要监测和治疗的最重要的肿瘤细胞亚群。CTCS 可以是非侵入性的，并实时重复计数，并显示出良好的预后 潜力，正如FDA批准的CellSearch平台的CTC枚举为 包括mCRPC在内的几种转移癌的独立预后因素。然而，国家指导方针 没有一致赞成在常规临床实践中使用CTC枚举法，主要是因为它 目前尚不清楚对于CTCs升高的高危患者应该采取什么措施。这些事实突显了 必须超越四氯化碳的列举，深入研究四氯化碳的基因组特征。 关于单细胞CTC分析的大型研究很少报道，部分原因是 单CTC检测、分离、全基因组扩增(WGA)和测序偏倚鉴定 更正。我们已经建立了关于浓缩、枚举、分离、WGA、 对单个CTCs进行测序和数据分析。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进行基因组图谱分析，以确定治疗反应和预后的标志物。为我们最好的 知识，这是mCRPC中第一个基于大规模人群的单一四氯化碳分析研究。由此得出的结论 这项研究将显著提高CTCS在mCRPC管理中的临床应用潜力，通过 针对单个患者的单个CTC的基因组组成进行精确的定制治疗。

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.