Microfluidic isolation and molecular analysis of circulating tumor cells for the study of neuroendocrine transdifferentiation in prostate cancer

项目摘要/摘要 尽管下一代雄激素受体(AR)靶向治疗提高了患者的存活率 前列腺癌患者，最近的研究表明，接触这些治疗方法会导致转分化 近五分之一的病例表现为侵袭性神经内分泌(NE)表型。分子决定因素 前列腺癌的谱系可塑性还知之甚少，而且对非侵袭性的需求还没有得到满足。 方法研究治疗诱导的谱系可塑性随时间的变化。我们的团队在 马萨诸塞州总医院(MGH)最近开发了一种新型微流控芯片LPCTC-iChip，用于 从整个白细胞分离产品中高通量分离循环肿瘤细胞(CTC)，因此 为实时分子图谱的连续肿瘤活检提供了一种非侵入性的替代方法。在这 校外校内协作以患者为中心的翻译研究项目，我们建议 将LPCTC-iChip纳入NCI的临床试验以研究NE的转分化 前列腺癌。该应用程序的总体目标是洞察时间发展， 分子决定因素，通过分析CTCs与非CTCs比较，NE转分化的临床影响 美国国立卫生研究院临床化学免疫治疗试验中前列腺癌患者的匹配肿瘤活检组织 中心。我们的中心假设是CTC分析可以捕捉到前列腺的动态谱系状态 治疗期间和治疗后的癌症，免疫反应可以延缓NE的临床影响 表型。为了实现总体目标，将追求以下三个具体目标：1)评估 匹配的CTC和转移性去势患者的转移性活检组织- 耐药前列腺癌的NE表型特征：2)评估慢性前列腺癌患者的系列CTC 转移性去势敏感型前列腺癌治疗期间去甲肾上腺素转分化；3)确定 免疫反应、NE表型和临床结果之间的关系。LPCTC-iChip将 实际放置在NIH临床中心，以及来自mCRPC和MCRPC的连续CTC和匹配的活检 M7824+M9241+多西紫杉醇化疗免疫治疗mCSPC的研究 (NCT04633252)将分析NE表型的基因组、转录和蛋白质标记。 这项拟议的研究具有创新性，因为它利用了一种新的微流控设备LPCTC-iChip，用于 分离CTC以询问前列腺癌中一个高度相关的问题。这项研究具有重要意义 因为它有望提供一种非侵入性的替代组织活检来监测治疗- 诱导的谱系可塑性，以及为未来临床设计提供了强有力的科学框架 预防和治疗致死性NE表型前列腺癌的试验。这种校内校外的合作 利用来自MGH和NCI的独特资源来了解重要的疾病过程-NE 转分化，对前列腺癌治疗监测和治疗的影响。

PROJECT SUMMARY / ABSTRACT Although next-generation androgen receptor (AR)-targeted therapies have increased survival in prostate cancer patients, recent studies reveal that exposure to these therapies result in transdifferentiation to an aggressive neuroendocrine (NE) phenotype in nearly one fifth of cases. The molecular determinants of lineage plasticity in prostate cancer are poorly understood, and there is an unmet need for non-invasive methods to study the emergence of treatment-induced lineage plasticity over time. Our team at the Massachusetts General Hospital (MGH) recently developed a novel microfluidic chip, the LPCTC-iChip, for the high-throughput isolation of circulating tumor cells (CTCs) from an entire leukapheresis product, thus providing a non-invasive alternative to serial tumor biopsies for real-time molecular profiling. In this extramural-intramural collaborative patient-centric translational research project, we propose to incorporate the LPCTC-iChip into an NCI intramural clinical trials to investigate NE transdifferentiation in prostate cancer. The overall objective of this application is to gain insight into the temporal development, molecular determinants, and clinical impact of NE transdifferentiation by analyzing CTCs in comparison to matched tumor biopsies from prostate cancer patients in a chemoimmunotherapy trial at the NIH Clinical Center. Our central hypothesis is that CTC analyses can capture the dynamic lineage state of prostate cancers during and after therapy, and that immune responses can delay the clinical impact of the NE phenotype. To attain the overall objective, the following three specific aims will be pursued: 1) Evaluate matched CTCs and metastatic biopsies from patients with heavily pre-treated metastatic castration- resistant prostate cancer for hallmarks of NE phenotype; 2) Evaluate serial CTCs in patients with metastatic castration-sensitive prostate cancer for NE transdifferentiation during therapy; and 3) Determine the association between immune responses, NE phenotype, and clinical outcomes. The LPCTC-iChip will be physically placed at the NIH Clinical Center, and serial CTCs and matched biopsies from mCRPC and mCSPC patients enrolled in the chemoimmunotherapy study of M7824+M9241+docetaxel (NCT04633252) will be analyzed for genomic, transcriptional, and protein markers of the NE phenotype. The proposed research is innovative because it utilizes a novel microfluidic device, the LPCTC-iChip, for the isolation of CTCs to interrogate a highly relevant problem in prostate cancer. The research is significant because it is expected to provide a non-invasive alternative to tissue biopsy for monitoring treatment- induced lineage plasticity, as well as provide a strong scientific framework for the design of future clinical trials to prevent and treat lethal NE phenotype prostate cancers. This intramural-extramural collaboration leverages unique resources from MGH and NCI to understand an important disease process, NE transdifferentiation, with implications for prostate cancer treatment monitoring and therapy.