Integrated analysis of HCC CTCs for Liver Transplant Candidate Selection

用于肝移植候选者选择的 HCC CTC 综合分析

• 批准号: 10117212
• 负责人: Vatche Agopian
• 金额: $ 57.81万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10117212
• 关键词: Algorithms; Allografting; Behavior; Bioinformatics; Biological Assay; Biological Markers; Biology; Biometry; Biopsy; Blood; Blood Banks; Blood specimen; Cancer Etiology; Cell Separation; Cells; Cessation of life; Characteristics; Chemistry; Clinical; Clinical Research; Cryopreservation; Development; Discrimination; Disease; Disulfides; Dropout; Enrollment; Equilibrium; Evaluation; Expression Profiling; Gene Expression; Genes; Genetic Transcription; Goals; Institution; Joints; Liver; Mediating; Messenger RNA; Microfabrication; Microfluidics; Modeling; Molecular; Molecular Profiling; Monitor; Names; NanoVelcro; Nanotechnology; Neoplasm Circulating Cells; Nonmetastatic; Operative Surgical Procedures; PTPRC gene; Pathologic; Pathology; Patient Selection; Patients; Phenotype; Primary carcinoma of the liver cells; Prospective Studies; Protocols documentation; RNA; RNA marker; Radiology Specialty; Recurrence; Research; Retrospective Studies; Risk; Sampling; Selection Criteria; Sensitivity and Specificity; Stains; Stratification; Testing; Time; Tumor Biology; Tumor Markers; Unresectable; Vimentin; Waiting Lists; base; biobank; candidate selection; curative treatments; follow-up; high risk; immunocytochemistry; improved; in-vitro diagnostics; innovation; liver allograft; liver transplantation; mortality; nano; nano-string; nanomaterials; noninvasive diagnosis; novel; novel strategies; primary endpoint; prospective; standard of care; transcriptomics; tumor; tumor progression

PROJECT SUMMARY Hepatocellular carcinoma (HCC) is the 2nd most common cause of cancer-related deaths worldwide. Liver transplantation (LT) is the only curative therapy for HCC patients with unresectable, non-metastatic disease who meet strict radiologic tumor size and number criteria (Milan criteria). Eligible candidates undergo a finite observation period (>6 months) that allows for an assessment of tumor biology, but which inherently risks HCC progression and wait-list dropout in 15-20% of patients after 1 year. Despite these selection practices, post-LT HCC recurrence plagues up to 20% of patients, and is a major cause of allograft loss and patient mortality. There is a dire need for non-invasive biomarkers capable of dynamic monitoring of tumor biology to better balance the risk of wait-list dropout and post-LT recurrence, and allowing for improved prioritization of HCC patients to receive scarce liver allografts. This proposal aims to develop an integrated blood-based analysis (i.e., NanoVelcro vimCTC Assay for detecting vimentin+ circulating tumor cells [CTCs] and HCC CTC-RNA Assay for HCC- specific RNA signatures) for wait-listed HCC patients, to identify patients most suitable for LT. The integrated assay will provide a novel approach to study both phenotypic and molecular characteristics of HCC CTCs. Using an HCC-specific multi-marker capture cocktail and optimized immunocytochemistry (ICC) staining protocol, the proposed NanoVelcro vimCTC Assay is capable of identifying a subpopulation of HCC CTCs with vimentin expression (named vimCTC, DAPI+/CK+/CD45-/vimentin+). This subpopulation is associated with increased recurrence in the subset of clinically indistinguishable early-stage patients undergoing curative-intent treatment. The HCC CTC-RNA Assay was developed by combining CTC isolation with Click Chip, featuring click chemistry-mediated cell capture and disulfide-cleavage cell release, with downstream RNA expression profiling of the purified CTCs with NanoString's nCounter platform. This allows for accurate quantification of a panel of HCC CTC-derived mRNA markers in a non-invasive manner. The resulting vimCTC counts and mRNA profiles hold great promise to augment the ability of the current LT candidate selection algorithm. The proposed research will be implemented via Specific Aim 1a: Conducting a retrospective study in banked blood samples using NanoVelcro vimCTC Assay to refine the association between vimCTC counts and post- LT recurrence/wait-list dropout.; Specific Aim 1b: Conducting a prospective study on freshly collected blood samples to determine association between vimCTC counts and post-LT recurrence/wait-list dropout; and Specific Aim 2: Conducting a prospective study on freshly collected blood samples to determine the association between HCC CTC-RNA Assay and post-LT recurrence/wait-list dropout. The central hypothesis evaluated will be that baseline and longitudinal changes in vimCTCs and aggressive RNA signatures will significantly improve the ability of current clinicoradiologic LT selection criteria in predicting post-LT recurrence and wait-list dropout, paving the way for tumor-biology based HCC LT candidate selection practices.

项目摘要 肝细胞癌（HCC）是全球与癌症相关死亡的第二大原因。肝 移植（LT）是唯一的治疗疗法，用于患有不可切除的非转移性疾病的HCC患者 符合严格的放射学肿瘤大小和数字标准（米兰标准）。合格的候选人会经历有限的 观察期（> 6个月）可以评估肿瘤生物学，但固有的风险HCC风险 1年后，15-20％的患者的进展和等待名单辍学。尽管有这些选择实践，但 HCC复发困扰着多达20％的患者，这是同种异体移植丧失和患者死亡率的主要原因。那里 是否需要动态监测肿瘤生物学的非侵入性生物标志物，以更好地平衡 等待列表辍学和LT复发的风险，并允许改善HCC患者的优先次序 稀缺的肝脏同种异体移植物。该提案旨在开发基于血液的综合分析（即纳米诺维尔科 用于检测波形蛋白+循环肿瘤细胞[CTC]和HCC CTC-RNA分析的VIMCTC测定法 特定的RNA签名）等待上市的HCC患者，以识别最适合LT的患者。集成 测定将提供一种研究HCC CTC的表型和分子特征的新方法。 使用HCC特定的多标记捕获鸡尾酒和优化的免疫细胞化学（ICC）染色 协议，提出的纳米诺维克罗vimctc分析能够识别使用HCC CTC的亚群 波形蛋白表达（命名为vimctc，dapi+/ck+/cd45-/vimentin+）。该亚群与 在接受治愈性的早期患者的子集中的复发增加 治疗。 HCC CTC-RNA分析是通过将CTC隔离与点击芯片相结合的，以 单击化学介导的细胞捕获和二硫化裂解细胞的释放，下游RNA表达 使用NanoString的NCounter平台对纯化的CTC进行分析。这允许准确量化 HCC CTC衍生的mRNA标记的面板以非侵入性方式。由此产生的VIMCTC计数和mRNA 配置文件具有巨大的希望，可以增强当前LT候选算法的能力。 拟议的研究将通过特定的目标1A实施：在银行中进行回顾性研究 使用Nanovelcro vimctc测定的血液样本来完善VIMCTC计数与后的关联 lt recurrence/watch-list辍学。特定目标1B：对新鲜收集的血液进行前瞻性研究 样本以确定VIMCTC计数与LT复发/等待名单辍学之间的关联；和 特定目的2：对新鲜收集的血液样本进行前瞻性研究以确定关联 在HCC CTC-RNA分析和LT复发/等待名单辍学之间。评估的中央假设将 VIMCTC的基线和纵向变化和攻击性RNA特征将显着改善 当前临床原理LT选择标准在预测LT复发和等待名单辍学的能力， 为基于肿瘤生物的HCC LT候选选择实践铺平道路。