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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要肝细胞癌 (HCC) 是全球癌症相关死亡的第二大常见原因。肝移植（LT）是对于患有不可切除、非转移性疾病的 HCC 患者的唯一治疗方法符合严格的放射学肿瘤大小和数量标准（米兰标准）。符合资格的候选人将经历有限的观察期（>6 个月）可用于评估肿瘤生物学，但本质上存在 HCC 风险 1 年后，15-20% 的患者出现病情进展和等待名单退出。尽管有这些选择实践，LT后 HCC 复发困扰着高达 20% 的患者，是同种异体移植失败和患者死亡的主要原因。那里迫切需要能够动态监测肿瘤生物学的非侵入性生物标志物，以更好地平衡等待名单中退出和 LT 后复发的风险，并允许改进 HCC 患者接受治疗的优先顺序同种异体肝移植稀缺。该提案旨在开发一种基于血液的综合分析（即 NanoVelcro vimCTC 检测用于检测波形蛋白 + 循环肿瘤细胞 [CTC] 和 HCC CTC-RNA 检测用于 HCC- 特定 RNA 特征）用于等待名单上的 HCC 患者，以确定最适合 LT 的患者。集成的该测定将为研究 HCC CTC 的表型和分子特征提供一种新方法。使用 HCC 特异性多标记物捕获混合物和优化的免疫细胞化学 (ICC) 染色协议中，提议的 NanoVelcro vimCTC 检测能够识别 HCC CTC 的亚群波形蛋白表达（称为 vimCTC、DAPI+/CK+/CD45-/波形蛋白+）。该亚群与接受治疗性治疗的临床上无法区分的早期患者的复发率增加治疗。 HCC CTC-RNA 检测是将 CTC 分离与 Click Chip 相结合而开发的，具有点击化学介导的细胞捕获和二硫键裂解细胞释放，以及下游 RNA 表达使用 NanoString 的 nCounter 平台对纯化的 CTC 进行分析。这允许准确量化以非侵入性方式分析 HCC CTC 衍生的 mRNA 标记物。由此产生的 vimCTC 计数和 mRNA 配置文件有望增强当前 LT 候选选择算法的能力。拟议的研究将通过具体目标 1a 实施：在银行进行回顾性研究使用 NanoVelcro vimCTC 检测的血液样本可细化 vimCTC 计数与术后处理之间的关联 LT 复发/等待名单退出。；具体目标 1b：对新鲜采集的血液进行前瞻性研究用于确定 vimCTC 计数与 LT 后复发/等待名单退出之间关联的样本；和具体目标 2：对新鲜采集的血液样本进行前瞻性研究以确定相关性 HCC CTC-RNA 检测与 LT 后复发/等待名单退出之间的关系。评估的中心假设将 vimCTC 和侵袭性 RNA 特征的基线和纵向变化将显着改善当前临床放射学 LT 选择标准预测 LT 后复发和候补名单退出的能力，为基于肿瘤生物学的 HCC LT 候选者选择实践铺平道路。