Developing and Automating an Extracellular Vesicle-Based Test for Early Detection of Hepatocellular Carcinoma

开发和自动化基于细胞外囊泡的测试以早期检测肝细胞癌

• 批准号: 10823687
• 负责人: Sean Xiao Liu
• 金额: $ 66.16万
• 依托单位: EXIMIUS DIAGNOSTICS CORP
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-14 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10823687
• 关键词: ANXA5 gene; Ablation; Adopted; Advanced Development; Affect; Alcoholic Liver Diseases; Algorithms; Antibodies; Automation; Bar Codes; Bioinformatics; Biological Markers; Biometry; Blood Circulation; CD81 gene; Cancer Etiology; Case/Control Studies; Cells; Cessation of life; Chemistry; Chronic Hepatitis B; Circulation; Cirrhosis; Clinical; Clinical Practice Guideline; DNA; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Early Diagnosis; Etiology; Excision; Exhibits; GPC3 gene; Goals; Guidelines; Hepatitis B; Hepatitis C; Hepatology; Hour; Individual; Joints; Liver; Liver Cirrhosis; Liver diseases; Mediating; Medical center; Membrane Proteins; Methods; Modality; Motivation; Oncology; Operative Surgical Procedures; Paper; Pathology; Patients; Performance; Phase; Phospholipids; Plasma; Primary Malignant Neoplasm of Liver; Primary carcinoma of the liver cells; Process; Prognosis; Protein Analysis; Quantitative Evaluations; Recommendation; Research; Risk; Robotics; Sampling; Serum; Site; Small Business Innovation Research Grant; Specificity; Specimen; Surface; System; Systems Development; TACSTD1 gene; Testing; Time; Training; Tumor-Derived; Ultrasonography; Validation; Viral hepatitis; alpha-Fetoproteins; arm; assay development; biomarker development; cell type; circulating biomarkers; cohort; detection sensitivity; diagnostic value; early detection biomarkers; extracellular vesicles; hepatocellular carcinoma cell line; in-vitro diagnostics; innovation; lipid nanoparticle; liquid biopsy; liver transplantation; molecular pathology; nanoparticle; neoplastic cell; non-alcoholic fatty liver disease; novel marker; predictive modeling; product development; robotic system; technology platform; tumor; tumor microenvironment; ultrasound

PROJECT SUMMARY Hepatocellular carcinoma (HCC) comprises 80-85% of primary liver cancers and frequently develops in patients with liver cirrhosis or chronic hepatitis B virus infection. HCC's poor prognosis is primarily due to advanced-stage diagnosis. Current clinical practice guidelines recommend biannual liver ultrasounds, with or without serum alpha-fetoprotein (AFP) testing, for at-risk patients to detect HCC at a curable stage. However, their accuracy is limited, with sensitivity between 60-70% and specificity of 90%. Consequently, novel biomarkers for early detection of HCC are urgently needed. Extracellular vesicles (EVs) are a heterogeneous group of lipid nanoparticles that are released by all types of cells, and even more so by tumor cells. Tumor-derived EVs are present in circulation at relatively early stages of disease and are readily accessible across all disease stages. Since the surface proteins of tumor EVs mirror those of the parental tumor cells and those cells within tumor microenvironment, exploiting the diagnostic potential of HCC EVs’ surface protein signatures as a novel biomarker for early detection of HCC holds great promise to significantly augment the ability of current diagnostic modalities. Over the last five years, our joint team comprised of Eximius Dx, UCLA, and Cedars Sinai Medical Center (CSMC) has demonstrated of HCC EV Surface Protein (SP) Test, capable of dissecting and quantifying subpopulations of HCC EVs in plasma samples. In our 2022 Hepatology paper, we summarized a phase-2 biomarker study which successfully validated the feasibility of HCC EV SP Test for early HCC detection. The long-term goal of this Direct-to-Phase-II proposal is to advance the development, optimization, and automation of the HCC EV SP Test, with the ultimate goal of establishing a more sensitive in vitro diagnostic (IVD) test based on HCC EVs. The innovation of the proposed HCC EV SP Test lies in the integration of two platform technologies: (i) EV Click MagBeads for click chemistry-mediated capture of subpopulations of HCC EVs, and (ii) real-time immuno-PCR for quantifying the captured HCC EVs. In parallel, an algorithm will be established to process the resulting HCC EV signatures into HCC EV SP score for distinguishing early-stage HCC from at-risk cirrhosis. This new IVD test will use less then 1-mL plasma and have a sample-to-answer workflow of no more than 3 hours. By adopting an in-house developed robotic system, the automated workflow allows for a throughput > 480 samples per round. Once optimized and automated the HCC EV SP Test will be validated by clinically annotated plasma samples to assess its diagnostic performance for distinguishing early-stage HCC from at-risk liver cirrhotic patients, covering etiologies including alcohol-associated liver disease (ALD), non-alcoholic fatty liver disease (NAFLD), and viral hepatitis (B/C). The successful development of the proposed HCC EV SP Test is rapidly translatable, enabling a sensitive HCC EV-based IVD test for detecting early-stage HCC.

项目总结 肝细胞癌占原发癌的80%-85%，多发于患者。 有肝硬变或慢性乙肝病毒感染。肝细胞癌预后不良的主要原因是晚期 诊断。目前的临床实践指南建议，无论有无血清，一年两次的肝脏超声检查 甲胎蛋白(AFP)检测，用于高危患者在可治愈阶段发现肝细胞癌。然而，他们的精确度是 敏感性为60-70%，特异性为90%。因此，早期的新生物标志物 肝细胞癌的检测迫在眉睫。细胞外小泡(EVS)是一组不同种类的脂质 所有类型的细胞都能释放纳米颗粒，肿瘤细胞释放的纳米颗粒更是如此。肿瘤衍生的电动汽车是 在疾病的相对早期阶段存在于流通中，并且在所有疾病阶段都很容易获得。 由于肿瘤EV的表面蛋白反映了亲代肿瘤细胞和肿瘤内细胞的表面蛋白 微环境，开发肝癌EVS表面蛋白特征的诊断潜力作为一种新的 肝细胞癌早期检测的生物标志物有望显著提高目前的诊断能力 医疗模式。 在过去的五年里，我们的联合团队由Eximius Dx，UCLA和Cedars Sinai医疗中心组成 (CSMC)展示了肝细胞癌EV表面蛋白(SP)检测，能够解剖和量化 血浆样本中肝细胞癌病毒亚群的检测。在我们的2022年肝病论文中，我们总结了第二阶段 生物标志物研究，成功验证了EV SP检测肝癌早期的可行性。这个 此直接到第二阶段计划的长期目标是推进开发、优化和自动化 ，最终目标是建立一种更敏感的体外诊断(IVD)试验 在肝癌电动汽车上。拟议中的HCC EV SP测试的创新之处在于融合了两种平台技术： (I)用于点击化学介导的肝癌EV亚群捕获的EV Click Magbeads，以及(Ii)实时 免疫-聚合酶链式反应定量检测捕获的肝癌EV。同时，将建立一个算法来处理 由此产生的肝细胞癌EV信号纳入肝细胞癌EV SP评分，用于区分早期肝细胞癌和高危肝硬变。 这种新的IVD测试将使用不到1毫升的血浆，样本到答案的工作流程不超过3 几个小时。通过采用内部开发的机器人系统，自动化工作流程允许吞吐量&GT； 每轮480个样品。一旦优化和自动化，肝细胞癌EV SP检测将通过临床验证 用带注释的血浆样本评估其鉴别早期肝癌和高危肝癌的诊断性能 肝硬变患者，包括酒精相关性肝病(ALD)、非酒精性脂肪 肝病(NAFLD)和病毒性肝炎(B/C)。拟议中的肝细胞癌电动汽车SP测试的成功开发 是可快速翻译的，使基于肝细胞癌EV的IVD测试能够灵敏地检测早期肝细胞癌。