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.

项目摘要 肝细胞癌（HCC）占原发性肝癌的80 - 85%，并且经常在患者中发生 肝硬化或慢性B型肝炎病毒感染者。肝癌的预后差主要是由于晚期 诊断.目前的临床实践指南推荐一年两次的肝脏超声检查，有或没有血清 甲胎蛋白（AFP）检测，用于高危患者在可治愈阶段检测HCC。然而，其准确性 有限，灵敏度为60 - 70%，特异性为90%。因此，新的生物标志物的早期 因此，迫切需要检测HCC。细胞外囊泡（EV）是一组异质的脂质 所有类型的细胞都释放出纳米颗粒，肿瘤细胞更是如此。肿瘤衍生的EV是 在疾病的相对早期阶段存在于循环中，并且在所有疾病阶段都容易获得。 由于肿瘤EV的表面蛋白反映了亲本肿瘤细胞和肿瘤内的那些细胞的表面蛋白， 微环境，利用HCC EV的表面蛋白特征作为新的诊断潜力， 早期检测HCC的生物标志物有很大的希望，以显着提高目前的诊断能力， 方式。 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）已证明HCC EV表面蛋白（SP）检测，能够解剖和定量 血浆样品中HCC EV的亚群。在我们2022年的肝病学论文中，我们总结了一个2期 生物标志物研究成功验证了HCC EV SP测试用于早期HCC检测的可行性。的 这个直接进入第二阶段的提案的长期目标是推进开发、优化和自动化 HCC EV SP检测的最终目标是建立一种更灵敏的体外诊断（IVD）检测， 在HCC EV上。建议的HCC EV SP测试的创新在于两种平台技术的集成： (i)EV Click MagBeads用于HCC EV亚群的点击化学介导的捕获，和（ii）实时 免疫PCR用于定量捕获的HCC EV。并行地，将建立算法来处理 将所得到的HCC EV特征转化为HCC EV SP评分，用于区分早期HCC与高危肝硬化。 这种新的IVD检测将使用不到1 mL的血浆，样本到答案的工作流程不超过3个 小时通过采用内部开发的机器人系统，自动化的工作流程可以实现> 每轮480个样本。一旦优化和自动化，HCC EV SP检测将通过临床验证 注释的血浆样本，以评估其区分早期HCC与风险HCC的诊断性能 肝病患者，涵盖病因包括酒精相关性肝病（ALD）、非酒精性脂肪性肝病、 肝病（NAFLD）和病毒性肝炎（B/C）。成功开发拟议的HCC EV SP测试 可快速转换，从而实现用于检测早期HCC的灵敏的基于HCC EV的IVD测试。