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Principal Investigator: Vatche Agopian
Organization: UNIVERSITY OF CALIFORNIA LOS ANGELES
Fiscal Year: 2024
Award: $628,431
Funding agency: National Cancer Institute
PROJECT SUMMARY
Hepatocellular carcinoma (HCC) accounts for 80-85% of primary liver cancers, and mainly occurs in patients
with liver cirrhosis or chronic hepatitis B virus (HBV) infection. Prognosis of HCC is dismal primarily due to
advanced stage of disease at diagnosis. Current clinical practice guidelines recommend HCC surveillance by
biannual liver ultrasound with/without serum alpha-fetoprotein (AFP) for at-risk patients to achieve the goal of
detecting HCC at a curative stage. However, their accuracy remains relevantly low with sensitivity between 60-
70% at a specificity of 90%. As such, novel biomarkers for early detection of HCC are still desperately 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 and those cells within tumor microenvironment. Tumor-associated 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-associated EVs could mirror those of the parental tumor cells and those cells
within tumor microenvironment, exploiting the diagnostic potential of HCC-associated 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.
We propose an HCC EV Surface Protein Assay (SPA) to quantify subpopulations of HCC-associated EVs for
detecting early-stage HCC. The proposed HCC EV SPA couples two powerful technologies: Click Chemistry-
mediated EV Click Beads for isolating different subsets of HCC-associated EVs, and downstream 4-plex real-
time immuno-PCR for quantification of the isolated subsets of HCC EVs. One of the major challenges emerging
in the field of EV utilization for clinical use is the lack of robust and reproducible methods for the isolation of
subpopulations of tumor-associated EVs. Conventional methods for isolating EVs, such as ultracentrifugation,
filtration, and precipitation, are incapable of isolating subpopulation of tumor-associated EVs from total EVs. New
research efforts have been devoted to exploring immunoaffinity-based capture techniques for enriching tumor-
associated EVs from the plasma samples of patients with different solid tumors. However, there are challenges
identified for the single antibody-mediated tumor-derived EV enriching approaches, such as limited
sensitivity/specificity and a need for multiple capture antibodies to overcome the tumor heterogeneity. In order
to address these concerns, our research team developed HCC EV SPA, which combines a click chemistry-
mediated tumor-associated EV isolation, and downstream 4-plex real-time immuno-PCR. HCC EV SPA is
capable of highly sensitive and specific quantification of 32 subpopulations of HCC EVs in patients’ plasma
samples, based on the combined use of 8 different HCC-associated surface protein markers and four EV surface
markers. The long-term goal of this R01 proposal is to develop, refine, and validate the HCC EV SPA for
detecting early-stage HCC from at-risk liver cirrhotic patients by quantifying subpopulations of HCC EVs.
Terms: <ANX5><ANX5 Gene><ANXA5><ANXA5 gene><Address><Alcoholic Liver Diseases><Alpha-1-Fetoprotein><Alpha-Fetoglobulin><Anchorin CII><Anchorin CII Gene><Annexin A5 Gene><Annexin A5 Protein><Annexin V><Antibodies><Antigen Defined by Monoclonal Antibody AUAI><Assay><Bar Codes><Bio-Informatics><Bioassay><Bioinformatics><Biological Assay><Biology><Biometrics><Biometry><Biostatistics><Blood Plasma><Blood Serum><CBP-I><CD81><CD81 gene><Calphobindin I><Cancer Cause><Cancer Etiology><Case-Base Studies><Case-Comparison Studies><Case-Compeer Studies><Case-Referent Studies><Case-Referrent Studies><Case/Control Studies><Causality><Cell Body><Cells><Cessation of life><Chemistry><Chronic Hepatitis B><Circulation><Cirrhosis><Clinical><Clinical Practice Guideline><Couples><Coupling><DNA><Death><Deoxyribonucleic Acid><Development><Diagnosis><Diagnostic><Disease><Disorder><ENX2><ENX2 Gene><Early Diagnosis><Echography><Echotomography><Endonexin II><Endonexin II Gene><EpCAM><Epithelial Cellular Adhesion Molecule><Etiology><Exhibits><Fetuins><Filtration><Filtration Fractionation><GA733-2><GPC3><GPC3 gene><Gastrointestinal Tumor-Associated Antigen 2, 35-KD Glycoprotein><Glypican 3><Goals><Guidelines><HCC cell><HCC cell line><HCV infection><Hepatic Cirrhosis><Hepatic Disorder><Hepatitis B><Hepatitis C><Hepatitis C virus infection><Hepatitis, Viral, Non-A, Non-B, Parenterally-Transmitted><Hepatitus C><Hepatocarcinoma><Hepatocellular Carcinoma><Hepatocellular cancer><Hepatoma><Intratumoral heterogeneity><Joints><Lipocortin V Gene><Lipocortin-V><Liver><Liver Cells Carcinoma><Liver Cirrhosis><Liver diseases><Logistic Regressions><M4S1><MIC18><Mediating><Medical Ultrasound><Medical center><Membrane Component, Chromosome 4, Surface Marker 1><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Methods><Modality><Motivation><NAFLD><Oncology><Oncology Cancer><Operative Procedures><Operative Surgical Procedures><PAP-I><PP4 Gene><Pathology><Patients><Performance><Phosphatides><Phospholipids><Placental Anticoagulant Protein I><Placental Protein 4><Plasma><Plasma Serum><Precipitation><Primary Malignant Neoplasm of Liver><Primary carcinoma of the liver cells><Primary liver cancer><Prognosis><Protein Analysis><Quantitative Evaluations><Rat Homolog of OCI-5><Recommendation><Regression Analyses><Regression Analysis><Regression Diagnostics><Reproducibility><Research><Research Specimen><Reticuloendothelial System, Serum, Plasma><Risk><Sampling><Sensitivity and Specificity><Serum><Site><Solid Neoplasm><Solid Tumor><Specificity><Specimen><Statistical Regression><Surface><Surface Proteins><Surgical><Surgical Interventions><Surgical Procedure><TACSTD1><TACSTD1 gene><TAPA-1><TAPA1><TSPAN28><Techniques><Technology><Thromboplastin Inhibitor><Time><Training><Tumor Cell><Tumor-Associated Calcium Signal Transducer 1><Tumor-Derived><Ultracentrifugation><Ultrasonic Imaging><Ultrasonogram><Ultrasonography><Ultrasound Diagnosis><Ultrasound Medical Imaging><Ultrasound Test><VAC-Alpha><Validation><Vascular Anticoagulant-Alpha><Viral Hepatitis B><Viral hepatitis><alcohol induced hepatic injury><alcohol induced liver disorder><alcohol induced liver injury><alcohol related liver disease><alcohol-associated liver disease><alcohol-induced hepatic dysfunction><alcohol-induced liver disease><alcohol-induced liver dysfunction><alcohol-mediated liver dysfunction><alcohol-mediated liver injury><alcohol-related liver disease><alcoholic liver injury><annexin A5><assay development><barcode><cancer microenvironment><case-controlled studies><causation><cell type><chronic HBV infection><chronic hepatitis B virus infection><circulating biomarkers><circulating markers><cirrhotic><clinical practice and guidelines><cohort><computer based prediction><detection assay><developmental><diagnostic ability><diagnostic capability><diagnostic power><diagnostic ultrasound><diagnostic utility><diagnostic value><disease causation><early biomarkers><early detection><early detection biomarkers><early detection markers><ethanol induced hepatic injury><ethanol induced liver disorder><ethanol induced liver injury><ethanol liver disease><ethanol-induced hepatic dysfunction><ethanol-induced liver disease><ethanol-induced liver dysfunction><ethanol-mediated liver dysfunction><ethanol-mediated liver injury><extracellular vesicles><hep C><hepatic body system><hepatic disease><hepatic organ system><hepatitis non A non B><hepatitis virus infection><hepatocellular carcinoma cell line><hepatopathy><heterogeneity in tumors><in-vitro diagnostics><infection by hepatitis c virus><innovate><innovation><innovative><intra-tumoral heterogeneity><intratumor heterogeneity><lipid based nanoparticle><lipid nanoparticle><liver carcinoma><liver disorder><molecular pathology><neoplastic cell><new marker><non A, non B hepatitis><non-A, non-B hepatitis><non-alcohol fatty liver disease><non-alcoholic fatty liver disease><non-alcoholic liver disease><nonalcoholic fatty liver disease><novel biomarker><novel marker><particle><precipitations><predictive modeling><protein biomarkers><protein markers><serum hepatitis><sonogram><sonography><sound measurement><surgery><technology platform><technology system><tumor><tumor heterogeneity><tumor microenvironment><ultrasound><ultrasound imaging><ultrasound scanning><validations><α-Fetoproteins>