The goal of the proposed work is development of sensitive diagnostic serological assay panels for detection of hepatocellular carcinoma (HCC). Currently, the only biomarker used in HCC diagnosis is the measurement of alpha feto protein (AFP) in patient serum, which exhibits poor sensitivity and specificity. There is a need for alternative biomarkers to more efficiently diagnose HCC, particularly for early detection, when aggressive treatments would be most effective. Approaches that detect autoimmune responses to cancer-related antigens are being developed as an effective means of early cancer detection, potentially detectable before measurable levels of the cancer antigens themselves are present in serum. Our long term goal is to develop commercial clinical diagnostic multiplex assays for early detection and risk stratification of hepatocellular carcinoma. In this pilot study, we will optimize the use of Meso Scale Diagnostics (MSD) sensitive Multi-Array technology for an efficient and sensitive detection of autoantibodies identified previously in serum of HCC patients using protein arrays by Dr. Goldman's group at Georgetown University. The optimized assays will be tested in a pilot study of 100 samples of HCC patient and controls with chronic liver disease (CLD) from Egypt and 50 patients and controls with CLD from the US. Validation of the autoantibodies associated with progression of liver disease to HCC is expected to generate new hypotheses for targeted disease management. Multiple distinct antigens of interest will be immobilized in MSD Multi- Array panels, which can accommodate 1-25 assays/well of a 96-well plate. The assay panel will be optimized for ability to detect specific humoral responses in HCC patient samples, allowing multiple simultaneous determinations per well. MSD multiplex assay panels have shown sensitivities as low as 0.1 pg/mL for serum proteins, 3-5 orders of magnitude dynamic range, very high throughput, and minimal sample usage (25 microliters/well of diluted sample), factors which are critical in developing successful serological screening panels for the proposed study. We will further validate our findings on a larger population in phase II of the project. The phase II study will also expand the detection to autoantibodies associated with pre-malignant stages of liver disease progression. Defining clinically applicable tests for the detection of early stage cancer has potentially far reaching consequences for disease management and patient health. The proposed academic/industry collaboration addresses NCI goals of translational medicine, by advancing potential biomarkers from discovery towards clinical applications using a versatile and robust assay platform.
The proposed studies aim to develop screening panels for sensitive and specific detection of hepatocellular carcinoma by measuring the immune response of patients to proteins derived from the developing cancer, a more sensitive means of early cancer detection than directly measuring levels of cancer-related proteins themselves. Efficient early detection would lead to increased survival rates for HCC, particularly in high risk populations (patients with chronic liver diseases). The sensitive MSD technology that will be used allows the detection of several immune responses simultaneously, providing more accurate disease detection capability and will eventually help to translate discoveries from research laboratories into medical practice. ? ? ?
