Clinical Validation of Peripheral Blood Mononuclear Cell DNA Methylation Markers for accurate Early Detection of Hepatocellular Carcinoma

Early detection of Hepatocellular carcinoma (HCC) has a dramatic effect on the outcome; there is therefore a dire need for non-invasive blood-based markers for early detection of HCC. The immune system is impacted by HCC and therefore peripheral blood cells (PBMC) could potentially provide a non-invasive sample for detection of cancer. DNA methylation is an epigenetic mark on DNA that is dramatically altered in cancers, but it is still not well established whether changes in DNA methylation in white blood cells could be used as biomarkers for early detection of cancer. Our prior work examining a cohort of healthy, chronic hepatitis and HCC from China unveiled DNA methylation profiles in blood PBMC that differentiated even early stages of HCC from chronic hepatitis B and C and healthy controls. The clinical study reported here validated these findings using a different method of evaluating DNA methylation in a larger and independent cohort from a different ethnicity from Bangladesh, focusing on the specificity and sensitivity of early-stage HCC detection. We created integrative scores of shortlisted regions (M8 and M4), that enhanced the detection efficacy for HCC, especially in early stages. When these scores were amalgamated with the previously developed cell free DNA epiLiver marker and alpha fetal protein (AFP) antigen, there was a notable improvement in sensitivity across all HCC stages, particularly in the early-stage detection. Our findings highlight the significant advancement in early-stage HCC detection through the combination of gene methylation profiles with existing markers, leading to the development of a novel diagnostic tool. This approach not only paves the way for timely clinical interventions with its non-invasive nature but also sets a new standard in sensitivity and specificity, as evidenced by our comprehensive comparative analysis, marking a pivotal breakthrough in non-invasive HCC diagnostics.