A novel and inexpensive multiplex miRNA platform for liver diseases screening

Abstract Alcohol-associated liver disease (AALD) is a major driver of global liver-related morbidity and mortality. Alcohol alone accounts for 30% to 50% of cirrhosis‐related deaths globally. At least half of people living with type 2 diabetes have nonalcoholic fatty liver disease and are at higher risk to develop nonalcoholic or alcohol associated steatohepatitis and cirrhosis. miRNA biomarker characterization is an emerging field for early detection, monitoring and the evaluation of treatment response in disease conditions and clinical applications of miRNA-based biomarkers is rapidly growing. Recently, miRNA-based diagnostics assays are being offered in clinical oncology that use widely available RT-qPCR methodology. However, these assays are low throughput, and high turnaround time, due to ability to perform analysis of only one miRNA/reaction. To date, there is no qPCR-based method available that can detect multiple miRNAs in a single qPCR reaction. KASA BIO is developing Zip-Melt Multiplex miRNA Solution that enables reverse transcription and detection of multiple miRNAs of interest. This novel solution overcomes the limitations of current technologies for miRNA detection using RT-qPCR methodology. Our proposed solution can detect multiple miRNAs from various sample matrices (tissue or body fluids) and will provide a cost-effective, eco-friendly technology to meet the clinical requirement of multianalyte detection in a single workflow. Compared to the existing kits, the advantage of our solution is that it is easy to use at various research settings including resource limited setting such as point of care. This project proposes to develop Zip-MeltTM HepmiR Assay to determine analytical performance and equivalence of Zip-Melt TM Multiplex miRNA Solution in comparison with commercially available RT-qPCR Array and to use the Assay in clinical research to determine miRNA signatures important for liver diseases (screening or diagnosis or monitoring). The proposed technology will have a significant impact on translational research by providing an affordable multiplex miRNA qPCR solution for life sciences research and will address the molecular diagnostic need.