VEGFA mRNA-LNP promote biliary epithelial cell-to-hepatocyte conversion in acute and chronic liver diseases and reverses steatosis and fibrosis

The goal of this experiment was to analyse transcriptomic changes between Biliary epithelial cells or BECs from the liver of mice treated with Acetaminophen (a drug that causes acute liver injury) in combination with AAV8-Tbg-p21 viral vector to induce hepatocyte senecence. The aim was to understand the impact of injury on transcriptional status of biliary epithelial cells. Briefly, we analyzed mouse liver cells (hepatocytes and non-parenchymal cells fractions) obtained by in-situ liver perfusion 48 h after injury. We further extracted cells corresponding to the hepatocyte and biliary epithelial cell gene signature for detailed analysis. Two separate clusters of BECs (BEC1 and BEC2) were further identified and analyzed to study their transcriptional changes and corresponding pathways. Two male mice are treated with AAV8-Tbg-p21 viral vectors one week prior to injection with acute liver injury causing drug, acetaminophen. 48 h after injury, liver is perfused in-situ and digested to obtain liver cells as two fractions. In the first step, hepatocyte fraction is recovered which is composed of predominantly hepatocytes and also some non-parenchymal cells. This was followed by subsequent liver digestion to procure non-parenchymal cells (NPCs) fraction consisting of blood cells, immune cells, cholangiocytes, stellate cells as well as smaller hepatocytes. Both hepatocytes and NPCs fractions from each mouse were harvested and submitted separately for library preparation.