Transcriptomic signature of treatment with recombinant laminin 511-E8 and acetazolamide in human H69 cholangiocytes

IgG4-related cholangitis (IRC) is the hepatobiliary manifestation of IgG4-related disease (IgG4-RD), a systemic, B-cell driven, fibroinflammatory disorder. To date, numerous autoantigens have been described including laminin 511-E8 and carbonic anhydrase (CA) enzymes (PMID 30089633, 15647194). Laminin 511-E8 is an extracellular matrix protein that has been shown to upregulate secretory components and aid in the cholangiocyte differentiation of induced pluripotent stem cells (PMID 27103433). Carbonic anhydrase enzymes catalyse the reversible hydration of carbon dioxide, thereby producing bicarbonate and a single proton. In humans there are 14 different CA isoforms with a confined subcellular distribution. All isoforms can be targeted by the pan-CA inhibitor acetazolamide, which is a registered drug for multiple indications. We aimed to investigate the role of IgG4-RD autoantigens laminin 511-E8 and CA enzymes in human cholangiocytes. Transcriptional profiling may provide insights into the underlying molecular mechanisms of these proteins on the cholangiocellular level. Thus, by RNA sequencing the transcriptomic signature of recombinant laminin 511-E8 and acetazolamide treatment was assessed in human H69 cholangiocytes. RNA quality was assessed for four biological replicates of each group by RIN scores. For each group the three samples with the highest RIN scores were selected for library preparation and mRNA sequencing. Group names are Ctrl H69, Laminin 511-E8 0.25ug/cm2 H69 and Acetazolamide 10uM H69. Dose titration experiments and available literature were taken into account to arrive at the chosen experimental concentrations.