In vitro recovery of FIX clotting activity as a marker of highly functional hepatocytes in a hemophilia B iPSC model

• DOI: 10.1002/hep.32211
• Publication Date: 2021-10-23T07:25:09Z
• Authors: Eléanor Luce; Clara Steichen; Mickaël Allouche; Antonietta Messina; Jean‐Marie Heslan; Thierry Lambert; Anne Weber; Tuan Huy Nguyen; Olivier Christophe; Anne Dubart‐Kuppe...
• Keywords: Clotting factor

Abstract Background and Aims Pluripotent stem cell–derived hepatocytes differentiated in monolayer culture are known to have more fetal than adult hepatocyte characteristics. If numerous studies tend show that this immature phenotype might not necessarily be an obstacle their use transplantation, other applications such as drug screening, toxicological studies, or bioartificial livers reliant on functionality require full differentiation of hepatocytes. New technologies been used improve the process recent years, usually evaluated by measuring albumin production CYP450 activity. Here we complex most importantly activity coagulation factor IX (FIX) produced mature assess hemophilia B (HB) patient’s induced pluripotent cells (iPSCs) both organoids. Approach Results Indeed, HB is X‐linked monogenic disease due impaired FIX synthesized liver. We developed vitro model using iPSCs generated from fibroblasts a severe patient. CRISPR/Cas9 technology target genomic insertion 9 minigene bearing Padua mutation enhance Noncorrected corrected were into under two‐dimensional three‐dimensional protocols deciphered active vitro. Finally, assessed therapeutic efficacy approach vivo mouse HB. Conclusions Functional FIX, whose post‐translational modifications only occur fully hepatocytes, was Immunohistochemistry analyses indicated good cell engraftment, detected plasma transplanted animals confirmed rescue bleeding phenotype.