Pharmacological Premature Termination Codon Readthrough of ABCB11 in Bile Salt Export Pump Deficiency: An In Vitro Study

• DOI: 10.1002/hep.31476
• Publication Date: 2020-07-23T20:31:45Z
• Authors: Rachida Amzal; Alice Thébaut; Martine Lapalus; Marion Almes; Brigitte Grosse; Elodie Mareux; Mauricette Collad...; Anne Davit‐Spraul; Laure Bidou; Olivier Namy; Emmanuel Jacquemin; Emmanuel Gonzales
• Keywords: HEK 293 cells; Ursodeoxycholic acid; Nonsense mutation

Progressive familial intrahepatic cholestasis type 2 (PFIC2) is a severe hepatocellular due to biallelic mutations in ABCB11 encoding the canalicular bile salt export pump (BSEP). Nonsense are responsible for most phenotypes. The aim was assess ability of drugs induce readthrough six nonsense (p.Y354X, p.R415X, p.R470X, p.R1057X, p.R1090X, and p.E1302X) identified patients with PFIC2.The G418, gentamicin, PTC124 studied using dual gene reporter system NIH3T3 cells. gentamicin lead expression full-length protein human embryonic kidney 293 (HEK293), HepG2, Can 10 cells immunodetection assays. function gentamicin-induced by measuring [3 H]-taurocholate transcellular transport stable Madin-Darby canine clones co-expressing Na+-taurocholate co-transporting polypeptide (Ntcp). Combinations chaperone (ursodeoxycholic acid, 4-phenylbutyrate [4-PB]) were investigated. In NIH3T3, aminoglycosides significantly increased level all studied, while only slightly p.E1302X. Gentamicin induced p.R1090X HEK293 resulting proteins localized within cytoplasm, except BsepR1090X , which also detected at plasma membrane HepG2 Additional treatment 4-PB ursodeoxycholic acid proportion clones, 40% increase transport, further additional treatment.This study constitutes proof concept therapy selected PFIC2 mutations.