title: Development of an in vitro model to test antifibrotic drugs on primary human liver myofibroblasts.
abstract: We have developed a culture model to assess antifibrotic drugs using normal human liver myofibroblasts (HLMFs) obtained from 31 subjects. Activation was evaluated in terms of α-smooth muscle actin (α-SMA) and collagen 1 (Coll1) expression using RT-PCR, and proliferation as the uptake of 5-ethynil-2'-deoxyuridine. Under analysis of variance, between-subject differences accounted for 70% of all variability and inter-experiment differences for 30%. The sensitivity of the model was determined by quantifying the effects in terms of relative expression, which were 0.74±0.03 for cyclosporine A (CsA) and 2.4±0.10 for transforming growth factor-beta (TGF-β) (P<0.0001 vs no treatment) for α-SMA expression. Inter-subject variations in α-SMA and Coll1 expression enabled the classification of subjects as potentially low or high fibrosers. Finally, we observed that pirfenidone (which has beneficial effects in vivo) significantly reduced the expressions of α-SMA and Coll1, whereas the angiotensin-converting enzyme inhibitor losartan (which has no effect in vivo) had no significant effect. Our model may thus detect the antifibrotic properties of drugs. Antifibrotic drugs with promising clinical relevance could possibly be selected using a bank of HLMFs from high fibrosers.
authors.affiliation: Human HepCell, Faculté de Médecine Pierre et Marie Curie, Site Saint-Antoine, Paris, France.|Sorbonne Universités, UPMC University Paris 06, Paris, France.|INSERM, UMR_S 938, CDR Saint-Antoine, Paris, France.
authors.forename: Lynda
authors.initials: L
authors.lastname: Aoudjehane
authors.affiliation: AP-HP, Hôpital Saint Antoine, Unité de Biostatistique, Paris, France.
authors.forename: Pierre-Yves
authors.initials: PY
authors.lastname: Boelle
authors.affiliation: Human HepCell, Faculté de Médecine Pierre et Marie Curie, Site Saint-Antoine, Paris, France.
authors.forename: Grégoire
authors.initials: G
authors.lastname: Bisch
authors.affiliation: Sorbonne Universités, UPMC University Paris 06, Paris, France.|INSERM, UMR_S 938, CDR Saint-Antoine, Paris, France.
authors.forename: Rolland
authors.initials: R
authors.lastname: Delelo
authors.affiliation: Sorbonne Universités, UPMC University Paris 06, Paris, France.|AP-HP, Hôpital Saint Antoine, Service de Chirurgie Digestive, Paris, France.
authors.forename: François
authors.initials: F
authors.lastname: Paye
authors.affiliation: Sorbonne Universités, UPMC University Paris 06, Paris, France.|INSERM, UMR_S 938, CDR Saint-Antoine, Paris, France.|AP-HP, Hôpital Pitié-Salpêtrière, Servie de Chirurgie Digestive et Transplantation Hépatique, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Scatton
authors.affiliation: Sorbonne Universités, UPMC University Paris 06, Paris, France.|INSERM, UMR_S 938, CDR Saint-Antoine, Paris, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Housset
authors.affiliation: Human HepCell, Faculté de Médecine Pierre et Marie Curie, Site Saint-Antoine, Paris, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Becquart
authors.affiliation: Sorbonne Universités, UPMC University Paris 06, Paris, France.|INSERM, UMR_S 938, CDR Saint-Antoine, Paris, France.|AP-HP, Hôpital Pitié-Salpêtrière, Unité de Transplantation Hépatique, Paris, France.
authors.forename: Yvon
authors.initials: Y
authors.lastname: Calmus
authors.affiliation: Sorbonne Universités, UPMC University Paris 06, Paris, France.|INSERM, UMR_S 938, CDR Saint-Antoine, Paris, France.|AP-HP, Hôpital Pitié-Salpêtrière, Unité de Transplantation Hépatique, Paris, France.
authors.forename: Filomena
authors.initials: F
authors.lastname: Conti
mesh_terms: D000199:Actins / Q000235:genetics; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D008103:Liver Cirrhosis / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D019808:Losartan / Q000494:pharmacology; D008954:Models, Biological; D058628:Myofibroblasts / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D011728:Pyridones / Q000494:pharmacology
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D000894:Anti-Inflammatory Agents, Non-Steroidal; D024042:Collagen Type I; D011728:Pyridones; C093844:pirfenidone; D019808:Losartan