Lapalus Martine, Mareux Elodie, Amzal Rachida, Drège Emmanuelle, Riahi Yosra, Petit Sylvain, Banet Manon, Falguières Thomas, Callebaut Isabelle, Figadère Bruno, Joseph Delphine, Gonzales Emmanuel, Jacquemin Emmanuel
Université Paris-Saclay, Inserm, Physiopathogénèse et Traitement des Maladies du Foie, FHU Hepatinov, 91400 Orsay, France.
Université Paris-Saclay, CNRS, BioCIS, 91400 Orsay, France.
Int J Mol Sci. 2025 May 29;26(11):5232. doi: 10.3390/ijms26115232.
Progressive familial intrahepatic cholestasis type 2 (PFIC2) is a severe hepatocellular cholestasis due to biallelic variations in the (ATP-binding cassette B11) gene encoding the canalicular bile salt export pump (BSEP). Some missense variants identified in patients with PFIC2 do not traffic properly to the canalicular membrane. However, 4-phenybutyrate (4-PB) has been shown in vitro to partially correct the mis-trafficking of selected variants, resulting in an improvement of the medical conditions of corresponding PFIC2 patients. Herein, we report the ability of 4-PB analogous or homologous drugs and of non-4-PB related chemical correctors to rescue the canalicular expression and the activity of the folding-defective Abcb11 variant. New compounds, either identified by screening a chemical library or designed by structural homology with 4-PB (or its metabolites) and synthesized, were evaluated in vitro for their ability to (i) correct the canalicular localization of Abcb11 after transfection in hepatocellular polarized cell lines; (ii) restore the H-taurocholate transport of the Abcb11 protein in Madin-Darby canine kidney (MDCK) cells stably co-expressing Abcb11 and the sodium taurocholate co-transporting polypeptide (Ntcp/). Glycerol phenylbutyrate (GPB), phenylacetate (PA, the active metabolite of 4-PB), 3-hydroxy-2-methyl-4-phenylbutyrate (HMPB, a 4-PB metabolite analog chemically synthesized in our laboratory) and 4-oxo-1,2,3,4-tetrahydro-naphthalene-carboxylate (OTNC, from the chemical library screening) significantly increased the proportion of canalicular Abcb11 protein. GPB, PA, ursodeoxycholic acid (UDCA), alone or in combination with 4-PB, suberoylanilide hydroxamic acid (SAHA), C18, VX-445, and/or VX-661, significantly corrected both the traffic and the activity of Abcb11. Such correctors could represent new pharmacological insights for improving the condition of patients with ABCB11 deficiency due to missense variations affecting the transporter's traffic.
2型进行性家族性肝内胆汁淤积症（PFIC2）是一种严重的肝细胞性胆汁淤积症，由编码胆小管胆汁盐输出泵（BSEP）的ABCB11基因双等位基因变异引起。在PFIC2患者中鉴定出的一些错义变体不能正常转运至胆小管膜。然而，4-苯丁酸（4-PB）在体外已显示可部分纠正选定变体的错误转运，从而改善相应PFIC2患者的病情。在此，我们报告了4-PB类似物或同源药物以及非4-PB相关化学校正剂挽救折叠缺陷型Abcb11变体的胆小管表达和活性的能力。通过筛选化学文库鉴定或通过与4-PB（或其代谢物）进行结构同源性设计并合成的新化合物，在体外评估其（i）在肝细胞极化细胞系中转染后纠正Abcb11胆小管定位的能力；（ii）在稳定共表达Abcb11和牛磺胆酸钠共转运多肽（Ntcp）的Madin-Darby犬肾（MDCK）细胞中恢复Abcb11蛋白的牛磺胆酸转运能力。甘油苯丁酸酯（GPB）、苯乙酸（PA，4-PB的活性代谢物）、3-羟基-2-甲基-4-苯丁酸（HMPB，我们实验室化学合成的4-PB代谢物类似物）和4-氧代-1,2,3,4-四氢萘甲酸酯（OTNC，来自化学文库筛选）显著增加了胆小管Abcb11蛋白的比例。GPB、PA、熊去氧胆酸（UDCA）单独或与4-PB、辛二酰苯胺异羟肟酸（SAHA）、C18、VX-445和/或VX-661联合使用，均显著纠正了Abcb11的转运和活性。此类校正剂可能为改善因影响转运蛋白转运的错义变异导致ABCB11缺乏患者的病情提供新的药理学见解。
