McDaniel Kelly, Meng Fanyin, Wu Nan, Sato Keisaku, Venter Julie, Bernuzzi Francesca, Invernizzi Pietro, Zhou Tianhao, Kyritsi Konstantina, Wan Ying, Huang Qiaobing, Onori Paolo, Francis Heather, Gaudio Eugenio, Glaser Shannon, Alpini Gianfranco
Research, Central Texas Veterans Health Care System, Temple, TX.
Department of Medicine, Baylor Scott & White Health Digestive Disease Research Center, Texas A&M HSC and Baylor Scott & White Health, Temple, TX.
Hepatology. 2017 Feb;65(2):544-559. doi: 10.1002/hep.28831. Epub 2016 Nov 5.
Biliary-committed progenitor cells (small mouse cholangiocytes; SMCCs) from small bile ducts are more resistant to hepatobiliary injury than large mouse cholangiocytes (LGCCs) from large bile ducts. The definitive endoderm marker, forkhead box A2 (FoxA2), is the key transcriptional factor that regulates cell differentiation and tissue regeneration. Our aim was to characterize the translational role of FoxA2 during cholestatic liver injury. Messenger RNA expression in SMCCs and LGCCs was assessed by polymerase chain reaction (PCR) array analysis. Liver tissues and hepatic stellate cells (HSCs) from primary sclerosing cholangitis (PSC) and primary biliary cholangitis (PBC) patients were tested by real-time PCR for methylation, senescence, and fibrosis markers. Bile duct ligation (BDL) and multidrug resistance protein 2 (MDR2) knockout mice (MDR2 ) were used as animal models of cholestatic liver injury with or without healthy transplanted large or small cholangiocytes. We demonstrated that FoxA2 was notably enhanced in murine liver progenitor cells and SMCCs and was silenced in human PSC and PBC liver tissues relative to respective controls that are correlated with the epigenetic methylation enzymes, DNA methyltransferase (DNMT) 1 and DNMT3B. Serum alanine aminotransferase and aspartate aminotransferase levels in nonobese diabetic/severe combined immunodeficiency mice engrafted with SMCCs post-BDL showed significant changes compared to vehicle-treated mice, along with improved liver fibrosis. Enhanced expression of FoxA2 was observed in BDL mouse liver after SMCC cell therapy. Furthermore, activation of fibrosis signaling pathways were observed in BDL/MDR2 mouse liver as well as in isolated HSCs by laser capture microdissection, and these signals were recovered along with reduced hepatic senescence and enhanced hepatic stellate cellular senescence after SMCC engraft.
The definitive endoderm marker and the positive regulator of biliary development, FoxA2, mediates the therapeutic effect of biliary-committed progenitor cells during cholestatic liver injury. (Hepatology 2017;65:544-559).
来自小胆管的胆管定向祖细胞（小鼠小胆管细胞；SMCCs）比来自大胆管的大鼠胆管细胞（LGCCs）对肝胆损伤更具抗性。确定的内胚层标志物叉头框A2（FoxA2）是调节细胞分化和组织再生的关键转录因子。我们的目的是表征FoxA2在胆汁淤积性肝损伤中的翻译作用。通过聚合酶链反应（PCR）阵列分析评估SMCCs和LGCCs中的信使核糖核酸表达。通过实时PCR检测原发性硬化性胆管炎（PSC）和原发性胆汁性胆管炎（PBC）患者的肝组织和肝星状细胞（HSCs）中的甲基化、衰老和纤维化标志物。胆管结扎（BDL）和多药耐药蛋白2（MDR2）敲除小鼠（MDR2-/-）被用作胆汁淤积性肝损伤的动物模型，有无健康移植的大小胆管细胞。我们证明，相对于与表观遗传甲基化酶DNA甲基转移酶（DNMT）1和DNMT3B相关的各自对照，FoxA2在小鼠肝祖细胞和SMCCs中显著增强，而在人PSC和PBC肝组织中沉默。与载体处理的小鼠相比，BDL后植入SMCCs的非肥胖糖尿病/严重联合免疫缺陷小鼠的血清丙氨酸转氨酶和天冬氨酸转氨酶水平有显著变化，同时肝纤维化得到改善。SMCC细胞治疗后，BDL小鼠肝脏中观察到FoxA2表达增强。此外，通过激光捕获显微切割在BDL/MDR2小鼠肝脏以及分离的HSCs中观察到纤维化信号通路的激活，并且在植入SMCCs后，这些信号随着肝衰老的减少和肝星状细胞衰老的增强而恢复。
确定的内胚层标志物和胆管发育的正调节因子FoxA2在胆汁淤积性肝损伤期间介导胆管定向祖细胞的治疗作用。（《肝脏病学》2017年；65:544 - 559）
