Division of Gastroenterology, Hepatology and Nutrition of Cincinnati Children's Hospital Medical Center and the Department of Pediatrics of the University of Cincinnati College of Medicine, Cincinnati, Ohio.
Division of Gastroenterology, Hepatology and Nutrition of Cincinnati Children's Hospital Medical Center and the Department of Pediatrics of the University of Cincinnati College of Medicine, Cincinnati, Ohio.
Gastroenterology. 2019 Oct;157(4):1138-1152.e14. doi: 10.1053/j.gastro.2019.06.017. Epub 2019 Jun 19.
BACKGROUND & AIMS: Little is known about the factors that affect outcomes of patients with biliary atresia and there are no medical therapies that increase biliary drainage.
Liver biopsies and clinical data were obtained from infants with cholestasis and from children without liver disease (controls); messenger RNA (mRNA) was isolated, randomly assigned to discovery (n = 121) and validation sets (n = 50), and analyzed by RNA sequencing. Using the Superpc R package followed by Cox regression analysis, we sought to identify gene expression profiles that correlated with survival without liver transplantation at 24 months of age. We also searched for combinations of gene expression patterns, clinical factors, and laboratory results obtained at diagnosis and at 1 and 3 months after surgery that associated with transplant-free survival for 24 months of age. We induced biliary atresia in BALB/c mice by intraperitoneal administration of Rhesus rotavirus type A. Mice were given injections of the antioxidants N-acetyl-cysteine (NAC) or manganese (III) tetrakis-(4-benzoic acid)porphyrin. Blood and liver tissues were collected and analyzed by histology and immunohistochemistry.
We identified a gene expression pattern of 14 mRNAs associated with shorter vs longer survival times in the discovery and validation sets (P < .001). This gene expression signature, combined with level of bilirubin 3 months after hepatoportoenterostomy, identified children who survived for 24 months with an area under the curve value of 0.948 in the discovery set and 0.813 in the validation set (P < .001). Computer models correlated a cirrhosis-associated transcriptome with decreased times of transplant-free survival; this transcriptome included activation of genes that regulate the extracellular matrix and numbers of activated stellate cells and portal fibroblasts. Many mRNAs expressed at high levels in liver tissues from patients with 2-year transplant-free survival had enriched scores for glutathione metabolism. Among mice with biliary atresia given injections of antioxidants, only NAC reduced histologic features of liver damage and serum levels of aminotransferase, gamma-glutamyl transferase, and bilirubin. NAC also reduced bile duct obstruction and liver fibrosis and increased survival times.
In studies of liver tissues from infants with cholestasis, we identified a 14-gene expression pattern that associated with transplant-free survival for 2 years. mRNAs encoding proteins that regulate fibrosis genes were increased in liver tissues from infants who did not survive for 2 years, whereas mRNAs that encoded proteins that regulate glutathione metabolism were increased in infants who survived for 2 years. NAC reduced liver injury and fibrosis in mice with biliary atresia, and increased survival times. Agents such as NAC that promote glutathione metabolism might be developed for treatment of biliary atresia.
目前对于影响胆道闭锁患儿结局的因素知之甚少，也没有能够增加胆汁引流的医学治疗方法。
收集胆汁淤积患儿和无肝脏疾病儿童（对照组）的肝活检和临床资料；分离信使 RNA（mRNA），随机分配到发现（n=121）和验证集（n=50），并通过 RNA 测序进行分析。使用 Superpc R 软件包和 Cox 回归分析，我们试图确定与 24 月龄时无肝移植生存相关的基因表达谱。我们还寻找了在诊断时、手术后 1 个月和 3 个月获得的基因表达模式、临床因素和实验室结果的组合，这些组合与 24 月龄时的无移植生存相关。我们通过腹腔内给予恒河猴轮状病毒 A 型诱导 BALB/c 小鼠发生胆道闭锁。给小鼠注射抗氧化剂 N-乙酰半胱氨酸（NAC）或锰（III）四（4-苯甲酸）卟啉。收集血液和肝脏组织，通过组织学和免疫组织化学进行分析。
我们在发现和验证集中确定了与较短和较长生存时间相关的 14 个 mRNA 的基因表达模式（P<0.001）。该基因表达特征与肝门空肠吻合术后 3 个月的胆红素水平相结合，在发现集中的曲线下面积值为 0.948，在验证集中为 0.813，可识别出 24 月龄时存活的儿童（P<0.001）。计算机模型将与肝硬化相关的转录组与无移植生存时间缩短相关联；该转录组包括调节细胞外基质和激活的星状细胞和门脉成纤维细胞数量的基因的激活。在 2 年无移植生存的患者肝脏组织中表达水平较高的许多 mRNAs 具有丰富的谷胱甘肽代谢评分。在接受抗氧化剂注射的胆道闭锁小鼠中，只有 NAC 降低了肝损伤和血清转氨酶、γ-谷氨酰转移酶和胆红素的组织学特征。NAC 还减少了胆管阻塞和肝纤维化，并延长了生存时间。
在对胆汁淤积婴儿的肝脏组织进行的研究中，我们确定了与 2 年无移植生存相关的 14 个基因表达模式。在未存活 2 年的婴儿的肝脏组织中，编码调节纤维化基因的蛋白质的 mRNAs 增加，而编码调节谷胱甘肽代谢的蛋白质的 mRNAs 在存活 2 年的婴儿中增加。NAC 降低了胆道闭锁小鼠的肝损伤和纤维化，并延长了生存时间。可以开发促进谷胱甘肽代谢的 NAC 等药物来治疗胆道闭锁。
