Department of Pharmaceutical Sciences, College of Pharmacy and Pharmaceutical Sciences, Washington State University Health Sciences, Spokane, WA 99202, USA.
Genes (Basel). 2024 Mar 13;15(3):357. doi: 10.3390/genes15030357.
Non-alcoholic steatohepatitis (NASH, also known as MASH) is a severe form of non-alcoholic fatty liver disease (NAFLD, also known as MASLD). Emerging data indicate that the progression of the disease to MASH is higher in postmenopausal women and that genetic susceptibility increases the risk of MASH-related cirrhosis. This study aimed to investigate the association between genetic polymorphisms in MASH and sexual dimorphism. We applied whole-exome sequencing (WES) to identify gene variants in 8 age-adjusted matched pairs of livers from both male and female patients. Sequencing alignment, variant calling, and annotation were performed using standard methods. Polymerase chain reaction (PCR) coupled with Sanger sequencing and immunoblot analysis were used to validate specific gene variants. cBioPortal and Gene Set Enrichment Analysis (GSEA) were used for actionable target analysis. We identified 148,881 gene variants, representing 57,121 and 50,150 variants in the female and male cohorts, respectively, of which 251 were highly significant and MASH sex-specific ( < 0.0286). Polymorphisms in , , , , , , and were highly expressed in male liver samples. In female samples, Polymorphisms in , , , , , , and were identified. A heterozygous variant 1151G>T located on 18q21.32 for (rs3809983) was validated by Sanger sequencing and expressed only in female samples. Immunoblot analysis confirmed that the protein level of β-catenin in female samples was 2-fold higher than normal, whereas ALPK2 expression was 0.5-fold lower than normal. No changes in the protein levels of either ALPK2 or β-catenin were observed in male samples. Our study suggests that the perturbation of canonical Wnt/β-catenin signaling observed in postmenopausal women with MASH could be the result of polymorphisms in .
非酒精性脂肪性肝炎（NASH，也称为 MASH）是一种严重的非酒精性脂肪性肝病（NAFLD，也称为 MASLD）。新出现的数据表明，绝经后妇女患 MASH 的疾病进展更高，遗传易感性增加了与 MASH 相关的肝硬化的风险。本研究旨在探讨 MASH 基因多态性与性别二态性之间的关联。我们应用全外显子组测序（WES）在 8 对年龄匹配的男性和女性患者的肝脏中鉴定基因变异。采用标准方法进行测序比对、变异调用和注释。聚合酶链反应（PCR）与 Sanger 测序和免疫印迹分析用于验证特定基因变异。cBioPortal 和基因集富集分析（GSEA）用于可操作的目标分析。我们鉴定了 148881 个基因变异，分别代表女性和男性队列中的 57121 和 50150 个变异，其中 251 个变异高度显著且为 MASH 性别特异性（<0.0286）。在男性肝组织中，、、、、、、和 中的多态性高度表达。在女性样本中，、、、、、、和 中的多态性被鉴定出来。在 18q21.32 上位于 的杂合子变异 1151G>T （rs3809983） 对于 （rs3809983），通过 Sanger 测序进行了验证，并且仅在女性样本中表达。免疫印迹分析证实，女性样本中 β-catenin 的蛋白水平是正常的 2 倍，而 ALPK2 的表达水平是正常的 0.5 倍。在男性样本中，没有观察到 ALPK2 或 β-catenin 的蛋白水平发生变化。我们的研究表明，绝经后妇女 MASH 中观察到的经典 Wnt/β-catenin 信号通路的扰动可能是由于 中的多态性所致。
