Wu Xiaoshan, Jiang Dacheng, Wang Yuchen, Li Xin, Liu Chenyu, Chen Yanhao, Sun Wei, He Ruikun, Yang Yi, Gu Xiaosong, Jiang Chunping, Ding Qiurong
CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, P.R. China.
Optogenetics and Synthetic Biology Interdisciplinary Research Center, School of Biotechnology, East China University of Science and Technology, Shanghai, P.R. China.
Hepatol Commun. 2024 Nov 29;8(12). doi: 10.1097/HC9.0000000000000585. eCollection 2024 Dec 1.
Metabolic-associated steatohepatitis (MASH) is one of the most prevalent liver diseases worldwide, with a global prevalence estimated between 3% and 5%, posing a significant health burden. Human liver organoids (HLOs) have previously been generated to model steatohepatitis, offering a potential cellular disease model for studying MASH. However, the current HLO model lacks detailed molecular characterizations and requires further improvement.
HLOs derived from human pluripotent stem cells were treated with oleic acid and TGFβ to mimic the MASH progression. Treated HLOs were then analyzed using both bulk and single-cell RNA sequencing. Functional characterization was performed through staining with BODIPY, TMRM, CellROX, and Collagen I, as well as terminal deoxynucleotidyl transferase dUTP nick end labeling and ELISA assays. In addition, a test using the MASH HLO model to validate the hepatoprotective effects of several herb extracts was also conducted.
Both RNA-seq and single-cell RNA sequencing demonstrated a close resemblance of multiple molecular signatures and key intercellular communications in and between hepatocyte-like cells and stellate-like cells in the MASH HLO model, compared to human MASH. Furthermore, functional characterizations revealed progressive features of human MASH in the MASH HLO model, including severe steatosis, oxidative stress, mitochondrial dysfunction, inflammation, and fibrosis. In addition, the Schisandra extracts have been demonstrated to have significant antioxidative, anti-inflammatory, and antifibrotic properties in the context of MASH.
This study offers an improved HLO disease model of human MASH, which can be potentially applied to facilitate the understanding of the MASH pathogenesis and the discovery of effective treatments.
代谢相关脂肪性肝炎（MASH）是全球最常见的肝脏疾病之一，全球患病率估计在3%至5%之间，造成了重大的健康负担。此前已生成人类肝脏类器官（HLOs）来模拟脂肪性肝炎，为研究MASH提供了一种潜在的细胞疾病模型。然而，当前的HLO模型缺乏详细的分子特征，需要进一步改进。
用油酸和转化生长因子β处理源自人类多能干细胞的HLOs，以模拟MASH的进展。然后使用批量和单细胞RNA测序对处理后的HLOs进行分析。通过用BODIPY、TMRM、CellROX和I型胶原蛋白染色，以及末端脱氧核苷酸转移酶dUTP缺口末端标记和ELISA测定进行功能表征。此外，还进行了一项使用MASH HLO模型验证几种草药提取物肝脏保护作用的试验。
与人类MASH相比，RNA测序和单细胞RNA测序均表明，MASH HLO模型中肝细胞样细胞和星状细胞样细胞内以及它们之间的多种分子特征和关键细胞间通讯非常相似。此外，功能表征揭示了MASH HLO模型中人类MASH的进展特征，包括严重脂肪变性、氧化应激、线粒体功能障碍、炎症和纤维化。此外，已证明五味子提取物在MASH背景下具有显著的抗氧化、抗炎和抗纤维化特性。
本研究提供了一种改进的人类MASH的HLO疾病模型，可潜在地用于促进对MASH发病机制的理解和有效治疗方法的发现。
