Fujiwara Naoto, Matsushita Yuki, Tempaku Mina, Tachi Yutaro, Kimura Genki, Izuoka Kiyora, Hayata Yuki, Kawamura Satoshi, Eguchi Akiko, Nakatsuka Takuma, Sato Masaya, Ono Atsushi, Murakami Eisuke, Tsuge Masataka, Oka Shiro, Hayashi Akinobu, Hirokawa Yoshifumi, Watanabe Masatoshi, Parikh Neehar D, Singal Amit G, Marrero Jorge A, Hoshida Yujin, Mizuno Shugo, Tateishi Ryosuke, Koike Kazuhiko, Fujishiro Mitsuhiro, Nakagawa Hayato
Department of Gastroenterology and Hepatology, Mie University, Mie, Japan.
Department of Gastroenterology, The University of Tokyo, Tokyo, Japan.
Hepatology. 2025 Apr 22. doi: 10.1097/HEP.0000000000001360.
Hepatic fiber morphology may significantly enhance our understanding of molecular alterations in metabolic dysfunction-associated steatotic liver disease (MASLD). We aimed to comprehensively characterize hepatic fiber morphological phenotypes in MASLD and their associated molecular alterations using multilayer omics analyses.
To quantify the morphological phenotypes of hepatic fibers, the artificial intelligence-based FibroNest algorithm (PharmaNest) was applied to 94 MASLD-affected liver biopsies, among which 12 (13%) had concurrent HCC. FibroNest identified 327 fiber phenotypes that were summarized into 8 major principal components, named FibroPC1-8. Next, molecular alterations captured by morphological fiber phenotypes were evaluated by comparison with genome-wide transcriptomics of paired liver samples. Pathway analyses revealed that FibroPCs more sensitively captured MASLD-related molecular alterations, such as upregulation of interleukin-6 and susceptibility to resmetirom, compared with the histological fibrosis stage. Among them, FibroPC4, which reflects reticular fibers, was associated with a gene signature predictive of incident HCC from MASLD. Furthermore, we used a spatial single-cell transcriptome, CosMx, to reveal the cell-cell interactions driving MASLD pathogenesis, as captured by FibroPC4. CosMx revealed that the FibroPC4-rich microenvironment contains HCC-promoting HSCs located adjacent to periportal endothelial cells. Neighboring cell analyses suggested that the HCC-promoting phenotype of HSCs was acquired by insulin growth factor-binding protein 7 secreted from senescent periportal endothelial cells. Consistently, in vitro experiments showed that insulin growth factor-binding protein 7 transformed HSCs into an HCC-promoting phenotype.
Hepatic morphological fiber phenotyping can reveal the disease progression and underlying mechanisms of MASLD.
肝纤维形态可能显著增进我们对代谢功能障碍相关脂肪性肝病（MASLD）分子改变的理解。我们旨在通过多层组学分析全面表征MASLD中肝纤维形态学表型及其相关分子改变。
为了量化肝纤维的形态学表型，基于人工智能的FibroNest算法（PharmaNest）应用于94例受MASLD影响的肝活检样本，其中12例（13%）同时患有肝癌。FibroNest识别出327种纤维表型，这些表型被归纳为8个主要主成分，命名为FibroPC1 - 8。接下来，通过与配对肝样本的全基因组转录组学进行比较，评估形态学纤维表型捕获的分子改变。通路分析显示，与组织学纤维化阶段相比，FibroPCs能更敏感地捕获与MASLD相关的分子改变，如白细胞介素 - 6的上调和对resmetirom的敏感性。其中，反映网状纤维的FibroPC4与预测MASLD患者发生肝癌的基因特征相关。此外，我们使用空间单细胞转录组CosMx来揭示由FibroPC4捕获的驱动MASLD发病机制的细胞间相互作用。CosMx显示，富含FibroPC4的微环境包含与门静脉周围内皮细胞相邻的促进肝癌的肝星状细胞（HSCs）。邻近细胞分析表明，HSCs促进肝癌的表型是由衰老的门静脉周围内皮细胞分泌的胰岛素生长因子结合蛋白7获得的。一致地，体外实验表明胰岛素生长因子结合蛋白7将HSCs转化为促进肝癌的表型。
肝形态学纤维表型分析可以揭示MASLD的疾病进展和潜在机制。
