Department of Chemistry and Molecular Biology, Sahlgrenska University Hospital, University of Gothenburg, Gothenburg, Sweden.
Department of Biochemistry, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
Hepatol Commun. 2023 Mar 17;7(4). doi: 10.1097/HC9.0000000000000037. eCollection 2023 Apr 1.
NAFLD has become the leading cause of chronic liver disease worldwide afflicting about one quarter of the adult population. NASH is a severe subtype of NAFLD, which in addition to hepatic steatosis connotes liver inflammation and hepatocyte ballooning. In light of the exponentially increasing prevalence of NAFLD, it is imperative to gain a better understanding of its molecular pathogenesis. The aim of this study was to examine the potential role of STE20-type kinase TAOK1 -a hepatocellular lipid droplet-associated protein-in the regulation of liver lipotoxicity and NAFLD etiology.
The correlation between TAOK1 mRNA expression in liver biopsies and the severity of NAFLD was evaluated in a cohort of 62 participants. Immunofluorescence microscopy was applied to describe the subcellular localization of TAOK1 in human and mouse hepatocytes. Metabolic reprogramming and oxidative/endoplasmic reticulum stress were investigated in immortalized human hepatocytes, where TAOK1 was overexpressed or silenced by small interfering RNA, using functional assays, immunofluorescence microscopy, and colorimetric analysis. Migration, invasion, and epithelial-mesenchymal transition were examined in TAOK1-deficient human hepatoma-derived cells. Alterations in hepatocellular metabolic and pro-oncogenic signaling pathways were assessed by immunoblotting.
We observed a positive correlation between the TAOK1 mRNA abundance in human liver biopsies and key hallmarks of NAFLD (i.e., hepatic steatosis, inflammation, and ballooning). Furthermore, we found that TAOK1 protein fully colocalized with intracellular lipid droplets in human and mouse hepatocytes. The silencing of TAOK1 alleviated lipotoxicity in cultured human hepatocytes by accelerating lipid catabolism (mitochondrial β-oxidation and triacylglycerol secretion), suppressing lipid anabolism (fatty acid influx and lipogenesis), and mitigating oxidative/endoplasmic reticulum stress, and the opposite changes were detected in TAOK1-overexpressing cells. We also found decreased proliferative, migratory, and invasive capacity, as well as lower epithelial-mesenchymal transition in TAOK1-deficient human hepatoma-derived cells. Mechanistic studies revealed that TAOK1 knockdown inhibited ERK and JNK activation and repressed acetyl-CoA carboxylase (ACC) protein abundance in human hepatocytes.
Together, we provide the first experimental evidence supporting the role of hepatic lipid droplet-decorating kinase TAOK1 in NAFLD development through mediating fatty acid partitioning between anabolic and catabolic pathways, regulating oxidative/endoplasmic reticulum stress, and modulating metabolic and pro-oncogenic signaling.
非酒精性脂肪性肝病(NAFLD)已成为全球导致慢性肝病的主要原因,影响了约四分之一的成年人口。NASH 是一种严重的非酒精性脂肪性肝病亚型,除了肝脂肪变性外,还意味着肝炎症和肝细胞气球样变。鉴于 NAFLD 的患病率呈指数级增长,我们必须更好地了解其分子发病机制。本研究旨在研究 STE20 型激酶 TAOK1(一种与肝细胞脂滴相关的蛋白)在调节肝脂毒性和 NAFLD 发病机制中的潜在作用。
在 62 名参与者的队列中,评估了肝活检中 TAOK1mRNA 表达与 NAFLD 严重程度的相关性。应用免疫荧光显微镜描述了 TAOK1 在人源和鼠源肝细胞中的亚细胞定位。通过功能测定、免疫荧光显微镜和比色分析,用小干扰 RNA 过表达或沉默 TAOK1 后,在永生化人肝细胞中研究了代谢重编程和氧化/内质网应激。在 TAOK1 缺陷的人肝癌衍生细胞中研究了迁移、侵袭和上皮-间充质转化。通过免疫印迹评估了肝细胞代谢和促癌信号通路的改变。
我们观察到人类肝活检中 TAOK1mRNA 的丰度与 NAFLD 的关键特征(即肝脂肪变性、炎症和气球样变)呈正相关。此外,我们发现 TAOK1 蛋白与人源和鼠源肝细胞中的细胞内脂滴完全共定位。TAOK1 沉默通过加速脂质分解代谢(线粒体β-氧化和三酰甘油分泌)、抑制脂质合成代谢(脂肪酸摄取和脂肪生成)以及减轻氧化/内质网应激,减轻了培养的人肝细胞的脂毒性,而在 TAOK1 过表达的细胞中则检测到相反的变化。我们还发现 TAOK1 缺陷的人肝癌衍生细胞增殖、迁移和侵袭能力降低,上皮-间充质转化水平降低。机制研究表明,TAOK1 沉默抑制了人肝细胞中 ERK 和 JNK 的激活,并抑制了乙酰辅酶 A 羧化酶(ACC)蛋白的丰度。
总之,我们提供了首个实验证据,支持肝脂滴修饰激酶 TAOK1 通过调节脂肪生成和分解代谢途径之间的脂肪酸分配、调节氧化/内质网应激以及调节代谢和促癌信号来参与 NAFLD 发生发展。