Modelling non-alcoholic fatty liver disease in human hepatocyte-like cells.
Modelling non-alcoholic fatty liver disease in human hepatocyte-like cells.
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DOI：
10.1098/rstb.2017.0362
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发表时间：
2018-07-05
期刊：
影响因子：
--
通讯作者：
Drake AJ
中科院分区：
文献类型：
--
作者：
Lyall MJ;Cartier J;Thomson JP;Cameron K;Meseguer-Ripolles J;O'Duibhir E;Szkolnicka D;Villarin BL;Wang Y;Blanco GR;Dunn WB;Meehan RR;Hay DC;Drake AJ
非酒精性脂肪性肝病（NAFLD）是发达国家最常见的肝病原因。体外NAFLD模型将允许机制研究并实现高通量治疗筛选。虽然肝癌来源的细胞系是一种方便的可再生资源，但它们的基因组、表观基因组和功能改变意味着它们在NAFLD建模中的效用尚不清楚。此外，表观遗传标记5-羟甲基胞嘧啶（5 hmC），细胞谱系标识符，在细胞培养过程中迅速丢失，同时表达十-十一易位（泰特）甲基胞嘧啶双加氧酶，限制了有意义的表观遗传分析。来源于人胚胎干细胞的肝细胞样细胞（HLC）可以为肝脏研究提供一种非肿瘤性、可再生的模型。在这里，我们使用暴露于乳酸、丙酮酸和辛酸（LPO）的HLC开发了一种NAFLD模型，这些HLC具有肝脏功能的所有标志，包括5 hmC特征。我们将HLCs暴露于LPO 48 h以诱导脂质积聚。我们使用RNA-seq表征转录组，使用超高效液相色谱-质谱法表征代谢组，使用5-羟甲基化DNA免疫沉淀（hmeDIP）测序表征表观基因组。LPO暴露诱导HLCs中的NAFLD表型，其转录和代谢组学失调与人NAFLD中存在的那些一致。HLC维持泰特酶的表达，并具有肝样表观基因组。脂质过氧化氢诱导的5 hmC富集在脂质合成和转运基因。用LPO处理的HLC重现了在NAFLD中观察到的转录和代谢失调，并且另外保留了泰特表达和5 hmC。这种NAFLD的体外模型将有助于未来的机制和治疗研究。这篇文章是“设计师人体组织：来到你附近的实验室”主题的一部分。
Non-alcoholic fatty liver disease (NAFLD) is the most common cause of liver disease in developed countries. An in vitro NAFLD model would permit mechanistic studies and enable high-throughput therapeutic screening. While hepatic cancer-derived cell lines are a convenient, renewable resource, their genomic, epigenomic and functional alterations mean their utility in NAFLD modelling is unclear. Additionally, the epigenetic mark 5-hydroxymethylcytosine (5hmC), a cell lineage identifier, is rapidly lost during cell culture, alongside expression of the Ten-eleven-translocation (TET) methylcytosine dioxygenase enzymes, restricting meaningful epigenetic analysis. Hepatocyte-like cells (HLCs) derived from human embryonic stem cells can provide a non-neoplastic, renewable model for liver research. Here, we have developed a model of NAFLD using HLCs exposed to lactate, pyruvate and octanoic acid (LPO) that bear all the hallmarks, including 5hmC profiles, of liver functionality. We exposed HLCs to LPO for 48 h to induce lipid accumulation. We characterized the transcriptome using RNA-seq, the metabolome using ultra-performance liquid chromatography-mass spectrometry and the epigenome using 5-hydroxymethylation DNA immunoprecipitation (hmeDIP) sequencing. LPO exposure induced an NAFLD phenotype in HLCs with transcriptional and metabolomic dysregulation consistent with those present in human NAFLD. HLCs maintain expression of the TET enzymes and have a liver-like epigenome. LPO exposure-induced 5hmC enrichment at lipid synthesis and transport genes. HLCs treated with LPO recapitulate the transcriptional and metabolic dysregulation seen in NAFLD and additionally retain TET expression and 5hmC. This in vitro model of NAFLD will be useful for future mechanistic and therapeutic studies. This article is part of the theme issue ‘Designer human tissue: coming to a lab near you’.
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影响因子：
12.3
作者：
Nestor CE;Ottaviano R;Reinhardt D;Cruickshanks HA;Mjoseng HK;McPherson RC;Lentini A;Thomson JP;Dunican DS;Pennings S;Anderton SM;Benson M;Meehan RR
通讯作者：
Meehan RR
影响因子：
29
作者：
Pavlova NN;Thompson CB
通讯作者：
Thompson CB
影响因子：
6.1
作者：
Lucendo-Villarin B;Filis P;Swortwood MJ;Huestis MA;Meseguer-Ripolles J;Cameron K;Iredale JP;O'Shaughnessy PJ;Fowler PA;Hay DC
通讯作者：
Hay DC
影响因子：
6
作者：
Medine, Claire N.;Lucendo-Villarin, Baltasar;Hay, David C.
通讯作者：
Hay, David C.
DOI：
10.1073/pnas.1415301111
发表时间：
2015-06-02
影响因子：
11.1
作者：
Bestor, Timothy H.;Edwards, John R.;Boulard, Mathieu
通讯作者：
Boulard, Mathieu