Proteome-wide analyses of human hepatocytes during differentiation and dedifferentiation.
Proteome-wide analyses of human hepatocytes during differentiation and dedifferentiation.
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DOI：
10.1002/hep.26414
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发表时间：
2013-08
期刊：
影响因子：
13.5
通讯作者：
Hanley, Neil A.
中科院分区：
文献类型：
--
作者：
Rowe, Cliff;Gerrard, Dave T.;Jenkins, Roz;Berry, Andrew;Durkin, Kesta;Sundstrom, Lars;Goldring, Chris E.;Park, B. Kevin;Kitteringham, Neil R.;Hanley, Karen Piper;Hanley, Neil A.
在临床前试验中未能预测肝毒性药物，因此必须开发具有稳定表型的培养物的更好的肝脏模型。干细胞衍生的模型提供了希望，分化的肝细胞样细胞目前被认为是“胎儿样”的成熟。然而，这种判断是基于有限的生物标志物或转录本，缺乏直接比较胎儿和成人肝细胞所需的蛋白质组数据集。在这里，我们定量比较了人胎肝、成人肝细胞和HepG 2细胞系的蛋白质组。此外，我们研究了蛋白质组的变化，在人胎儿和成人肝细胞培养时，在一个新的空气-液体界面格式相比，传统的淹没细胞外基质夹心培养。根据白蛋白和尿素分泌以及基于胰蛋白酶的细胞色素P450活性，成年肝细胞在2周内的任一培养模型中均存活。胎细胞在气液界面系统中的功能得到较好的维持。引人注目的是，蛋白质组在所有样品中定性相似，但分层聚类显示每个样品类型具有不同的定量特征。HepG 2细胞更接近胎儿比成人肝细胞。此外，聚类显示，在空气-液体界面培养的原代成年肝细胞保留了一个蛋白质组，比传统培养更接近于模仿他们的新鲜同行，获得肌成纤维细胞的功能。主成分分析扩展了这些发现，并确定了一个简单的蛋白质，包括细胞色素P450 2A 6，谷胱甘肽S转移酶P，和酒精脱氢酶作为肝细胞分化的专门指标。结论：我们的定量数据集是第一个直接比较多个人类肝细胞的数据集，定义了一个模型，用于增强培养中肝细胞蛋白质组的维持，并提供了一个新的蛋白质“工具包”，用于确定培养细胞中人类肝细胞的成熟度。（肝病学2013;58：799-809）
Failure to predict hepatotoxic drugs in preclinical testing makes it imperative to develop better liver models with a stable phenotype in culture. Stem cell-derived models offer promise, with differentiated hepatocyte-like cells currently considered to be “fetal-like” in their maturity. However, this judgment is based on limited biomarkers or transcripts and lacks the required proteomic datasets that directly compare fetal and adult hepatocytes. Here, we quantitatively compare the proteomes of human fetal liver, adult hepatocytes, and the HepG2 cell line. In addition, we investigate the proteome changes in human fetal and adult hepatocytes when cultured in a new air-liquid interface format compared to conventional submerged extracellular matrix sandwich culture. From albumin and urea secretion, and luciferase-based cytochrome P450 activity, adult hepatocytes were viable in either culture model over 2 weeks. The function of fetal cells was better maintained in the air-liquid interface system. Strikingly, the proteome was qualitatively similar across all samples but hierarchical clustering showed that each sample type had a distinct quantitative profile. HepG2 cells more closely resembled fetal than adult hepatocytes. Furthermore, clustering showed that primary adult hepatocytes cultured at the air-liquid interface retained a proteome that more closely mimicked their fresh counterparts than conventional culture, which acquired myofibroblast features. Principal component analysis extended these findings and identified a simple set of proteins, including cytochrome P450 2A6, glutathione S transferase P, and alcohol dehydrogenases as specialized indicators of hepatocyte differentiation. Conclusion: Our quantitative datasets are the first that directly compare multiple human liver cells, define a model for enhanced maintenance of the hepatocyte proteome in culture, and provide a new protein “toolkit” for determining human hepatocyte maturity in cultured cells. (Hepatology 2013;58:799–809)
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影响因子：
3.5
作者：
Pochareddy S;Edenberg HJ
通讯作者：
Edenberg HJ
DOI：
10.1074/mcp.m800258-mcp200
发表时间：
2009-03
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Pan C;Kumar C;Bohl S;Klingmueller U;Mann M
通讯作者：
Mann M
影响因子：
5.2
作者：
Hay, David C.;Zhao, Debiao;Cui, Wei
通讯作者：
Cui, Wei
DOI：
10.1124/jpet.300.2.355
发表时间：
2002-02-01
影响因子：
3.5
作者：
Hines, RN;McCarver, DG
通讯作者：
McCarver, DG
DOI：
10.1124/jpet.102.053124
发表时间：
2003-10-01
影响因子：
3.5
作者：
Johnsrud, EK;Koukouritaki, SB;McCarver, DG
通讯作者：
McCarver, DG