Department of Statistics, TU Dortmund University, Dortmund, Germany.
Arch Toxicol. 2014 Dec;88(12):2261-87. doi: 10.1007/s00204-014-1400-x. Epub 2014 Nov 16.
A long-term goal of numerous research projects is to identify biomarkers for in vitro systems predicting toxicity in vivo. Often, transcriptomics data are used to identify candidates for further evaluation. However, a systematic directory summarizing key features of chemically influenced genes in human hepatocytes is not yet available. To bridge this gap, we used the Open TG-GATES database with Affymetrix files of cultivated human hepatocytes incubated with chemicals, further sets of gene array data with hepatocytes from human donors generated in this study, and publicly available genome-wide datasets of human liver tissue from patients with non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular cancer (HCC). After a curation procedure, expression data of 143 chemicals were included into a comprehensive biostatistical analysis. The results are summarized in the publicly available toxicotranscriptomics directory ( http://wiki.toxbank.net/toxicogenomics-map/ ) which provides information for all genes whether they are up- or downregulated by chemicals and, if yes, by which compounds. The directory also informs about the following key features of chemically influenced genes: (1) Stereotypical stress response. When chemicals induce strong expression alterations, this usually includes a complex but highly reproducible pattern named 'stereotypical response.' On the other hand, more specific expression responses exist that are induced only by individual compounds or small numbers of compounds. The directory differentiates if the gene is part of the stereotypical stress response or if it represents a more specific reaction. (2) Liver disease-associated genes. Approximately 20 % of the genes influenced by chemicals are up- or downregulated, also in liver disease. Liver disease genes deregulated in cirrhosis, HCC, and NASH that overlap with genes of the aforementioned stereotypical chemical stress response include CYP3A7, normally expressed in fetal liver; the phase II metabolizing enzyme SULT1C2; ALDH8A1, known to generate the ligand of RXR, one of the master regulators of gene expression in the liver; and several genes involved in normal liver functions: CPS1, PCK1, SLC2A2, CYP8B1, CYP4A11, ABCA8, and ADH4. (3) Unstable baseline genes. The process of isolating and the cultivation of hepatocytes was sufficient to induce some stress leading to alterations in the expression of genes, the so-called unstable baseline genes. (4) Biological function. Although more than 2,000 genes are transcriptionally influenced by chemicals, they can be assigned to a relatively small group of biological functions, including energy and lipid metabolism, inflammation and immune response, protein modification, endogenous and xenobiotic metabolism, cytoskeletal organization, stress response, and DNA repair. In conclusion, the introduced toxicotranscriptomics directory offers a basis for a rationale choice of candidate genes for biomarker evaluation studies and represents an easy to use source of background information on chemically influenced genes.
长期以来,许多研究项目的目标是确定用于预测体内毒性的体外系统的生物标志物。通常,转录组学数据用于识别进一步评估的候选者。然而,目前还没有一个系统目录来总结化学物质影响人类肝细胞中基因的关键特征。为了弥补这一空白,我们使用了 Open TG-GATES 数据库和 Affymetrix 文件,其中包含用化学物质孵育的培养人肝细胞,以及本研究中生成的来自人类供体的肝细胞的进一步基因阵列数据集,以及来自非酒精性脂肪性肝炎 (NASH)、肝硬化和肝细胞癌 (HCC) 患者的公开可用的人类肝组织全基因组数据集。经过整理程序,将 143 种化学物质的表达数据纳入综合生物统计学分析。结果总结在公开的毒代转录组学目录(http://wiki.toxbank.net/toxicogenomics-map/)中,该目录提供了所有基因的信息,无论它们是否被化学物质上调或下调,如果是,是由哪些化合物引起的。该目录还提供了受化学物质影响的基因的以下关键特征的信息:(1)刻板的应激反应。当化学物质引起强烈的表达改变时,这通常包括一种复杂但高度可重复的模式,称为“刻板反应”。另一方面,存在更具体的表达反应,仅由单个化合物或少数化合物诱导。目录区分基因是否属于刻板的应激反应,或者它是否代表更具体的反应。(2)与肝脏疾病相关的基因。大约 20%受化学物质影响的基因在肝脏疾病中也会上调或下调。在肝硬化、肝癌和 NASH 中失调的与上述典型化学应激反应相关的肝脏疾病基因包括 CYP3A7,它在胎儿肝脏中表达;相 II 代谢酶 SULT1C2;ALDH8A1,已知产生 RXR 的配体,RXR 是肝脏中基因表达的主要调节剂之一;以及几个参与正常肝脏功能的基因:CPS1、PCK1、SLC2A2、CYP8B1、CYP4A11、ABCA8 和 ADH4。(3)不稳定的基线基因。分离和培养肝细胞的过程足以引起一些应激,导致基因表达的改变,即所谓的不稳定基线基因。(4)生物学功能。尽管有 2000 多个基因受到化学物质的转录影响,但它们可以归为相对较小的生物学功能组,包括能量和脂质代谢、炎症和免疫反应、蛋白质修饰、内源性和外源性代谢、细胞骨架组织、应激反应和 DNA 修复。总之,引入的毒代转录组学目录为候选基因用于生物标志物评估研究提供了一个合理的选择基础,并代表了一个易于使用的化学物质影响基因背景信息来源。