title: Comparing next-generation sequencing and microarray technologies in a toxicological study of the effects of aristolochic acid on rat kidneys.
abstract: RNA-Seq has been increasingly used for the quantification and characterization of transcriptomes. The ongoing development of the technology promises the more accurate measurement of gene expression. However, its benefits over widely accepted microarray technologies have not been adequately assessed, especially in toxicogenomics studies. The goal of this study is to enhance the scientific community's understanding of the advantages and challenges of RNA-Seq in the quantification of gene expression by comparing analysis results from RNA-Seq and microarray data on a toxicogenomics study. A typical toxicogenomics study design was used to compare the performance of an RNA-Seq approach (Illumina Genome Analyzer II) to a microarray-based approach (Affymetrix Rat Genome 230 2.0 arrays) for detecting differentially expressed genes (DEGs) in the kidneys of rats treated with aristolochic acid (AA), a carcinogenic and nephrotoxic chemical most notably used for weight loss. We studied the comparability of the RNA-Seq and microarray data in terms of absolute gene expression, gene expression patterns, differentially expressed genes, and biological interpretation. We found that RNA-Seq was more sensitive in detecting genes with low expression levels, while similar gene expression patterns were observed for both platforms. Moreover, although the overlap of the DEGs was only 40-50%, the biological interpretation was largely consistent between the RNA-Seq and microarray data. RNA-Seq maintained a consistent biological interpretation with time-tested microarray platforms while generating more sensitive results. However, there is clearly a need for future investigations to better understand the advantages and limitations of RNA-Seq in toxicogenomics studies and environmental health research.
authors.affiliation: ICF International at FDA's National Center for Toxicological Research, 3900 NCTR Road, Jefferson, Arkansas 72079, USA. zhenqiang.su@fda.hhs.gov
authors.forename: Zhenqiang
authors.initials: Z
authors.lastname: Su
authors.forename: Zhiguang
authors.initials: Z
authors.lastname: Li
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.forename: Quan-Zhen
authors.initials: QZ
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Fang
authors.forename: Don
authors.initials: D
authors.lastname: Ding
authors.forename: Weigong
authors.initials: W
authors.lastname: Ge
authors.forename: Baitang
authors.initials: B
authors.lastname: Ning
authors.forename: Huixiao
authors.initials: H
authors.lastname: Hong
authors.forename: Roger G
authors.initials: RG
authors.lastname: Perkins
authors.forename: Weida
authors.initials: W
authors.lastname: Tong
authors.forename: Leming
authors.initials: L
authors.lastname: Shi
mesh_terms: D000818:Animals; D034341:Aristolochic Acids / Q000633:toxicity*; D015197:Carcinogenicity Tests / Q000379:methods; D002273:Carcinogens / Q000633:toxicity*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D051381:Rats; D017423:Sequence Analysis, RNA / Q000379:methods*; D043922:Toxicogenetics / Q000379:methods
chemical_list: D034341:Aristolochic Acids; D002273:Carcinogens