title: A toxicogenomic approach revealed hepatic gene expression changes mechanistically linked to drug-induced hemolytic anemia.
abstract: A variety of pharmaceutical compounds causes hemolytic anemia as a significant adverse effect and this toxicity restricts the clinical utility of these drugs. In this study, we applied microarray technology to investigate hepatic gene expression changes associated with drug-induced hemolytic anemia and to identify potential biomarker genes for this hematotoxicity. We treated female Sprague-Dawley rats with two hemolytic anemia-inducing compounds: phenylhydrazine and phenacetin. Hepatic gene expression profiles were obtained using a whole-genome oligonucleotide microarray with pooled RNA samples from individual rats within each dose group and analyzed in comparison with hepatic histopathology, hematology, and blood chemistry data. We identified a small subset of genes that were commonly deregulated in all the severe hemolytic conditions, some of which were considered to be involved in hepatic events characteristic of hemolytic anemia, such as hemoglobin biosynthesis, heme metabolism, and phagocytosis. Among them, we selected six upregulated genes as putative biomarkers, and their expression changes from microarray measurements were confirmed by quantitative real-time PCR using RNAs from individual animals. They were Alas2, beta-glo, Eraf, Hmox1, Lgals3, and Rhced. Expression patterns of all these genes showed high negative and positive correlation against erythrocyte counts and total bilirubin levels in circulation, respectively, suggesting that these genes may be the potential biomarkers for hemolytic anemia. These findings indicate that drug-induced hemolytic anemia may be detected based on hepatic changes in the expression of a subset of genes that are mechanistically linked to the hematotoxicity.
authors.affiliation: Discovery Technologies 1, Discovery Research Laboratories, Shionogi and Co Ltd, 12-4, Sagisu 5-chome, Fukushima-ku, Osaka 553-0002, Japan. masatomo.rokushima@shionogi.co.jp
authors.forename: Masatomo
authors.initials: M
authors.lastname: Rokushima
authors.forename: Kazuo
authors.initials: K
authors.lastname: Omi
authors.forename: Akiko
authors.initials: A
authors.lastname: Araki
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Kyokawa
authors.forename: Naoko
authors.initials: N
authors.lastname: Furukawa
authors.forename: Fumio
authors.initials: F
authors.lastname: Itoh
authors.forename: Kae
authors.initials: K
authors.lastname: Imura
authors.forename: Kumiko
authors.initials: K
authors.lastname: Takeuchi
authors.forename: Manabu
authors.initials: M
authors.lastname: Okada
authors.forename: Ikuo
authors.initials: I
authors.lastname: Kato
authors.forename: Jun
authors.initials: J
authors.lastname: Ishizaki
mesh_terms: D000743:Anemia, Hemolytic / Q000097:blood / Q000139:chemically induced* / Q000235:genetics; D000818:Animals; D001663:Bilirubin / Q000097:blood; D015536:Down-Regulation; D064420:Drug-Related Side Effects and Adverse Reactions; D004906:Erythrocyte Count; D004912:Erythrocytes / Q000166:cytology / Q000187:drug effects; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D005819:Genetic Markers / Q000235:genetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D010615:Phenacetin / Q000009:adverse effects; D010659:Phenylhydrazines / Q000009:adverse effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
chemical_list: D005819:Genetic Markers; D010659:Phenylhydrazines; C030299:phenylhydrazine; D010615:Phenacetin; D001663:Bilirubin