title: MicroRNA profiles in a monkey testicular injury model induced by testicular hyperthermia.
abstract: To characterize microRNAs (miRNAs) involved in testicular toxicity in cynomolgus monkeys, miRNA profiles were investigated using next-generation sequencing (NGS), microarray and reverse transcription-quantitative real-time-PCR (RT-qPCR) methods. First, to identify organ-specific miRNAs, we compared the expression levels of miRNAs in the testes to those in representative organs (liver, heart, kidney, lung, spleen and small intestine) obtained from naïve mature male and female monkeys (n = 2/sex) using NGS analysis. Consequently, miR-34c-5p, miR-202-5p, miR-449a and miR-508-3p were identified to be testicular-specific miRNAs in cynomolgus monkeys. Next, we investigated miRNA profiles after testicular-hyperthermia (TH) treatment to determine which miRNAs are involved in testicular injury. In this experiment, mature male monkeys were divided into groups with or without TH-treatment (n = 3/group) by immersion of the testes in a water bath at 43 °C for 30 min for 5 consecutive days. As a result, TH treatment induced testicular injury in all animals, which was characterized by decreased numbers of spermatocytes and spermatids. In a microarray analysis of the testis, 11 up-regulated (>2.0 fold) and 13 down-regulated (<0.5 fold) miRNAs were detected compared with those in the control animals. Interestingly, down-regulated miRNAs included two testicular-specific miRNAs, miR-34c-5p and miR-449a, indicating their potential use as biomarkers for testicular toxicity. Furthermore, RT-qPCR analysis revealed decreased expression levels of testicular miR-34b-5p and miR-34c-5p, which are enriched in meiotic cells, reflecting the decrease in pachytene spermatocytes and spermatids after TH treatment. These results provide valuable insights into the mechanism of testicular toxicity and potential translational biomarkers for testicular toxicity. Copyright © 2016 The Authors. Journal of Applied Toxicology published by John Wiley & Sons Ltd.
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd, 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo, Japan. sakurai.ken.t6@daiichisankyo.co.jp.
authors.forename: Ken
authors.initials: K
authors.lastname: Sakurai
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd, 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Mikamoto
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd, 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Shirai
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd, 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Iguchi
authors.affiliation: Translational Medicine & Clinical Pharmacology Department, Daiichi Sankyo Co., Ltd, 1-2-58 Hiromachi, Shinagawa-ku, Tokyo, Japan.
authors.forename: Kazumi
authors.initials: K
authors.lastname: Ito
authors.affiliation: Medicinal Safety Research Laboratories, Daiichi Sankyo Co., Ltd, 1-16-13 Kita-Kasai, Edogawa-ku, Tokyo, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Takasaki
authors.affiliation: Translational Medicine & Clinical Pharmacology, Daiichi Sankyo Pharma Development, Daiichi Sankyo, Inc., 399 Thornall Street, Edison, NJ, USA.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Mori
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D006358:Hot Temperature; D008252:Macaca fascicularis; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D009928:Organ Specificity; D013076:Sperm Count; D013087:Spermatids / Q000166:cytology / Q000187:drug effects; D013090:Spermatocytes / Q000166:cytology / Q000187:drug effects; D013737:Testis / Q000378:metabolism* / Q000473:pathology*; D013739:Testosterone / Q000097:blood; D059467:Transcriptome
chemical_list: D015415:Biomarkers; D035683:MicroRNAs; D013739:Testosterone
keywords: biomarker; cynomolgus monkey; microRNA; next-generation sequencing; testicular toxicity