title: Pyrene did not induce gene mutation in red blood cell Pig-a assay and PIGRET assay in rats.
abstract: A new in vivo gene mutation assay has been developed based on the phosphatidylinositol glycan anchor biosynthesis, Class A gene (Pig-a in rodents) as an endogenous reporter. Although a large number of chemicals have been evaluated in the rat Pig-a assay in 28-day repeat dose regimens, there was limited reporting of rat Pig-a assay after a single dose. A collaborative study by the Mammalian Mutagenicity Study group, which is a subgroup of the Japanese Environmental Mutagen Society, was conducted to verify the usefulness of the rat Pig-a assay after a single dose as a short-term genotoxicity test. As a part of this collaborative study, the in vivo mutagenicity of a single dose of pyrene (Pyr) was investigated in the red blood cell (RBC Pig-a assay) and in reticulocytes (PIGRET) of rats. Eight-week old male rats were orally dosed with Pyr at 500, 1000, and 2000 mg/kg or ethylnitrosourea (ENU) at 10 and 40 mg/kg as a positive control. The animals in each group were examined for Pig-a mutant frequencies (MF) except for animals in the 2000mg/kg group because of mortality or severe toxicity. The Pig-a MF in RBCs and reticulocytes, as CD59 negative cells, were evaluated once a week for 4 weeks after the dosing. With a single exposure to ENU, the Pig-a MF in both RBCs and reticulocytes increased in a time- and dose-dependent manner. In contrast, no statistically significant effect was observed in rats dosed with Pyr at 500 and 1000 mg/kg. Therefore, Pyr was concluded to be negative in the RBC Pig-a assay and the PIGRET assay after a single oral administration in rats. The result was consistent with previously reported Pig-a assays with repeat dose regimens.
authors.affiliation: Drug Safety Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, 251-8555, Japan.
authors.forename: Ikuma
authors.initials: I
authors.lastname: Yoshida
authors.affiliation: Drug Safety Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, 251-8555, Japan.
authors.forename: Akemi
authors.initials: A
authors.lastname: Matsumoto
authors.affiliation: Drug Safety Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, 251-8555, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Sakai
authors.affiliation: Drug Safety Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, 251-8555, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Harada
authors.affiliation: Drug Safety Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, 251-8555, Japan. Electronic address: tsuneo.hashizume@takeda.com.
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Hashizume
mesh_terms: D000818:Animals; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity*; D009154:Mutation; D011721:Pyrenes / Q000633:toxicity*; D051381:Rats; D012156:Reticulocytes / Q000187:drug effects*
chemical_list: D008565:Membrane Proteins; D009153:Mutagens; D011721:Pyrenes; C080086:phosphatidylinositol glycan-class A protein; C030984:pyrene
keywords: Gene mutation; Pig-a gene; Pyrene; Red blood cells; Reticulocytes