title: Detection of cell-free, liver-specific mRNAs in peripheral blood from rats with hepatotoxicity: a potential toxicological biomarker for safety evaluation.
abstract: To verify the concept that cell-free organ/tissue-specific mRNAs leaking from drug-damaged organs/tissues into peripheral blood could be toxicological biomarkers for identification of the target organs of drug toxicity, we attempted to detect liver-specific mRNAs in peripheral blood from rats with chemical-induced hepatotoxicity. We selected alpha(1)-microglobulin/bikunin precursor (Ambp) and albumin mRNAs as tentative liver-specific biomarkers and successfully detected them by reverse transcription (RT)-PCR in peripheral blood 24 h after D-galactosamine HCl (D-gal) or acetaminophen administration. Moreover, albumin mRNA was detected 2 h after D-gal administration, although plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were still unchanged. On the other hand, in peripheral blood from rat with bupivacaine HCl-induced skeletal muscle damage, neither Ambp nor albumin mRNA was detectable while plasma creatine kinase, ALT, and AST levels prominently increased 2 or 12 h after dosing. Furthermore, Ambp mRNA was also detectable in filtered plasma from rats with liver damage, indicating that cell-free Ambp mRNA can be present in peripheral blood. In conclusion, cell-free, liver-specific Ambp, and albumin mRNAs were detectable in peripheral blood from rats with chemical-induced liver damage. It is believed that the detection of cell-free organ/tissue-specific mRNA in peripheral blood is a promising approach in the survey of toxicological biomarkers.
authors.affiliation: Development Research Center, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, Yodogawa-ku, Osaka 532-8686, Japan. Miyamoto_Makoto@takeda.co.jp
authors.forename: Makoto
authors.initials: M
authors.lastname: Miyamoto
authors.forename: Mariko
authors.initials: M
authors.lastname: Yanai
authors.forename: Shingo
authors.initials: S
authors.lastname: Ookubo
authors.forename: Naoko
authors.initials: N
authors.lastname: Awasaki
authors.forename: Kenji
authors.initials: K
authors.lastname: Takami
authors.forename: Ryoetsu
authors.initials: R
authors.lastname: Imai
mesh_terms: D000418:Albumins / Q000235:genetics; D000510:Alpha-Globulins; D000818:Animals; D001483:Base Sequence; D015415:Biomarkers / Q000097:blood*; D002474:Cell-Free System; D056486:Chemical and Drug Induced Liver Injury / Q000097:blood / Q000235:genetics*; D017931:DNA Primers; D005260:Female; D008297:Male; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000097:blood*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D000418:Albumins; D000510:Alpha-Globulins; D015415:Biomarkers; D017931:DNA Primers; D011506:Proteins; D012333:RNA, Messenger; C016001:alpha-1-microglobulin