Detection of cell-free, liver-specific mRNAs in peripheral blood from rats with hepatotoxicity: a potential toxicological biomarker for safety evaluation.
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.
Miyamoto
Makoto
M
Development Research Center, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, Yodogawa-ku, Osaka 532-8686, Japan. Miyamoto_Makoto@takeda.co.jp
Yanai
Mariko
M
Ookubo
Shingo
S
Awasaki
Naoko
N
Takami
Kenji
K
Imai
Ryoetsu
R
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*
D000418:Albumins; D000510:Alpha-Globulins; D015415:Biomarkers; D017931:DNA Primers; D011506:Proteins; D012333:RNA, Messenger; C016001:alpha-1-microglobulin