assessment. Recently, using EV-associated miRNAs or circulating free miRNAs in serum as biomarkers has made it possible to diagnose thirteen types of cancer with an accuracy of 90% or more (Ogata-Kawata et al. 2014; Shimomura et al. 2016; Yokoi et al. 2018; Yao et al. 2019; Asano et al. 2019; Shiino et al. 2019; Usuba et al. 2019; Asakura et al. 2020). The development of next-generation toxicity tests using miRNA as a biomarker is therefore expected.
Our studies identified forty-two novel miRNAs—such as miR-122 and miR-192—as candidate liver damage biomarkers. Using these novel biomarkers, it may be possible to elucidate the mechanism of hepatotoxicity caused by the administration of drugs or chemical substances other than carbon tetrachloride. Since EVs in blood are secreted by a wide variety of cells, it is necessary to clarify the origin of the EV-associated miRNAs. In addition to their utility as markers of hepatotoxicity, EV-associated miRNAs are expected to be valuable as biomarkers of toxicity that targets other organs, such as the kidney, the lung, and the heart.
Although this study analyzed the effects of carbon tetrachloride administration at a single time point, twenty-four hours, evaluating the time courses of biomarkers in response to the repeated administration of carbon tetrachloride may be applicable if we want to shorten chronic toxicity tests and long-term carcinogenicity tests.
It may also be important to elucidate the function of the identified EV-associated miRNAs induced by hepatotoxicity in vivo.
Acknowledgments
The authors thank N. Moriyama, T. Momiyama, E. Tachihara, M. Uchiyama, and H. Aihara for excellent technical assistance. The images of mice, cows and cultured cells are from TogoTV (©2016 DBCLS TogoTV/CC-BY-4.0).
Competing Interests
The authors declare no competing interests.
Funding
This work was supported in part by Research on the Regulatory Science of Pharmaceuticals and Medical Devices (20mk0101163j0202), Research on the Development of New Drugs (20ak0101093j003) from the Japan Agency for Medical Research and Development, the Health Sciences Research Grants from the Ministry of Health, Labor, and Welfare, Japan, (H30-KAGAKU-IPPAN-002, 21KD1001), and JSPS KAKENHI (18K19315) to R.O.; Grant-in-Aid from the Research Program on Hepatitis from Japan Agency for Medical Research and Development (AMED: 16fk0310512h0005, 17fk0310101h0001 and 18fk0310101h0002) and Grants-in -Aid from a Project for Cancer research and Therapeutic Evolution (P-CREATE) to T. O.; JSPS KAKENHI (18K07053) to Y. H.
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26 De