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Published on: August 21, 2019
Analysis of gene expression for microminipig liver transcriptomes using parallel long-read technology and short-read
Chizuka Sakai1, Shunsuke Iwano1,2, Makiko Shimizu2
1Toxicology and Pharmacokinetics Laboratories, Pharmaceutical Research Laboratories, Toray Industries, Inc., Kamakura, Kanagawa, 248-8555, Japan.
Abstract:
The microminipig is one of the smallest minipigs that has emerged as a possible experimental animal model, because it shares many anatomical and/or physiological similarities with humans, including the coronary artery distribution in the heart, the digestive physiology, the kidney size and its structure, and so on. However, information on gene expression profiles, including those on drug-metabolizing phase I and II enzymes, in the microminipig is limited. Therefore, the aim of the present study was to identify transcripts in microminipig livers and to determine gene expression profiles. De novo assembly and expression analyses of microminipig transcripts were conducted with liver samples from three male and three female microminipigs using parallel long-read and short-read sequencing technologies. After unique sequences had been automatically aligned by assembling software, the mean contig length of 50843 transcripts was 707 bp. The expression profiles of cytochrome P450 (P450) 1A2, 2C, 2E1 and 3A genes in livers in microminipigs were similar to those in humans. Liver carboxylesterase (CES) precursor, liver CES-like, UDP-glucuronosyltransferase (UGT) 2C1-like, amine sulfotransferase (SULT)-like, N-acetyltransferases (NAT8) and glutathione S-transferase (GST) A2 genes, which are relatively unknown genes in pigs and/or humans, were expressed strongly. Furthermore, no significant gender differences were observed in the gene expression profiles of phase I enzymes, whereas UGT2B17, SULT1E1, SULT2A1, amine SULT-like, NAT8 and GSTT4 genes were different between males and females among phase II enzyme genes under the present sample conditions. These results provide a foundation for mechanistic studies and the use of microminipigs as model animals for drug development in the future. Copyright © 2016 John Wiley & Sons, Ltd.
Insights
Micropigs offer valuable insights into human drug metabolism. This study profiles liver gene expression, revealing similarities to humans and identifying key drug-metabolizing enzymes for future research.
Area of Science:
- Pharmacogenomics
- Animal Models
- Biotechnology
Background:
- Micropigs share anatomical and physiological similarities with humans, making them potential experimental models.
- Limited data exists on gene expression profiles, particularly for drug-metabolizing enzymes, in micropigs.
- Understanding these profiles is crucial for utilizing micropigs in preclinical drug development.
Purpose of the Study:
- To identify transcripts in micropig livers.
- To determine the gene expression profiles of drug-metabolizing enzymes in micropigs.
- To compare these profiles with human data and assess gender-specific differences.
Main Methods:
- Liver samples from six micropigs (three male, three female) were analyzed using parallel long-read and short-read sequencing.
- De novo assembly and expression analyses were performed on the obtained transcripts.
- Gene expression levels of phase I and phase II drug-metabolizing enzymes were quantified.
Main Results:
- A total of 50,843 transcripts were identified, with a mean contig length of 707 bp.
- Expression profiles of cytochrome P450 (P450) 1A2, 2C, 2E1, and 3A genes were similar to those in humans.
- Several phase II enzyme genes, including UDP-glucuronosyltransferase (UGT) and glutathione S-transferase (GST) variants, showed strong expression and some gender-specific differences.
Conclusions:
- Micropig liver gene expression profiles, especially for drug-metabolizing enzymes, provide a foundation for their use in drug development.
- The observed similarities to human gene expression support the micropig as a relevant preclinical model.
- Further research can leverage these findings for mechanistic studies and enhanced drug efficacy and safety evaluations.
