Landscape of circRNAs Across 11 Organs and 4 Ages in Fischer 344 Rats.

Circular RNAs (circRNAs) are a class of endogenous noncoding RNAs with a covalently closed loop. Aside from their recognized regulatory functions (e.g., sponging microRNAs to reduce their activity, and altering parental gene transcription by competing with the canonical splicing of pre-mRNA), expression of circRNAs is abundant, diverse, and conservative across species, rendering them as potential biomarker candidates. Consequently, the landscape of circRNAs has been studied for several species. Although the rat is one of the most important animal models for drug safety and toxicological research, few attempts have been made to understand the landscape of rat circRNAs. One noticeable challenge in analyzing circRNAs with next-generation sequencing (NGS) data is to find ways to use rapidly advancing bioinformatics approaches to improve accuracy while also reducing the number of resulting false positives that occur in circRNA identification with these new methods. Here, we applied two of the most advanced circRNA bioinformatics pipelines to provide a landscape of circRNAs in rats by analyzing an RNA-seq data set for 11 organs (adrenal gland, brain, heart, kidney, liver, lung, muscle, spleen, thymus, and testis or uterus) from Fischer 344 rats of both sexes in four age groups (juvenile, adolescence, adult, and aged). The circRNAs displayed organ-specific patterns and sex differences in most organs. Lowest numbers of circRNAs were seen in the liver and muscle, while highest numbers of circRNAs occurred in the brain, which correlated to gene expression patterns seen across those organs. Concordance of circRNAs between males and females was approximately 50% in nonsex organs, implying that some caution needs to be exercised when selecting specific circRNAs as biomarkers for both sexes. The number of common circRNAs between sexes increased with age for most organs except heart, spleen, and thymus. A dramatic drop in the number of circRNAs in kidney, thymus, and testis was observed in aged rats, suggesting that the regulatory function of circRNAs is age dependent. From the 1595 circRNAs identified with high confidence, only 6 appeared in all 9 of the nonsex organs in both sexes and four age groups. Forty-one and 48 circRNAs were identified in more than 5 nonsex organs in males and females, respectively, while close to 280 circRNAs were found in an organ for more than 2 age groups in both sexes. This study offers an overview of rat circRNAs, which contributes to the effort of identifying circRNAs as potential biomarkers for safety and risk assessment.