1. Circulating miRNAs, isomiRs and small RNA clusters in human plasma and breast milk
- Author
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Yaris Sarria, Maria Riera Piqué-Borràs, Alfredo Mayor, Ramon Carreras, Kilian Vellvé, Mariona Bustamante, Juan R. González, Dietmar Fernández-Orth, Lorena Pantano, Carles Hernandez-Ferrer, Xavier Estivill, Silvia Agramunt, and Mercedes Rubio
- Subjects
Male ,0301 basic medicine ,Small RNA ,Micro RNAs ,Physiology ,Molecular biology ,lcsh:Medicine ,Biochemistry ,Sequencing techniques ,Medicine and Health Sciences ,Small nucleolar RNAs ,Small nucleolar RNA ,lcsh:Science ,Breast Milk ,Multidisciplinary ,RNA sequencing ,Healthy Volunteers ,Body Fluids ,Nucleic acids ,Milk ,Blood ,Transfer RNA ,Female ,Anatomy ,Research Article ,Adult ,Breast milk ,Llet materna -- Investigació ,Biology ,Blood Plasma ,Beverages ,Young Adult ,03 medical and health sciences ,Extraction techniques ,microRNA ,Genetics ,Humans ,Non-coding RNA ,Nutrition ,Biology and life sciences ,Milk, Human ,lcsh:R ,RNA ,RNA extraction ,Gene regulation ,Diet ,Research and analysis methods ,MicroRNAs ,Molecular biology techniques ,030104 developmental biology ,Human plasma ,Llet materna ,lcsh:Q ,Gene expression ,Small nuclear RNA - Abstract
Circulating small RNAs, including miRNAs but also isomiRs and other RNA species, have the potential to be used as non-invasive biomarkers for communicable and non-communicable diseases. This study aims to characterize and compare small RNA profiles in human biofluids. For this purpose, RNA was extracted from plasma and breast milk samples from 15 healthy postpartum mothers. Small RNA libraries were prepared with the NEBNext(R) small RNA library preparation kit and sequenced in an Illumina HiSeq2000 platform. miRNAs, isomiRs and clusters of small RNAs were annotated using seqBuster/seqCluster framework in 5 plasma and 10 milk samples that passed the initial quality control. The RNA yield was 81 ng/mL [standard deviation (SD): 41] and 3985 ng/mL (SD: 3767) for plasma and breast milk, respectively. Mean number of good quality reads was 4.04 million (M) (40.01% of the reads) in plasma and 12.5M (89.6%) in breast milk. One thousand one hundred eighty two miRNAs, 12,084 isomiRs and 1,053 small RNA clusters that included piwi-interfering RNAs (piRNAs), tRNAs, small nucleolar RNAs (snoRNA) and small nuclear RNAs (snRNAs) were detected. Samples grouped by biofluid, with 308 miRNAs, 1,790 isomiRs and 778 small RNA clusters differentially detected. In summary, plasma and milk showed a different small RNA profile. In both, miRNAs, piRNAs, tRNAs, snRNAs, and snoRNAs were identified, confirming the presence of non-miRNA species in plasma, and describing them for the first time in milk.
- Published
- 2018