Human plasma and serum extracellular small RNA reference profiles and their clinical utility

Proceedings of the National Academy of Sciences of the United States of America
Klaas E A MaxThomas Tuschl

Abstract

Circulating extracellular RNAs (exRNAs) have the potential to serve as biomarkers for a wide range of medical conditions. However, limitations in existing exRNA isolation methods and a lack of knowledge on parameters affecting exRNA variability in human samples may hinder their successful discovery and clinical implementation. Using combinations of denaturants, reducing agents, proteolysis, and revised organic extraction, we developed an automated, high-throughput approach for recovery of exRNAs and exDNA from the same biofluid sample. We applied this method to characterize exRNAs from 312 plasma and serum samples collected from 13 healthy volunteers at 12 time points over a 2-month period. Small RNA cDNA library sequencing identified nearly twofold increased epithelial-, muscle-, and neuroendocrine-cell-specific miRNAs in females, while fasting and hormonal cycle showed little effect. External standardization helped to detect quantitative differences in erythrocyte and platelet-specific miRNA contributions and in miRNA concentrations between biofluids. It also helped to identify a study participant with a unique exRNA phenotype featuring a miRNA signature of up to 20-fold elevated endocrine-cell-specific miRNAs and twofold ele...Continue Reading

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Methods Mentioned

BETA
chromosomal aberrations
cDNA library
nucleic acid purification
blood draw
dissect
PCR
RNAseq
exRNA-seq

Software Mentioned

pheatmap
RSDAP
exRNA atlas
DESeq2
heatmap
GraphPad Prism
DEA
ReportingTools
stat
Illumina RNA - sequencing data analysis platform ( RSDAP )

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