Detection of circulating extracelluar mRNAs by modified small-RNA-sequencing analysis
Akat KM, Lee YA, Hurley A, Morozov P, Max KEA, Brown M, Bogardus K, Sopeyin A, Hildner K, Diacovo TG, Neurath M, Borggrefe M, Tuschl T (2019)
Publication Type: Journal article
Publication year: 2019
Journal
Book Volume: 4
Journal Issue: 9
DOI: 10.1172/jci.insight.127317
Abstract
Extracellular mRNAs (ex-mRNAs) potentially supersede extracellular miRNAs (ex-miRNAs) and other RNA classes as biomarkers. We performed conventional small-RNA-sequencing (sRNA-seq) and sRNA-seq with T4 polynucleotide kinase (PNK) end treatment of total extracellular RNAs (exRNAs) isolated from serum and platelet-poor EDTA, acid citrate dextrose (ACD), and heparin plasma to study the effect on ex-mRNA capture. Compared with conventional sRNA-seq, PNK treatment increased the detection of informative ex-mRNAs reads up to 50-fold. The exRNA pool was dominated by RNA originating from hematopoietic cells and platelets, with additional contribution from the liver. About 60% of the 15- to 42-nt reads originated from the coding sequences, in a pattern reminiscent of ribosome profiling. Blood sample type had a considerable influence on the exRNA profile. On average approximately 350-1100 distinct ex-mRNA transcripts were detected depending on plasma type. In serum, additional transcripts from neutrophils and hematopoietic cells increased this number to near 2300. EDTA and ACD plasma showed a destabilizing effect on ex-mRNA and noncoding RNA ribonucleoprotein complexes compared with other plasma types. In a proof-of-concept study, we investigated differences between the exRNA profiles of patients with acute coronary syndrome and healthy controls. The improved tissue resolution of ex-mRNAs after PNK treatment enabled us to detect a neutrophil signature in ACS that escaped detection by ex-miRNA analysis.
Authors with CRIS profile
Kai Hildner
Juniorprofessur für Pneumologie/ Immunologie
Markus Neurath
Lehrstuhl für Innere Medizin I (Medizin 1)
Involved external institutions
Rockefeller University
United States (USA) (US)
Columbia University
United States (USA) (US)
Ruprecht-Karls-Universität Heidelberg
Germany (DE)
United States (USA) (US)
Columbia University
United States (USA) (US)
Ruprecht-Karls-Universität Heidelberg
Germany (DE)
How to cite
APA:
Akat, K.M., Lee, Y.A., Hurley, A., Morozov, P., Max, K.E.A., Brown, M.,... Tuschl, T. (2019). Detection of circulating extracelluar mRNAs by modified small-RNA-sequencing analysis. JCI Insight, 4(9). https://doi.org/10.1172/jci.insight.127317
MLA:
Akat, Kemal M., et al. "Detection of circulating extracelluar mRNAs by modified small-RNA-sequencing analysis." JCI Insight 4.9 (2019).
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