Detection of circulating extracellular mRNAs by modified small-RNA-sequencing analysis.

Akat, Kemal M; Lee, Youngmin A; Hurley, Arlene; Morozov, Pavel; Max, Klaas Ea; Brown, Miguel; Bogardus, Kimberly; Sopeyin, Anuoluwapo; Hildner, Kai; Diacovo, Thomas G; Neurath, Markus F; Borggrefe, Martin; Tuschl, Thomas

Akat, Kemal M; Lee, Youngmin A; Hurley, Arlene; Morozov, Pavel; Max, Klaas Ea; Brown, Miguel; Bogardus, Kimberly; Sopeyin, Anuoluwapo; Hildner, Kai; Diacovo, Thomas G; Neurath, Markus F; Borggrefe, Martin; Tuschl, Thomas.

Afiliación

Akat KM; Laboratory of RNA Molecular Biology and.
Lee YA; Laboratory of RNA Molecular Biology and.
Hurley A; Center for Translational Science, The Rockefeller University, New York, New York, USA.
Morozov P; Laboratory of RNA Molecular Biology and.
Max KE; Laboratory of RNA Molecular Biology and.
Brown M; Laboratory of RNA Molecular Biology and.
Bogardus K; Laboratory of RNA Molecular Biology and.
Sopeyin A; Laboratory of RNA Molecular Biology and.
Hildner K; Department of Medicine 1, University Hospital Erlangen, University of Erlangen-Nuremberg, Kussmaul Campus for Medical Research, Erlangen, Bavaria, Germany.
Diacovo TG; Departments of Pediatrics and Cell Biology and Pathology, Columbia University Medical Center, New York, New York, USA.
Neurath MF; Department of Medicine 1, University Hospital Erlangen, University of Erlangen-Nuremberg, Kussmaul Campus for Medical Research, Erlangen, Bavaria, Germany.
Borggrefe M; First Department of Medicine, University Medical Center Mannheim, Faculty of Medicine Mannheim, University of Heidelberg, European Center for AngioScience, and DZHK (German Center for Cardiovascular Research), partner site Heidelberg/Mannheim, Mannheim, Baden-Wuerttemberg, Germany.
Tuschl T; Laboratory of RNA Molecular Biology and.

JCI Insight ; 52019 04 11.

Article en En | MEDLINE | ID: mdl-30973829

RESUMEN

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 exRNA isolated from serum and platelet-poor EDTA, ACD, and heparin plasma to study the effect on ex-mRNA capture. Compared to conventional sRNA-seq PNK-treatment increased the detection of informative ex-mRNAs reads up to 50-fold. The exRNA pool was dominated by 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 to 1,100 distinct ex-mRNA transcripts were detected depending on plasma type. In serum, additional transcripts from neutrophils and hematopoietic cells increased this number to near 2,300. EDTA and ACD plasma showed a destabilizing effect on ex mRNA and non-coding RNA ribonucleoprotein complexes compared to other plasma types. In a proof-of-concept study, we investigated differences between the exRNA profiles of patients with acute coronary syndrome (ACS) 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.

Asunto(s)

Síndrome Coronario Agudo/genética; Células Sanguíneas/metabolismo; Ácidos Nucleicos Libres de Células/genética; Hígado/metabolismo; Músculo Esquelético/metabolismo; RNA-Seq/métodos; Síndrome Coronario Agudo/sangre; Adulto; Anciano; Biomarcadores/sangre; Estudios de Casos y Controles; Ácidos Nucleicos Libres de Células/sangre; Ácido Cítrico; Ácido Edético; Eritrocitos/metabolismo; Femenino; Glucosa/análogos & derivados; Heparina; Secuenciación de Nucleótidos de Alto Rendimiento; Humanos; Leucocitos Mononucleares/metabolismo; Masculino; Persona de Mediana Edad; Monocitos/metabolismo; Neutrófilos/metabolismo; Plasma; Polinucleótido 5'-Hidroxil-Quinasa; Prueba de Estudio Conceptual; Análisis de Secuencia de ARN/métodos; Suero; Manejo de Especímenes

Palabras clave

Bioinformatics; Cardiology; Molecular diagnosis; RNA processing; Vascular Biology

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Sanguíneas / Músculo Esquelético / Síndrome Coronario Agudo / Ácidos Nucleicos Libres de Células / RNA-Seq / Hígado Tipo de estudio: Diagnostic_studies / Observational_studies Límite: Adult / Aged / Female / Humans / Male / Middle aged Idioma: En Revista: JCI Insight Año: 2019 Tipo del documento: Article Pais de publicación: Estados Unidos

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