A total transcriptome profiling method for plasma-derived extracellular vesicles: applications for liquid biopsies

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Amorim, Maria G. ^[1] ; Valieris, Renan ^[2] ; Drummond, Rodrigo D. ^[2] ; Pizzi, Melissa P. ^[1] ; Freitas, Vanessa M. ^[3] ; Sinigaglia-Coimbra, Rita ^[4] ; Calin, George A. ^{[5, 6]} ; Pasqualini, Renata ^{[7, 8, 9]} ; Arap, Wadih ^{[7, 8, 9]} ; Silva, Israel T. ^{[10, 2]} ; Dias-Neto, Emmanuel ^{[1, 11]} ; Nunes, Diana N. ^[1]

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^[1] AC Camargo Canc Ctr, Lab Med Genom, Sao Paulo, SP - Brazil

^[2] AC Camargo Canc Ctr, Lab Computat Biol, Sao Paulo, SP - Brazil

^[3] Univ Sao Paulo, Inst Biomed Sci, Dept Cell & Dev Biol, Sao Paulo, SP - Brazil

^[4] Univ Fed Sao Paulo, Electron Microscopy Ctr, Sao Paulo, SP - Brazil

^[5] Univ Texas MD Anderson Canc Ctr, Dept Expt Therapeut, Houston, TX 77030 - USA

^[6] Univ Texas MD Anderson Canc Ctr, Ctr RNA Interference & Non Coding RNAs, Houston, TX 77030 - USA

^[7] Univ New Mexico, Comprehens Canc Ctr, Albuquerque, NM 87131 - USA

^[8] Univ New Mexico, Sch Med, Div Hematol Oncol, Dept Internal Med, Albuquerque, NM 87131 - USA

^[9] Univ New Mexico, Sch Med, Div Mol Med, Dept Internal Med, Albuquerque, NM 87131 - USA

^[10] Rockefeller Univ, Lab Mol Immunol, 1230 York Ave, New York, NY 10021 - USA

^[11] FMUSP, Lab Neurociencias Alzira Denise Hertzog Silva LIM, Inst Psiquiatria, Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 7, OCT 31 2017.
• Citações Web of Science: 13

Resumo

Extracellular vesicles (EVs) are key mediators of intercellular communication. Part of their biological effects can be attributed to the transfer of cargos of diverse types of RNAs, which are promising diagnostic and prognostic biomarkers. EVs found in human biofluids are a valuable source for the development of minimally invasive assays. However, the total transcriptional landscape of EVs is still largely unknown. Here we develop a new method for total transcriptome profiling of plasma-derived EVs by next generation sequencing (NGS) from limited quantities of patient-derived clinical samples, which enables the unbiased characterization of the complete RNA cargo, including both small- and long-RNAs, in a single library preparation step. This approach was applied to RNA extracted from EVs isolated by ultracentrifugation from the plasma of five healthy volunteers. Among the most abundant RNAs identified we found small RNAs such as tRNAs, miRNAs and miscellaneous RNAs, which have largely unknown functions. We also identified protein-coding and long noncoding transcripts, as well as circular RNA species that were also experimentally validated. This method enables, for the first time, the full spectrum of transcriptome data to be obtained from minute patient-derived samples, and will therefore potentially allow the identification of cell-to-cell communication mechanisms and biomarkers. (AU)

• Processo FAPESP: 14/26897-0 - Epidemiologia e genômica de adenocarcinomas gástricos no Brasil
• Beneficiário: Emmanuel Dias-Neto
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 11/09172-3 - Avaliação de micropartículas circulantes em pacientes com câncer de mama ductal invasivo Her2+ e sua possível relação com a agressividade tumoral
• Beneficiário: Emmanuel Dias-Neto
• Modalidade de apoio: Auxílio à Pesquisa - Regular