title: An optimized procedure for exosome isolation and analysis using serum samples: Application to cancer biomarker discovery.
abstract: Exosomes are RNA and protein-containing nanovesicles secreted by all cell types and found in abundance in body fluids, including blood, urine and cerebrospinal fluid. These vesicles seem to be a perfect source of biomarkers, as their cargo largely reflects the content of parental cells, and exosomes originating from all organs can be obtained from circulation through minimally invasive or non-invasive means. Here we describe an optimized procedure for exosome isolation and analysis using clinical samples, starting from quick and robust extraction of exosomes with Total exosome isolation reagent, then isolation of RNA followed by qRT-PCR. Effectiveness of this workflow is exemplified by analysis of the miRNA content of exosomes derived from serum samples - obtained from the patients with metastatic prostate cancer, treated prostate cancer patients who have undergone prostatectomy, and control patients without prostate cancer. Three promising exosomal microRNA biomarkers were identified, discriminating these groups: hsa-miR375, hsa-miR21, hsa-miR574.
authors: Thermo Fisher Scientific, Austin, TX 78744, USA.
authors: Mu
authors: M
authors: Li
authors: Department of Pathology and Cell Biology and Center for Advanced Laboratory Medicine, Columbia University Medical Center and New York Presbyterian Hospital, New York, NY, USA.
authors: Alex J
authors: AJ
authors: Rai
authors: Department of Urology, Columbia University Medical Center & New York Presbyterian New York, NY, USA.
authors: G Joel
authors: GJ
authors: DeCastro
authors: Thermo Fisher Scientific, Austin, TX 78744, USA.
authors: Emily
authors: E
authors: Zeringer
authors: Thermo Fisher Scientific, Austin, TX 78744, USA.
authors: Timothy
authors: T
authors: Barta
authors: Thermo Fisher Scientific, Austin, TX 78744, USA.
authors: Susan
authors: S
authors: Magdaleno
authors: Thermo Fisher Scientific, Austin, TX 78744, USA.
authors: Robert
authors: R
authors: Setterquist
authors: Thermo Fisher Scientific, Austin, TX 78744, USA. Electronic address: sasha.vlassov@thermofisher.com.
authors: Alexander V
authors: AV
authors: Vlassov
mesh_terms: D014408:Biomarkers, Tumor / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D016022:Case-Control Studies; D055354:Exosomes / Q000737:chemistry*; D015870:Gene Expression; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D008297:Male; D035683:MicroRNAs / Q000097:blood* / Q000235:genetics; D017430:Prostate-Specific Antigen / Q000097:blood / Q000235:genetics; D011468:Prostatectomy; D011471:Prostatic Neoplasms / Q000737:chemistry* / Q000175:diagnosis / Q000235:genetics / Q000601:surgery; D012334:RNA, Neoplasm / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D014408:Biomarkers, Tumor; D007202:Indicators and Reagents; C524940:MIRN21 microRNA, human; C540933:MIRN375 microRNA, human; C558811:MIRN574 microRNA, human; D035683:MicroRNAs; D012334:RNA, Neoplasm; D017430:Prostate-Specific Antigen
keywords: Biomarker; Blood; Exosome; Extracellular vesicle; Prostate cancer; miRNA; microRNA