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Protocol for serum exosomal miRNAs analysis in prostate cancer patients treated with radiotherapy

Overview of attention for article published in Journal of Translational Medicine, August 2018
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Title
Protocol for serum exosomal miRNAs analysis in prostate cancer patients treated with radiotherapy
Published in
Journal of Translational Medicine, August 2018
DOI 10.1186/s12967-018-1592-6
Pubmed ID
Authors

Bijaya Malla, Daniel M. Aebersold, Alan Dal Pra

Abstract

Circulating exosomes from prostate cancer (PCa) patients undergoing radiotherapy are attractive candidate biomarkers for monitoring treatment response. Multiple workflows for isolation and content characterization of exosomes in biofluids have been attempted. We report a protocol to isolate and characterize exosomal miRNAs content and assess radiation-induced changes. In this pilot study, we performed targeted exosomal miRNA profiling of 25 serum samples obtained from PCa patients with intermediate- and high-risk disease treated with curative radiotherapy (RT), and controls. Post-treatment blood samples were collected at least 28 days after radiation therapy as a paired follow-up sample. The complete workflow consisted of two phases: I) filtration and polyethylene glycol salt precipitation phase which enriched particles below 200 nm in size followed by characterization using electron microscopy, and II) flow cytometry. Finally, miRNA expression analysis between untreated and treated patient samples was performed using RNA extraction kit, and qRT-PCR. In our preliminary data, 1 ml of serum from PCa patients showed higher exosomal concentration (3.68E+10) compared to controls (6.07E+08). The overall expression of exosomes after RT was found to be higher compared to untreated samples; the median value changed from 3.68E+10 to 5.40E+10; p = 0.52. Using electron microscopy, we were able to visualize cup-shaped vesicles with morphology and size compatible with exosomes. The bead-based flow cytometry showed positivity for exosomal tetraspanins surface markers CD63 and CD9. All five miRNAs (hsa-let-7a-5p, hsa-miR-141-3p, hsa-miR-145-5p, hsa-miR-21-5p, hsa-miR-99b-5p) have been identified in exosomes. Despite overall changes in hsa-let-7a-5p expression after radiation, the difference was significant only in the high-risk group (p = 0.037). In addition, the radiation response to hsa-miR-21-5p was elevated in the high-risk group compared to the intermediate group (p = 0.036). Herewith, we demonstrated a protocol for isolation of serum exosomes and exosomal miRNA amplification. The recovery of exosomal miRNAs and their differential expression after radiation treatment suggests promising biomarker potential that requires further investigation in larger patient cohorts.

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Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 67 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 67 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 14 21%
Student > Ph. D. Student 8 12%
Student > Bachelor 6 9%
Student > Master 6 9%
Student > Doctoral Student 5 7%
Other 8 12%
Unknown 20 30%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 13 19%
Medicine and Dentistry 11 16%
Agricultural and Biological Sciences 6 9%
Computer Science 2 3%
Immunology and Microbiology 2 3%
Other 11 16%
Unknown 22 33%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 14 August 2018.
All research outputs
#20,529,980
of 23,099,576 outputs
Outputs from Journal of Translational Medicine
#3,359
of 4,054 outputs
Outputs of similar age
#288,772
of 330,840 outputs
Outputs of similar age from Journal of Translational Medicine
#61
of 83 outputs
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