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Expression of microRNAs in plasma and in extracellular vesicles derived from plasma for dogs with glioma and dogs with other brain diseases

Momoko Narita DVM1, Hidetaka Nishida DVM, PhD1, Ryota Asahina DVM, PhD2, Kohei Nakata DVM2, Hirohito Yano MD, PhD3, Peter J. Dickinson BVSc, PhD5, Toshiyuki Tanaka DVM, PhD6, Hideo Akiyoshi DVM, PhD6, Sadatoshi Maeda DVM, PhD1,2, and Hiroaki Kamishina DVM, PhD1,2,4
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  • 1 1Joint Department of Veterinary Medicine, Faculty of Applied Biological Sciences, Gifu University, 501–1193 Gifu, Japan.
  • | 2 2United Graduate School of Veterinary Sciences, Gifu University, 501–1193 Gifu, Japan.
  • | 3 3Department of Neurosurgery, Graduate School of Medicine, Gifu University, 501–1193 Gifu, Japan.
  • | 4 4Center for Highly Advanced Integration of Nano and Life Sciences, Gifu University, 501–1193 Gifu, Japan.
  • | 5 5Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California-Davis, Davis, CA 95616.
  • | 6 6Department of Advanced Clinical Medicine, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, 598–8531 Osaka, Japan.

Abstract

OBJECTIVE

To measure expression of microRNAs (miRNAs) in plasma and in extracellular vesicles (EVs) derived from plasma for dogs with glioma and dogs with other brain diseases.

SAMPLE

Plasma samples from 11 dogs with glioma and 19 control dogs with various other brain diseases.

PROCEDURES

EVs were isolated from plasma samples by means of ultracentrifugation. Expression of 4 candidate reference miRNAs (let-7a, miR-16, miR-26a, and miR-103) and 4 candidate target miRNAs (miR-15b, miR-21, miR-155, and miR-342-3p) was quantified with reverse transcription PCR assays. Three software programs were used to select the most suitable reference miRNAs from among the 4 candidate reference miRNAs. Expression of the 4 target miRNAs was then calculated relative to expression of the reference genes in plasma and EVs, and relative expression was compared between dogs with glioma and control dogs with other brain diseases.

RESULTS

The most suitable reference miRNAs were miR-16 for plasma and let-7a for EVs. Relative expression of miR-15b in plasma and in EVs was significantly higher in dogs with glioma than in control dogs. Relative expression of miR-342-3p in EVs was significantly higher in dogs with glioma than in control dogs.

CONCLUSIONS AND CLINICAL RELEVANCE

Results suggested that miR-15b and miR-342-3p have potential as noninvasive biomarkers for differentiating glioma from other intracranial diseases in dogs. However, more extensive analysis of expression in specific glioma subtypes and grades, compared with expression in more defined control populations, will be necessary to assess their clinical relevance.

Abstract

OBJECTIVE

To measure expression of microRNAs (miRNAs) in plasma and in extracellular vesicles (EVs) derived from plasma for dogs with glioma and dogs with other brain diseases.

SAMPLE

Plasma samples from 11 dogs with glioma and 19 control dogs with various other brain diseases.

PROCEDURES

EVs were isolated from plasma samples by means of ultracentrifugation. Expression of 4 candidate reference miRNAs (let-7a, miR-16, miR-26a, and miR-103) and 4 candidate target miRNAs (miR-15b, miR-21, miR-155, and miR-342-3p) was quantified with reverse transcription PCR assays. Three software programs were used to select the most suitable reference miRNAs from among the 4 candidate reference miRNAs. Expression of the 4 target miRNAs was then calculated relative to expression of the reference genes in plasma and EVs, and relative expression was compared between dogs with glioma and control dogs with other brain diseases.

RESULTS

The most suitable reference miRNAs were miR-16 for plasma and let-7a for EVs. Relative expression of miR-15b in plasma and in EVs was significantly higher in dogs with glioma than in control dogs. Relative expression of miR-342-3p in EVs was significantly higher in dogs with glioma than in control dogs.

CONCLUSIONS AND CLINICAL RELEVANCE

Results suggested that miR-15b and miR-342-3p have potential as noninvasive biomarkers for differentiating glioma from other intracranial diseases in dogs. However, more extensive analysis of expression in specific glioma subtypes and grades, compared with expression in more defined control populations, will be necessary to assess their clinical relevance.

Supplementary Materials

    • Supplementary Table 1 (PDF 223 kb)
    • Supplementary Table 2 (PDF 221 kb)

Contributor Notes

Dr. Nishida's present address is the Department of Advanced Clinical Medicine, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, 598–8531 Osaka, Japan.

Address correspondence to Dr. Nishida (hnishida@vet.osakafu-u.ac.jp).