Oral Presentation Australia and New Zealand Society for Extracellular Vesicles Conference 2023

Plasma small extracellular vesicle (sEV) derived micro-RNA (miRNA) as biomarkers of fertility; validating miR181b-5p as a plasma sEV biomarker of infertility. (#29)

Pevindu Abeysinghe 1 , Natalie Turner 1 , Holly Flay 2 , Susanne Meier 2 , Leila Cuttle 1 , Murray MItchell 1
  1. Centre for Children’s Health Research, School of Biomedical Sciences. Faculty of Health, The Queensland University of Technology,, Brisbane, QLD, Australia
  2. DairyNZ Limited, Private Bag 3221, Hamilton 3240, New Zealand

Background: The identification of plasma sEV miRNA as robust biomarkers of fertility is limited; however, they are considered as excellent diagnostic and prognostic biomarker candidates. Mir-181b-5p is of particular interest as it is upregulated in inflammatory tumour tissues (1) and demonstrates elevated expression in bovine endometrial stromal cells prior to decidualization (2).

Methods: Plasma samples were collected from 10-month-old Holstein-Friesian (Bos taurus) dairy cows of distinct fertility; High fertile (HF) and low fertile (LF) groups (n=8 each). ExoEasy Maxi and Midi spin columns (77023, QIAGEN) were used to isolate plasma sEV total RNA. Real-time PCR was conducted using miRCURY LNA miRNA PCR Assays (339306, QIAGEN) for the previously identified differentially expressed (DE) miRNAs between HF and LF groups.

Results: The expression of miR-181b-5p was upregulated in LF sEV, in contrast to HF sEV (P value = 0.0093, Fold change = 2.665). Interestingly, bovine endometrial cells demonstrated extremely high levels of miR-181b-5p, similar to Graham et al (2), in contrast to the expression levels in the blood plasma sEV.  MiR-12054 and miR-2285aw were upregulated in LF sEV, and miR-17-5p, miR-2285dd and miR-2335 were upregulated in HF sEV; however, these upregulations were not statistically significant.

Conclusion: MiR-181b-5p was validated as a prognostic sEV miRNA biomarker of infertility, exhibiting upregulation in LF animals as a subsequent pro-inflammatory response.

 

  1. Fahim SA, Abdullah MS, Espinoza-Sánchez NA, Hassan H, Ibrahim AM, Ahmed SH, et al. Inflammatory Breast Carcinoma: Elevated microRNA miR-181b-5p and Reduced miR-200b-3p, miR-200c-3p, and miR-203a-3p Expression as Potential Biomarkers with Diagnostic Value. Biomolecules. 2020;10(7):1059. PubMed PMID: doi:10.3390/biom10071059.
  2. Graham A, Holbert J, Nothnick WB. miR-181b-5p Modulates Cell Migratory Proteins, Tissue Inhibitor of Metalloproteinase 3, and Annexin A2 During In Vitro Decidualization in a Human Endometrial Stromal Cell Line. Reprod Sci. 2017;24(9):1264-74. Epub 2016/01/01. doi: 10.1177/1933719116682877. PubMed PMID: 28256954; PubMed Central PMCID: PMCPMC6344830.