Exosomal miRNA in pre-eclampsia: good things come in small packages

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      Background

      Studies have shown circulating miRNA dysregulation in pre-eclampsia, however difficulties in reproducibility have limited their use as biomarkers. Exosomal miRNA are a promising avenue to identify reproducibly dysregulated miRNA biomarkers for pre-eclampsia.

      Aims

      To optimise exosome isolation from maternal serum in pregnancy and examine serum-derived exosomal miR-1 and miR-210 dysregulation in pre-eclampsia.

      Methods

      Exosome yields from maternal serum isolated by ultracentrifugation and ExoQuickTM isolation methods were compared using electron microscopy and BCA assay-measured protein content. Exosomal RNA was extracted using a modified and manufacturer's mirVanaTM PARISTM protocol and TRIzol® and SeraMirTM protocols. qRT-PCR was performed on SeraMirTM-extracted RNA for U6 small nuclear RNA, miR-1 and miR-210.

      Results

      ExoQuickTM isolated the highest exosome yield as indicated by electron microscopy visualisation and protein content; 11215 μg vs ultracentrifugation ≤12 μg. SeraMirTM extracted the highest and only exosomal RNA concentration above the limit of detection (15.7 ng/μL). No amplification products were detected by qRT-PCR.

      Discussion

      This is the first study to isolate serum-derived exosomes and exosomal RNA in pregnancy and provides a workflow to discover exosomal miRNA biomarkers for pre-eclampsia and other pregnancy complications. Findings suggest more sensitive PCR techniques; pre-amplification or digital PCR, are required to demonstrate exosomal miRNA dysregulation and discover these biomarkers.
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