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Potential pathophysiological role of microRNA 193b-5p in human placentae from pregnancies complicated by preeclampsia and intrauterine growth restriction

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Abstract

Preeclampsia (PE) and intrauterine growth restriction (IUGR) are pregnancy complications resulting from abnormal placental development. MicroRNAs can regulate placental development and contribute to disease, by influencing gene expression. Our previous study revealed an increase in miR-193b-5p expression in placentae from patients with early-onset pregnancy complications and identified candidate gene targets for miR-193b-5p. The purpose of this study is two-fold, first to validate candidate gene targets predicted for miR-193b-5p from microRNA-RNA expression data. Second, to overexpress miR-193b-5p in a trophoblast cell line (HTR-8/SVneo) to assess impact on trophoblast cell proliferation and migration. Integration of the miRNA and RNA sequencing expression data revealed 10 candidate gene targets for miR-193b-5p across all patient groups (PE only, IUGR only, PE + IUGR). Luciferase experiments identified two gene targets for miR-193b-5p, APLN and FGF13. Real-time PCR confirmed a median 45% decrease of FGF13 expression across 3 patient groups, and 50% decrease of APLN expression in patients with PE + IUGR. Following transfection of HTR-8/SVneo cells with miR-193b-5p mimics, APLN and FGF13 mRNA expression in HTR-8/SVneo was reduced by a median percentage of 30% and 45%, respectively. Concomitantly, HTR-8/SVneo cells demonstrate 40% reduction in cell migration. APLN and FGF13 immunoreactivity was identified strongly in the cytotrophoblast cells of the human placentae. These findings suggest that miR-193b-5p may contribute to trophoblast dysfunction observed in pregnancy complications such as PE and IUGR.

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Abbreviations

APLN:

Apelin

CT:

Cytotrophoblast

EO:

Early-onset

EVT:

Extravillous trophoblast

FGF13:

Fibroblast growth factor 13

GO:

Gene ontology

IHC:

Immunohistochemistry

IUGR:

Intrauterine growth restriction

miRNA:

MicroRNA

NGS:

Next generation sequencing

PE:

Preeclampsia

qRT-PCR:

Quantitative real time PCR

SCT:

Syncytiotrophoblast

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Acknowledgements

We would like to thank all the donors and the Research Centre for Women’s and Infants Health (RCWIH) BioBank for placental samples used in this project. We would also like to acknowledge Karen Nygard (Biotron Facility, Western University) for assistance with immunohistochemical staining of placental tissues.

Funding

This study was funded by grants from the Canadian Institutes of Health Research (15579 and 15262 to VKMH) and The Douglas and Vivian Bocking Chair in Fetal and Newborn Growth (to VKMH). ZA is supported through Western University’s Graduate Research Scholarship and the Graduate Student Grant from Western University’s Department of Paediatrics.

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Authors and Affiliations

  1. Children’s Health Research Institute, 800 Commissioners Road East, London, ON, N6C 2V5, Canada

    Zain Awamleh & Victor K. M. Han

  2. Department of Biochemistry, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, N6A 3K7, Canada

    Zain Awamleh & Victor K. M. Han

  3. Department of Pediatrics, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, N6A 3K7, Canada

    Victor K. M. Han

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  1. Zain Awamleh
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ZA made substantial contributions to design, acquisition of data, analysis and interpretation of data, and in writing and revising the article. VKMH made substantial contributions to design, interpretation of data and revising the article. All authors approved final version of the article.

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Correspondence to Zain Awamleh.

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Supplementary Figure 1

Negative control images for immunohistochemical analysis. Slides designated negative controls underwent the same procedures, with the exception of the application of the primary antibody. (A) Chorionic villus section and (B) Basal plate decidua section from preterm control, gestational age 29+4. Blue staining is CAT Hematoxylin counterstain. All images were captured at 20 x, bar = 50 m. (TIFF 43115 kb)

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Awamleh, Z., Han, V.K.M. Potential pathophysiological role of microRNA 193b-5p in human placentae from pregnancies complicated by preeclampsia and intrauterine growth restriction. Mol Biol Rep 47, 6531–6544 (2020). https://doi.org/10.1007/s11033-020-05705-y

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