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Estradiol-responsive miR-365a-3p interacts with tissue factor 3′UTR to modulate tissue factor-initiated thrombin generation

Tian, J.ORCID: 0000-0001-9887-7211, Adams, M.J.ORCID: 0000-0002-7743-4515, Tay, J.W.T., James, I., Powell, S., Hughes, Q.W., Gilmore, G., Baker, R.I. and Tiao, J.Y-H. (2021) Estradiol-responsive miR-365a-3p interacts with tissue factor 3′UTR to modulate tissue factor-initiated thrombin generation. Thrombosis and Haemostasis, 121 (11). pp. 1483-1496.

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Background: High estradiol (E2) levels are linked to an increased risk of venous thromboembolism; however, the underlying molecular mechanism(s) remain poorly understood. We previously identified an E2-responsive microRNA (miR), miR-494–3p, that downregulates protein S expression, and posited additional coagulation factors, such as tissue factor, may be regulated in a similar manner via miRs.

Objectives: To evaluate the coagulation capacity of cohorts with high physiological E2, and to further characterize novel E2-responsive miR and miR regulation on tissue factor in E2-related hypercoagulability.

Methods: Ceveron Alpha thrombin generation assay (TGA) was used to assess plasma coagulation profile of three cohorts. The effect of physiological levels of E2, 10 nM, on miR expression in HuH-7 cells was compared using NanoString nCounter and validated with independent assays. The effect of tissue factor-interacting miR was confirmed by dual-luciferase reporter assays, immunoblotting, flow cytometry, biochemistry assays, and TGA.

Results: Plasma samples from pregnant women and women on the contraceptive pill were confirmed to be hypercoagulable (compared with sex-matched controls). At equivalent and high physiological levels of E2, miR-365a-3p displayed concordant E2 downregulation in two independent miR quantification platforms, and tissue factor protein was upregulated by E2 treatment. Direct interaction between miR-365a-3p and F3-3′UTR was confirmed and overexpression of miR-365a-3p led to a decrease of (1) tissue factor mRNA transcripts, (2) protein levels, (3) activity, and (4) tissue factor-initiated thrombin generation.

Conclusion: miR-365a-3p is a novel tissue factor regulator. High E2 concentrations induce a hypercoagulable state via a miR network specific for coagulation factors.

Item Type: Journal Article
Murdoch Affiliation(s): Centre for Molecular Medicine and Innovative Therapeutics (CMMIT)
Health Futures Institute
Institute for Immunology and Infectious Diseases
Western Australian Centre for Thrombosis and Haemostasis (WACTH)
Publisher: Schattauer
Copyright: © 2021 Thieme
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