Estradiol-Responsive miR-365a-3p Interacts with Tissue Factor 3′UTR to Modulate Tissue Factor-Initiated Thrombin Generation

Jiayin Tian; Murray J. Adams; Jasmine Wee Ting Tay; Ian James; Suzanne Powell; Quintin W. Hughes; Grace Gilmore; Ross I. Baker; Jim Yu-Hsiang Tiao

doi:10.1055/a-1382-9983

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2021; 121(11): 1483-1496
DOI: 10.1055/a-1382-9983

Stroke, Systemic or Venous Thromboembolism

Estradiol-Responsive miR-365a-3p Interacts with Tissue Factor 3′UTR to Modulate Tissue Factor-Initiated Thrombin Generation

Jiayin Tian

¹Western Australian Centre for Thrombosis and Haemostasis, Murdoch University, Murdoch, Perth, Australia

²Perth Blood Institute, West Perth, Perth, Australia

³College of Science, Health, Engineering and Education, Murdoch University, Murdoch, Perth, Australia

⁴Centre for Molecular Medicine and Innovative Therapeutics, Health Futures Institute, Murdoch University, Murdoch, Perth, Australia

,

Murray J. Adams

³College of Science, Health, Engineering and Education, Murdoch University, Murdoch, Perth, Australia

,

Jasmine Wee Ting Tay^*

¹Western Australian Centre for Thrombosis and Haemostasis, Murdoch University, Murdoch, Perth, Australia

²Perth Blood Institute, West Perth, Perth, Australia

,

Ian James

⁵Institute for Immunology and Infectious Diseases, Murdoch University, Murdoch, Perth, Australia

,

Suzanne Powell

¹Western Australian Centre for Thrombosis and Haemostasis, Murdoch University, Murdoch, Perth, Australia

,

Quintin W. Hughes^*

¹Western Australian Centre for Thrombosis and Haemostasis, Murdoch University, Murdoch, Perth, Australia

²Perth Blood Institute, West Perth, Perth, Australia

,

Grace Gilmore

¹Western Australian Centre for Thrombosis and Haemostasis, Murdoch University, Murdoch, Perth, Australia

²Perth Blood Institute, West Perth, Perth, Australia

⁴Centre for Molecular Medicine and Innovative Therapeutics, Health Futures Institute, Murdoch University, Murdoch, Perth, Australia

,

Ross I. Baker

¹Western Australian Centre for Thrombosis and Haemostasis, Murdoch University, Murdoch, Perth, Australia

²Perth Blood Institute, West Perth, Perth, Australia

⁴Centre for Molecular Medicine and Innovative Therapeutics, Health Futures Institute, Murdoch University, Murdoch, Perth, Australia

,

Jim Yu-Hsiang Tiao

¹Western Australian Centre for Thrombosis and Haemostasis, Murdoch University, Murdoch, Perth, Australia

²Perth Blood Institute, West Perth, Perth, Australia

⁴Centre for Molecular Medicine and Innovative Therapeutics, Health Futures Institute, Murdoch University, Murdoch, Perth, Australia

› Author Affiliations

Abstract

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.

Keywords

estrogen - microRNA - thrombin generation - thrombosis - tissue factor

Full Text

References

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Supplementary Material

Supplementary Material