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

Authors

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

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

References
1 Parker MG. Transcriptional activation by oestrogen receptors. Biochem Soc Symp 1998; 63: 45-50
2 Bleker SM, Coppens M, Middeldorp S. Sex, thrombosis and inherited thrombophilia. Blood Rev 2014; 28 (03) 123-133
3 Gialeraki A, Valsami S, Pittaras T, Panayiotakopoulos G, Politou M. Oral contraceptives and HRT risk of thrombosis. Clin Appl Thromb Hemost 2018; 24 (02) 217-225
4 Beyer-Westendorf J, Bauersachs R, Hach-Wunderle V, Zotz RB, Rott H. Sex hormones and venous thromboembolism - from contraception to hormone replacement therapy. Vasa 2018; 47 (06) 441-450
5 Australian Institute of Health Welfare. Maternal deaths in Australia 2016. Canberra: AIHW; 2018. . Accessed February 19, 2021 at: https://www.aihw.gov.au/getmedia/558ae883-a888-406a-b48f-71f562db3918/aihw-per-99-printable-PDF-of-web-report.pdf.aspx
6 Abe K, Kuklina EV, Hooper WC, Callaghan WM. Venous thromboembolism as a cause of severe maternal morbidity and mortality in the United States. Semin Perinatol 2019; 43 (04) 200-204
7 Klinge CM. Estrogen receptor interaction with estrogen response elements. Nucleic Acids Res 2001; 29 (14) 2905-2919
8 Di Bitondo R, Hall AJ, Peake IR, Iacoviello L, Winship PR. Oestrogenic repression of human coagulation factor VII expression mediated through an oestrogen response element sequence motif in the promoter region. Hum Mol Genet 2002; 11 (07) 723-731
9 Adams B, Western AK, Winship PR. Identification and functional characterization of a polymorphic oestrogen response element in the human coagulation factor IX gene promoter. Br J Haematol 2008; 140 (02) 241-249
10 Farsetti A, Narducci M, Moretti F. et al. Inhibition of ERalpha-mediated trans-activation of human coagulation factor XII gene by heteromeric transcription factor NF-Y. Endocrinology 2001; 142 (08) 3380-3388
11 Ali HO, Stavik B, Myklebust CF. et al. Oestrogens downregulate tissue factor pathway inhibitor through oestrogen response elements in the 5′-flanking region. PLoS One 2016; 11 (03) e0152114
12 Castellano L, Giamas G, Jacob J. et al. The estrogen receptor-alpha-induced microRNA signature regulates itself and its transcriptional response. Proc Natl Acad Sci U S A 2009; 106 (37) 15732-15737
13 Jia J, Zhou H, Zeng X, Feng S. Estrogen stimulates osteoprotegerin expression via the suppression of miR-145 expression in MG-63 cells. Mol Med Rep 2017; 15 (04) 1539-1546
14 Bartel DP. Metazoan microRNAs. Cell 2018; 173 (01) 20-51
15 Wightman B, Ha I, Ruvkun G. Posttranscriptional regulation of the heterochronic gene lin-14 by lin-4 mediates temporal pattern formation in C. elegans. Cell 1993; 75 (05) 855-862
16 Ma YS, Lv ZW, Yu F. et al. MicroRNA-302a/d inhibits the self-renewal capability and cell cycle entry of liver cancer stem cells by targeting the E2F7/AKT axis. J Exp Clin Cancer Res 2018; 37 (01) 252
17 Li T, Li M, Xu C. et al. miR–146a regulates the function of Th17 cell differentiation to modulate cervical cancer cell growth and apoptosis through NF–κB signaling by targeting TRAF6. Oncol Rep 2019; 41 (05) 2897-2908
18 Tay J, Tiao J, Hughes Q, Jorritsma J, Gilmore G, Baker R. Circulating microRNA as thrombosis sentinels: caveats and considerations. Semin Thromb Hemost 2018; 44 (03) 206-215
19 Li NX, Sun JW, Yu LM. Evaluation of the circulating MicroRNA-495 and Stat3 as prognostic and predictive biomarkers for lower extremity deep venous thrombosis. J Cell Biochem 2018; 119 (07) 5262-5273
20 Kanuri SH, Ipe J, Kassab K. et al. Next generation microRNA sequencing to identify coronary artery disease patients at risk of recurrent myocardial infarction. Atherosclerosis 2018; 278: 232-239
21 Klinge CM. miRNAs regulated by estrogens, tamoxifen, and endocrine disruptors and their downstream gene targets. Mol Cell Endocrinol 2015; 418 (Pt 3): 273-297
22 Howard EW, Yang X. MicroRNA regulation in estrogen receptor-positive breast cancer and endocrine therapy. Biol Proced Online 2018; 20 (01) 17
23 Tay JW, Romeo G, Hughes QW, Baker RI. Micro-ribonucleic Acid 494 regulation of protein S expression. J Thromb Haemost 2013; 11 (08) 1547-1555
24 Ali HO, Arroyo AB, González-Conejero R. et al. The role of microRNA-27a/b and microRNA-494 in estrogen-mediated downregulation of tissue factor pathway inhibitor α. J Thromb Haemost 2016; 14 (06) 1226-1237
25 Wedderburn RWM. Quasi-likelihood functions, generalized linear models, and the Gauss-Newton method. Biometrika 1974; 61 (03) 439-447
26 Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 2008; 3 (06) 1101-1108
27 Lewis BP, Burge CB, Bartel DP. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 2005; 120 (01) 15-20
28 Betel D, Wilson M, Gabow A, Marks DS, Sander C. The microRNA.org resource: targets and expression. Nucleic Acids Res 2008; 36 (Database issue): D149-D153
29 Huang HY, Chien CH, Jen KH, Huang HD. RegRNA: an integrated web server for identifying regulatory RNA motifs and elements. Nucleic Acids Res 2006; 34 (Web Server issue): W429-34
30 Wong N, Wang X. miRDB: an online resource for microRNA target prediction and functional annotations. Nucleic Acids Res 2015; 43 (Database issue): D146-D152
31 Rosenkranz A, Hiden M, Leschnik B. et al. Calibrated automated thrombin generation in normal uncomplicated pregnancy. Thromb Haemost 2008; 99 (02) 331-337
32 McLean KC, Bernstein IM, Brummel-Ziedins KE. Tissue factor-dependent thrombin generation across pregnancy. Am J Obstet Gynecol 2012; 207 (02) 135.e1-135.e6
33 Kovac MK, Lalic-Cosic SZ, Dmitrovic JM, Djordjevic VJ, Radojkovic DP. Thrombin generation, D-dimer and protein S in uncomplicated pregnancy. Clin Chem Lab Med 2015; 53 (12) 1975-1979
34 Westhoff CL, Pike MC, Cremers S, Eisenberger A, Thomassen S, Rosing J. Endogenous thrombin potential changes during the first cycle of oral contraceptive use. Contraception 2017; 95 (05) 456-463
35 Adams BD, Furneaux H, White BA. The micro-ribonucleic acid (miRNA) miR-206 targets the human estrogen receptor-alpha (ERalpha) and represses ERalpha messenger RNA and protein expression in breast cancer cell lines. Mol Endocrinol 2007; 21 (05) 1132-1147
36 Iorio MV, Ferracin M, Liu CG. et al. MicroRNA gene expression deregulation in human breast cancer. Cancer Res 2005; 65 (16) 7065-7070
37 McFall T, McKnight B, Rosati R. et al. Progesterone receptor A promotes invasiveness and metastasis of luminal breast cancer by suppressing regulation of critical microRNAs by estrogen. J Biol Chem 2018; 293 (04) 1163-1177
38 Kodahl AR, Lyng MB, Binder H. et al. Novel circulating microRNA signature as a potential non-invasive multi-marker test in ER-positive early-stage breast cancer: a case control study. Mol Oncol 2014; 8 (05) 874-883
39 Klinge CM. Estrogen regulation of microRNA expression. Curr Genomics 2009; 10 (03) 169-183
40 Bhat-Nakshatri P, Wang G, Collins NR. et al. Estradiol-regulated microRNAs control estradiol response in breast cancer cells. Nucleic Acids Res 2009; 37 (14) 4850-4861
41 Masri S, Liu Z, Phung S, Wang E, Yuan Y-C, Chen S. The role of microRNA-128a in regulating TGFbeta signaling in letrozole-resistant breast cancer cells. Breast Cancer Res Treat 2010; 124 (01) 89-99
42 Tay J, Tiao J, Hughes Q, Gilmore G, Baker R. Therapeutic potential of miR-494 in thrombosis and other diseases: a review. Aust J Chem 2016; 69 (10) 1078-1093
43 Teruel R, Pérez-Sánchez C, Corral J. et al. Identification of miRNAs as potential modulators of tissue factor expression in patients with systemic lupus erythematosus and antiphospholipid syndrome. J Thromb Haemost 2011; 9 (10) 1985-1992
44 Sahu A, Jha PK, Prabhakar A. et al. MicroRNA-145 impedes thrombus formation via targeting tissue factor in venous thrombosis. EBioMedicine 2017; 26: 175-186
45 Seitz H. Issues in current microRNA target identification methods. RNA Biol 2017; 14 (07) 831-834
46 Osman A, Fälker K. Characterization of human platelet microRNA by quantitative PCR coupled with an annotation network for predicted target genes. Platelets 2011; 22 (06) 433-441
47 Chen YC, Lin FY, Lin YW. et al. Platelet microRNA 365-3p expression correlates with high on-treatment platelet reactivity in coronary artery disease patients. Cardiovasc Drugs Ther 2019; 33 (02) 129-137
48 Guo D, Liu J, Wang W. et al. Alteration in abundance and compartmentalization of inflammation-related miRNAs in plasma after intracerebral hemorrhage. Stroke 2013; 44 (06) 1739-1742
49 Adesanya MA, Maraveyas A, Madden L. Differing mechanisms of thrombin generation in live haematological and solid cancer cells determined by calibrated automated thrombography. Blood Coagul Fibrinolysis 2017; 28 (08) 602-611
50 Marchetti M, Diani E, ten Cate H, Falanga A. Characterization of the thrombin generation potential of leukemic and solid tumor cells by calibrated automated thrombography. Haematologica 2012; 97 (08) 1173-1180

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