title: Anti-miR-148a regulates platelet FcγRIIA signaling and decreases thrombosis in vivo in mice.
abstract: Fc receptor for IgG IIA (FcγRIIA)-mediated platelet activation is essential in heparin-induced thrombocytopenia (HIT) and other immune-mediated thrombocytopenia and thrombosis disorders. There is considerable interindividual variation in platelet FcγRIIA activation, the reasons for which remain unclear. We hypothesized that genetic variations between FcγRIIA hyper- and hyporesponders regulate FcγRIIA-mediated platelet reactivity and influence HIT susceptibility. Using unbiased genome-wide expression profiling, we observed that human hyporesponders to FcγRIIA activation showed higher platelet T-cell ubiquitin ligand-2 (TULA-2) mRNA expression than hyperresponders. Silent interfering RNA-mediated knockdown of TULA-2 resulted in hyperphosphorylation of spleen tyrosine kinase following FcγRIIA activation in HEL cells. Significantly, we found miR-148a-3p targeted and inhibited both human and mouse TULA-2 mRNA. Inhibition of miR-148a in FcγRIIA transgenic mice upregulated the TULA-2 level and reduced FcγRIIA- and glycoprotein VI-mediated platelet αIIbβ3 activation and calcium mobilization. Anti-miR-148a also reduced thrombus formation following intravascular platelet activation via FcγRIIA. These results show that TULA-2 is a target of miR-148a-3p, and TULA-2 serves as a negative regulator of FcγRIIA-mediated platelet activation. This is also the first study to show the effects of in vivo miRNA inhibition on platelet reactivity. Our work suggests that modulating miR-148a expression is a potential therapeutic approach for thrombosis.
authors.affiliation: Cardeza Foundation for Hematologic Research, Thomas Jefferson University, Philadelphia, PA;
authors.forename: Yuhang
authors.identifier: http://orcid.org/0000-0001-7904-4978
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Cardeza Foundation for Hematologic Research, Thomas Jefferson University, Philadelphia, PA;
authors.forename: Shaji
authors.identifier: http://orcid.org/0000-0002-4182-1855
authors.initials: S
authors.lastname: Abraham
authors.affiliation: Cardeza Foundation for Hematologic Research, Thomas Jefferson University, Philadelphia, PA;
authors.forename: Pierrette
authors.initials: P
authors.lastname: Andre
authors.affiliation: Cardeza Foundation for Hematologic Research, Thomas Jefferson University, Philadelphia, PA;
authors.forename: Leonard C
authors.identifier: http://orcid.org/0000-0002-8976-6410
authors.initials: LC
authors.lastname: Edelstein
authors.affiliation: Baylor College of Medicine, Houston, TX; and.
authors.forename: Chad A
authors.identifier: http://orcid.org/0000-0001-7267-5398
authors.initials: CA
authors.lastname: Shaw
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple University, Philadelphia, PA.
authors.forename: Carol A
authors.initials: CA
authors.lastname: Dangelmaier
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple University, Philadelphia, PA.
authors.forename: Alexander Y
authors.initials: AY
authors.lastname: Tsygankov
authors.affiliation: Sol Sherry Thrombosis Research Center, Temple University, Philadelphia, PA.
authors.forename: Satya P
authors.initials: SP
authors.lastname: Kunapuli
authors.affiliation: Cardeza Foundation for Hematologic Research, Thomas Jefferson University, Philadelphia, PA;
authors.forename: Paul F
authors.initials: PF
authors.lastname: Bray
authors.affiliation: Cardeza Foundation for Hematologic Research, Thomas Jefferson University, Philadelphia, PA;
authors.forename: Steven E
authors.initials: SE
authors.lastname: McKenzie
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000378:metabolism; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D020022:Genetic Predisposition to Disease; D055106:Genome-Wide Association Study; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D035683:MicroRNAs / Q000235:genetics*; D015539:Platelet Activation / Q000235:genetics*; D017027:Protein Tyrosine Phosphatases / Q000096:biosynthesis* / Q000235:genetics; D017452:Receptors, IgG / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013921:Thrombocytopenia / Q000235:genetics; D013927:Thrombosis / Q000235:genetics*
chemical_list: C080568:Fc gamma receptor IIA; C550570:MIRN148 microRNA, human; D035683:MicroRNAs; C554644:Mirn148 microRNA, mouse; D017452:Receptors, IgG; D017027:Protein Tyrosine Phosphatases; C553497:TULA-2 protein, mouse; C489397:UBASH3B protein, human