Small-Extracellular-Vesicle-Derived miRNA Profile Identifies miR-483-3p and miR-326 as Regulators in the Pathogenesis of Antiphospholipid Syndrome (APS)

Abstract

Primary antiphospholipid syndrome (PAPS) is a systemic autoimmune disease associated with recurrent thrombosis and/or obstetric morbidity with persistent antiphospholipid antibodies (aPL). Although these antibodies drive endothelial injury and thrombophilia, the underlying molecular mechanism is still unclear. Small extracellular vesicles (sEVs) contain miRNAs, key players in intercellular communication. To date, the effects of miRNA-derived sEVs in PAPS are not well understood. We characterised the quantity, cellular origin and miRNA profile of sEVs isolated from thrombotic APS patients (PAPS, n = 50), aPL-carrier patients (aPL, n = 30) and healthy donors (HD, n = 30). We found higher circulating sEVs mainly of activated platelet origin in PAPS and aPL patients compared to HD, that were highly engulfed by HUVECs and monocyte. Through miRNA-sequencing analysis, we identified miR-483-3p to be differentially upregulated in sEVs from patients with PAPS and aPL, and miR-326 to be downregulated only in PAPS sEVs. In vitro studies showed that miR-483-3p overexpression in endothelial cells induced an upregulation of the PI3K-AKT pathway that led to endothelial proliferation/dysfunction. MiR-326 downregulation induced NOTCH pathway activation in monocytes with the upregulation of NFKB1, tissue factor and cytokine production. These results provide evidence that miRNA-derived sEVs contribute to APS pathogenesis by producing endothelial cell proliferation, monocyte activation and adhesion/procoagulant factors.

Role of microRNAs in Immune Regulation with Translational and Clinical Applications

“…miR-17-92 (rs4284505) [113], miR-146a [114], miR-365a-3p [115], miR-4689 [116] miR-21 [117], miR-124-3p, miR-146, miR-320b, miR-377-3p [118,119] miR-181a, miR-223 [120], miR-485-5p [121] miR-7 [122], miR-30a [123], miR-181d-5p [124], miR-590-3p [125] Antiphospholipid syndrome miR-326, miR-483-3p [126] miR-19b-3p, miR-20a-5p [127] miR-146a-5p [128] Systemic sclerosis, scleroderma miR-21 [129], miR-126 [130], miR-155-5p [131] miR-21 [132], miR-27a [133], miR-29a, miR-138 [134] miR-21-5p, miR-29a-3p, miR-143, miR-155 [135], miR-181b-5p, miR-210-3p, miR-223-3p [136] miR-27a [137], miR-214 [138] Sjögren's syndrome miR-181d-5p [139], miR-223-3p [140] miR-30 [141], miR-181b-5p, miR-322-5p, miR-424-5p, miR-503-5p [142], miR-513c-3p [143], miR-1290, let-7b-5p [144] miR-92a [145] miR-16, miR-125b [146], miR-142, miR-223 [147], let-7 [148] Autoimmune vasculitis miR-424-5p [149] miR-24-3p [150], miR-30a-5p, miR-33, miR-34 [151], miR-99a-5p, miR-106b-5p, miR-133a [152], miR-146a-5p [149], miR-148b-3p, miR-155-5p, miR-182-5p…”

“…Exosomal miR-146a-5p derived from human umbilical cord MSCs can alleviate trophoblast injury and placental dysfunction by regulating the tumor necrosis factor receptor-associated factor 6 (TRAF6)/NFκB axis [128]. Small extracellular vesicles (sEVs) containing miRNAs are involved in the pathogenesis of APS, such as the up-regulation of miR-483-3p and down-regulation of miR-326 through the regulation of phosphatidylinositol-3-kinase (PI3K)-AKT and Notch pathways, respectively [126].…”

MiRNAs and Their Role in Venous Thromboembolic Complications

“…It has been proven that miRNAs play a significant role in various biological processes, including the cell cycle, apoptosis, proliferation, and differentiation, regulating the expression of about 90% of all human genes. In addition, their direct role in the pathogenesis of many human diseases, including the imbalance of the hemostatic system, such as thrombosis, has already been proven (Table 1) [7][8][9][10][11][12][13][14][15][16]. Promotes atherosclerotic plaque rupture and thrombosis [15] Catheter-related thrombosis miR-92a-3p Down In vivo MAPK/NF-κB Inhibits oxidative stress injury and prevents catheter-related thrombosis formation [16] Abbreviations: AAF, Autogenous arteriovenous fistula; COVID-19, Coronavirus disease 2019; VEGF, Vascular endothelial growth factor; Ang-II, Angiotensin II; RHOB, Rho GTPase family member B; P38MAPK, p38 mitogenactivated protein kinases; COX-2, Cyclooxygenase-2; EGFR, epidermal growth factor receptor; HSP90AA1, Heat shock protein 90 alpha family class A member 1; APP, Amyloid-beta precursor protein; PTEN, Phosphatase and tensin homolog; UBC, Ubiquitin C; FN1, Fibronectin 1; ELAVL1, ELAV-like protein 1; CALM1, Calmodulin 1; TF, Tissue factor; EPCR, Endothelial protein C receptor; ERK 1/2, Extracellular signal-regulated kinase; MAPK, p38 mitogen activated protein kinase; NF-κB, Nuclear factor-κB; TNF-α, Tumor necrosis factor alpha; IGF-1R, insulin-like growth factor 1; PI3K, Phosphoinositide 3-kinases; NOTCH, Neurogenic locus notch homolog protein 1; Bcl2, B-cell lymphoma 2.…”