Reduced Platelet miR-223 Induction in Kawasaki Disease Leads to Severe Coronary Artery Pathology Through a miR-223/PDGFRβ Vascular Smooth Muscle Cell Axis

Abstract

Rationale: Kawasaki disease (KD) is an acute vasculitis of early childhood that can result in per-manent coronary artery structural damage. The etiology for this arterial vulnerability in up to 15% of KD patients is unknown. Vascular smooth muscle cell (VSMC) dedifferentiation play a key role in the pathophysiology of medial damage and aneurysm formation, recognized arterial pathology in KD. Platelet hyperreactivity is also a hallmark of KD. We recently demonstrated that uptake of platelets and platelet-derived miRNAs influences VSMC phenotype in vivo. Objective: We set out to explore whether platelet/vascular smooth muscle cell interactions contrib-ute to coronary pathology in KD. Methods and Results: We prospectively recruited and studied 242 KD patients, 75 of whom had doc-umented coronary artery pathology. Genome-wide miRNA sequencing and droplet digital PCR (ddPCR) demonstrated that KD patient platelets have significant induction of miR-223 compared to healthy controls. Platelet-derived miR-223 has recently been shown to promote vascular smooth muscle quiescence and resolution of wound healing after vessel injury. Paradoxically, KD patients with the most severe coronary pathology (giant coronary artery aneurysms) exhibited a lack of miR-223 induction. Hyperactive platelets isolated from KD patients are readily taken up by VSMCs, delivering functional miR-223 into the VSMCs promoting VSMC differentiation via downregulation of platelet-derived growth factor receptor β (PDGFRβ). The lack of miR-223 induction in patients with severe coronary pathology leads to persistent VSMC dedifferentiation. In a mouse model of KD (Lactobacillus casei cell wall extract (LCWE) injection), miR-223 knockout (miR-223 KO) mice exhibited increased medial thickening, loss of contractile VSMCs in the media, and fragmenta-tion of medial elastic fibers compared to WT mice, which demonstrated significant miR-223 induc-tion upon LCWE challenge. The excessive arterial damage in the miR-223 knockout could be res-cued by adoptive transfer of platelet, administration of miR-223 mimics, or the PDGFRβ inhibitor imatinib mesylate. Interestingly, miR-223 levels progressively increase with age, with the lowest levels found in less than five-year-old. This provides a basis for coronary pathology susceptibility in this very young cohort. Conclusions: Platelet-derived miR-223 (through PDGFRβ inhibition) promotes VSMC differentia-tion and resolution of KD induced vascular injury. Lack of miR-223 induction leads to severe coro-nary pathology characterized by VSMC dedifferentiation and medial damage. Detection of platelet-derived miR-223 in KD patients (at the time of diagnosis) may identify patients at greatest risk of coronary artery pathology. Moreover, targeting platelet miR-223 or VSMC PDGFRβ represents po-tential therapeutic strategies to alleviate coronary pathology in KD.

In silico Identification of 10 Hub Genes and an miRNA–mRNA Regulatory Network in Acute Kawasaki Disease

“…The latter has been verified by RT-PCR to be upregulated in our previous study ( Chen Y. et al, 2018 ). Together with other studies ( Chu et al, 2017 ; Parra-Izquierdo et al, 2020 ; Zhang et al, 2020 ), our recent find ( Wang et al, 2019 ) supported that miR-223 is an important regulator of vascular endothelial cell injury in KD. Thus miR-223 may be a novel target for the treatment of KD.…”

Thrombocytopathy and endotheliopathy: crucial contributors to COVID-19 thromboinflammation

“…Neutrophils and other immune cells enter the vessel wall, promoting endothelial damage and subsequent myofibroblast hyperproliferation 210 . Thrombocytopathy and endotheliopathy are shared factors in the pathogenesis of both COVID-19 and Kawasaki disease 211 . Endothelial function can remain impaired for years after Kawasaki disease resolves 212 .…”

“…Low-dose aspirin has long been used for the treatment of Kawasaki disease, features of which have been associated with COVID-19. In addition, platelets are increasingly recognized to have a role in Kawasaki disease 211 . However, aspirin and other antiplatelet agents do not address endotheliopathy.…”

Role of smooth muscle cells in Cardiovascular Disease

“…For example, Zhang et al reported that without miR-223 VSMC dedifferentiation proceeds undamped resulting in medial damage, apoptosis, and MMP9 release. The elevated MMP9 levels drive the degradation of arterial wall matrix, and subsequently promoting CAA formation [80][81][82]. It is a well-known fact that SMCs senescence is also proven to enhance the formation and rupture of aneurysm as well [83,84].…”