Nanoparticle-mediated microRNA-145 Delivery for Vascular Smooth Muscle Cell Phenotype Modulation and Atherosclerosis Treatment

Vascular smooth muscle cells (VSMCs) change from contractile to the synthetic phenotype during atherogenesis and 30-70% of cells that make up plaques have been elucidated to be of VSMC origin. MicroRNA-145 (miR-145) is responsible for regulating VSMC phenotypic switching, and low miR-145 levels in circulation have been linked with atherosclerosis. Hence, we developed nanoparticles for targeted delivery of miR-145 by synthesizing micelles co-assembled with miR-145 and the CCR2-binding peptides for plaque targeting. The miR cargo was protected in micelles from premature endosomal degradation and rescued contractile markers in synthetic VSMCs and SMCs isolated from patient arteries in vitro . In ApoE-/- mid-stage atherosclerotic mice, miR-145 micelles halted plaque growth and maintained contractile phenotypes similar to baseline levels. In early-stage atherosclerosis, a single dose of miR-145 micelles prevented lesion growth by 49%. We present the potential of miR-145 micelles as a therapeutic that can be applied longitudinally and intervene throughout atherosclerosis pathogenesis. ### Competing Interest Statement The authors have declared no competing interest. * VSMCs : vascular smooth muscle cells miR-145 : micelles miR-145 containing peptide amphiphile micelles CCR2 : C-C chemokine receptor 2 MCP-1 : monocyte chemoattractant protein-1 α-SMA : alpha smooth muscle actin miR : microRNA KLF-4/5 : Krüppel-like factor 4 and 5 ELK-1 : ETS domain-containing protein-1 PEG : polyethylene glycol SRF : serum response factor TEM : transmission electron microscopy DLS : dynamic light scattering CMC : critical micelle concentration FBS : fetal bovine serum hASMCs : human aortic smooth muscle cells Me-β-CD : methyl-β-cyclodextrin Cy5miR-145 : micelles Cy5 labeled-miR-145 micelles CT-B : cholera toxin subunit B GSH : glutathione AF647 : Alexa Fluor 647 BUN : blood urea nitrogen SMGS : smooth muscle growth supplement IACUC : Institutional Animal Care and Use Committee HPLC : high performance liquid chromatography HBSS : Hank’s balanced salt solution TBS : tris buffered saline BSA : bovine serum albumin S.D. : standard deviation PEI : polyethylenimine DOTAP : 1,2-dioleoyl-3-trimethylammonium-propane PCL : polycaprolactone DOPE : 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine PLL : poly(L-lysine) DMTAP : dimyristoyltrimethylammonium propane DMPC : dimyristoylphosphatidylcholine BHEM/chol : N,N-bis(2-hydroxyethyl)-N-methyl-N-(2-cholesteryloxycarbonyl aminoethyl) ammonium bromide.