Multifunctional Nanoparticles Facilitate Molecular Targeting and miRNA Delivery to Inhibit Atherosclerosis in ApoE^–/– Mice

Abstract

The current study presents an effective and selective multifunctional nanoparticle used to deliver antiatherogenic therapeutics to inflamed pro-atherogenic regions without off-target changes in gene expression or particle-induced toxicities. MicroRNAs (miRNAs) regulate gene expression, playing a critical role in biology and disease including atherosclerosis. While anti-miRNA are emerging as therapeutics, numerous challenges remain due to their potential off-target effects, and therefore the development of carriers for selective delivery to diseased sites is important. Yet, co-optimization of multifunctional nanoparticles with high loading efficiency, a hidden cationic domain to facilitate lysosomal escape and a dense, stable incorporation of targeting moieties is challenging. Here, we create coated, cationic lipoparticles (CCLs), containing anti-miR-712 (∼1400 molecules, >95% loading efficiency) within the core and with a neutral coating, decorated with 5 mol % of peptide (VHPK) to target vascular cell adhesion molecule 1 (VCAM1). Optical imaging validated disease-specific accumulation as anti-miR-712 was efficiently delivered to inflamed mouse aortic endothelial cells in vitro and in vivo. As with the naked anti-miR-712, the delivery of VHPK-CCL-anti-miR-712 effectively downregulated the d-flow induced expression of miR-712 and also rescued the expression of its target genes tissue inhibitor of metalloproteinase 3 (TIMP3) and reversion-inducing-cysteine-rich protein with kazal motifs (RECK) in the endothelium, resulting in inhibition of metalloproteinase activity. Moreover, an 80% lower dose of VHPK-CCL-anti-miR-712 (1 mg/kg dose given twice a week), as compared with naked anti-miR-712, prevented atheroma formation in a mouse model of atherosclerosis. While delivery of naked anti-miR-712 alters expression in multiple organs, miR-712 expression in nontargeted organs was unchanged following VHPK-CCL-anti-miR-712 delivery.

Interleukin-32α Inhibits Endothelial Inflammation, Vascular Smooth Muscle Cell Activation, and Atherosclerosis by Upregulating Timp3 and Reck through suppressing microRNA-205 Biogenesis

“…Importantly, quantitative polymerase chain reaction (qPCR) and immunohistochemistry analysis showed that Timp3 and Reck mRNA and protein expression were significantly decreased in partially-ligated LCA compared with contralateral RCA in non-Tg control mice (Figs. 6A, B), consistent with previous findings 34, 35, but upregulated in the LCA and aorta tissue from hIL-32α-Tg-mice LCA (Figs. 6A, B; Supplementary Fig.…”

HDL and microRNA therapeutics in cardiovascular disease

“…As such, mimics are designed as double-stranded duplexes, consisting of the guide strand identical to the miRNA and a complementary passenger strand that is often modified to increase or shift utilization to the guide strand (van Rooij, Purcell, & Levin, 2012); however, this renders the mimics difficult to transport through cell membranes and they can be degraded in circulation. To circumvent this barrier, several delivery vehicles have been put forward, including DNA vectors and adeno-associated virus (Kota, et al, 2009), lentivirus (Kortylewski & Nechaev, 2014), nanoparticles (Diaz & Vivas-Mejia, 2013; Kheirolomoom, et al, 2015), liposomes (Akinc, et al, 2010; Akinc, et al, 2008; Morrissey, et al, 2005), and stents (D. Wang, et al, 2015).…”

Targeting and therapeutic peptides in nanomedicine for atherosclerosis

“…Kheirolomoom and colleagues used the shortened version of the peptide, VHPK, to coat cationic lipoparticles (CCL) containing anti-miR-712 within its core. 33 miR-712 is a pro-atherogenic, mechanosensitive miRNA that is upregulated by distributed flow in endothelial cells. VHPK-CCLs containing miR-712 inhibitors downregulated the miR and rescued the expression of its target genes, preventing atheroma formation in Apo E −/− mice.…”