Azadeh Kheirolomoom1*, Chan Woo Kim2*, Jai W. Seo1, Sandeep Kumar2, Dong Ju Son2, M. Karen J. Gagnon1, Elizabeth S. Ingham1, Katherine W. Ferrara1¶, and Hanjoong Jo2,3¶
1Department of Biomedical Engineering, University of California, Davis, CA, USA
2Wallace H. Coulter Department of Biomedical Engineering and 3Division of Cardiology, Georgia
Institute of Technology and Emory University, Atlanta, GA, USA
*AK and CWK contributed equally to this work.
¶KWF and HJ contributed equally to this paper and are co-corresponding authors.
Abstract
The current study presents an effective and selective multi-functional nanoparticle used to deliver anti-atherogenic 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 lipo-particles (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, as compared with naked anti-miR-712 (1 mg/kg dose given twice a week), 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 non-targeted organs was unchanged following VHPK-CCL-anti-miR-712 delivery.
KEY WORDS: microRNA, anti-miRNA, multi-functional particles, targeted delivery, atherosclerosis, endothelial inflammation