Nanocarriers of shRNA-Runx2 directed to collagen IV as a nanotherapeutic system to target calcific aortic valve disease

Geanina Voicu; Cristina Ana Mocanu; Florentina Safciuc; Maria Anghelache; Mariana Deleanu; Sergiu Cecoltan; Mariana Pinteala; Cristina Mariana Uritu; Ionel Droc; Maya Simionescu; Ileana Manduteanu; Manuela Calin

doi:10.1016/j.mtbio.2023.100620

Nanocarriers of shRNA-Runx2 directed to collagen IV as a nanotherapeutic system to target calcific aortic valve disease

Mater Today Bio. 2023 Mar 28:20:100620. doi: 10.1016/j.mtbio.2023.100620. eCollection 2023 Jun.

Affiliations

¹ "Medical and Pharmaceutical Bionanotechnologies" Laboratory, Institute of Cellular Biology and Pathology "Nicolae Simionescu" of the Romanian Academy, 050568, Bucharest, Romania.
² "Liquid and Gas Chromatography" Laboratory, Institute of Cellular Biology and Pathology "Nicolae Simionescu" of the Romanian Academy, 050568, Bucharest, Romania.
³ Centre of Advanced Research in Bionanoconjugates and Biopolymers, "Petru Poni" Institute of Macromolecular Chemistry, 700487, Iasi, Romania.
⁴ Advanced Centre for Research-Development in Experimental Medicine, Grigore T. Popa University of Medicine and Pharmacy of Iasi, 700115, Iasi, Romania.
⁵ Central Military Hospital "Dr. Carol Davila", Cardiovascular Surgery Clinic, Bucharest, Romania.

Abstract

Runx2 is a key transcription factor involved in valvular interstitial cells (VIC) osteodifferentiation, a process actively entwined with the calcific aortic valve disease (CAVD). We hypothesize that a strategy intended to silence Runx2 could be a valuable novel therapeutic option for CAVD. To this intent, we aimed at (i) developing targeted nanoparticles for efficient delivery of short hairpin (sh)RNA sequences specific for Runx2 to the aortic valve employing a relevant mouse model for CAVD and (ii) investigate their therapeutic potential in osteoblast-differentiated VIC (oVIC) cultivated into a 3D scaffold. Since collagen IV was used as a target, a peptide that binds specifically to collagen IV (Cp) was conjugated to the surface of lipopolyplexes encapsulating shRNA-Runx2 (Cp-LPP/shRunx2). The results showed that Cp-LPP/shRunx2 were (i) cytocompatible; (ii) efficiently taken up by 3D-cultured oVIC; (iii) diminished the osteodifferentiation of human VIC (cultured in a 3D hydrogel-derived from native aortic root) by reducing osteogenic molecules expression, alkaline phosphatase activity, and calcium concentration; and (iv) were recruited in aortic valve leaflets in a murine model of atherosclerosis. Taken together, these data recommend Cp-LPP/shRunx2 as a novel targeted nanotherapy to block the progression of CAVD, with a good perspective to be introduced in practical use.

Keywords: Calcific aortic valve disease; Collagen IV; Lipopolyplexes; Runx2; Valvular interstitial cells; shRNA.