Lipoplexes for effective in vitro delivery of microRNAs to adult human cardiac fibroblasts for perspective direct cardiac cell reprogramming

• Published in: Nanomedicine Vol. 45; p. 102589
• Main Authors: Nicoletti, Letizia, Paoletti, Camilla, Tarricone, Giulia, Andreana, Ilaria, Stella, Barbara, Arpicco, Silvia, Divieto, Carla, Mattu, Clara, Chiono, Valeria
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01-09-2022
• Subjects: Adult human cardiac fibroblasts; Direct reprogramming; Lipoplexes; microRNAs; miRcombo; microRNAs; Adult human cardiac fibroblasts; Direct reprogramming; miRcombo; Lipoplexes
• ISSN: 1549-9634; 1549-9642
• DOI: 10.1016/j.nano.2022.102589

Design of nanocarriers for efficient miRNA delivery can significantly improve miRNA-based therapies. Lipoplexes based on helper lipid, dioleoyl phosphatidylethanolamine (DOPE) and cationic lipid [2-(2,3-didodecyloxypropyl)-hydroxyethyl] ammonium bromide (DE) were formulated to efficiently deliver miR-1 or a combination of four microRNAs (miRcombo) to adult human cardiac fibroblasts (AHCFs). Lipoplexes with amino-to-phosphate groups ratio of 3 (N/P 3) showed nanometric hydrodynamic size (372 nm), positive Z-potential (40 mV) and high stability under storage conditions. Compared to commercial DharmaFECT1 (DF), DE-DOPE/miRNA lipoplexes showed superior miRNA loading efficiency (99 % vs. 64 %), and faster miRNA release (99 % vs. 82 % at 48 h). DE-DOPE/miR-1 lipoplexes showed superior viability (80–100 % vs. 50 %) in AHCFs, a 2-fold higher miR-1 expression and Twinfilin-1 (TWF-1) mRNA downregulation. DE-DOPE/miRcombo lipoplexes significantly enhanced AHCFs reprogramming into induced cardiomyocytes (iCMs), as shown by increased expression of CM markers compared to DF/miRcombo.
We report the preparation and characterization of novel DE-DOPE/miRNA lipoplexes via simple electrostatic interactions, optimizing the ratio between the positively and negatively charged groups of the cationic lipid and miRNAs respectively. DE-DOPE/miRNA lipoplexes are able to efficiently load single miRNAs and miRNA combinations (miRcombo) and to fully release microRNA compared to commercial DharmaFECT1 (DF). Thus, DE-DOPE/miRcombo lipoplexes showed the ability to efficiently deliver miRcombo to AHCFs, with preliminary evidences of enhanced human fibroblast direct reprogramming into induced cardiomyocytes (iCMs). Image was created with Biorender under license. [Display omitted]
•Novel DE-DOPE/miRNA lipoplexes had high loading efficiency (99 %).•DE-DOPE/miRNA lipoplexes showed rapid and total release of microRNA.•Higher efficiency of DE-DOPE/miRNA lipoplexes for in vitro direct reprogramming.•DE-DOPE/miRNA lipoplexes were demonstrated to be optimal for in vitro miRNA release.