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

Letizia Nicoletti; Camilla Paoletti; Giulia Tarricone; Ilaria Andreana; Barbara Stella; Silvia Arpicco; Carla Divieto; Clara Mattu; Valeria Chiono

doi:10.1016/j.nano.2022.102589

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

Nanomedicine. 2022 Sep:45:102589. doi: 10.1016/j.nano.2022.102589. Epub 2022 Jul 28.

Authors

Letizia Nicoletti¹, Camilla Paoletti¹, Giulia Tarricone¹, Ilaria Andreana², Barbara Stella², Silvia Arpicco², Carla Divieto³, Clara Mattu¹, Valeria Chiono⁴

Affiliations

¹ Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy.
² Department of Drug Science and Technology, University of Turin, Via Pietro Giuria, 11, 10125, Turin, Italy.
³ Istituto Nazionale di Ricerca Metrologica, Division of Advanced Materials and Life Sciences, Str. delle Cacce, 91, 10135 Turin, Italy.
⁴ Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy. Electronic address: valeria.chiono@polito.it.

PMID: 35908737
DOI: 10.1016/j.nano.2022.102589

Abstract

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.

Keywords: Adult human cardiac fibroblasts; Direct reprogramming; Lipoplexes; miRcombo; microRNAs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cellular Reprogramming
Fibroblasts
Humans
Liposomes*
MicroRNAs* / genetics
Phosphates
Phosphatidylethanolamines
RNA, Messenger
Transfection

Substances

Liposomes
MicroRNAs
Phosphates
Phosphatidylethanolamines
RNA, Messenger