Optimizing Cardiac Delivery of Modified mRNA.

Abstract

Modified mRNA (modRNA) is a new technology in the field of somatic gene transfer that has been used for the delivery of genes into different tissues, including the heart. Our group and others have shown that modRNAs injected into the heart are robustly translated into the encoded protein and can potentially improve outcome in heart injury models. However, the optimal compositions of the modRNA and the reagents necessary to achieve optimal expression in the heart have not been characterized yet. In this study, our aim was to elucidate those parameters by testing different nucleotide modifications, modRNA doses, and transfection reagents both in vitro and in vivo in cardiac cells and tissue. Our results indicate that optimal cardiac delivery of modRNA is with N1-Methylpseudouridine-5'-Triphosphate nucleotide modification and achieved using 0.013 μg modRNA/mm2/500 cardiomyocytes (CMs) transfected with positively charged transfection reagent in vitro and 100 μg/mouse heart (1.6 μg modRNA/μL in 60 μL total) sucrose-citrate buffer in vivo. We have optimized the conditions for cardiac delivery of modRNA in vitro and in vivo. Using the described methods and conditions may allow for successful gene delivery using modRNA in various models of cardiovascular disease.

• Authors: Nishat Sultana, Ajit Magadum, Yoav Hadas, Jason Kondrat, Neha Singh, Elias Youssef, Damelys Calderon, Elena Chepurko, Nicole Dubois, Roger J Hajjar, Lior Zangi
• Journal: Molecular therapy : the journal of the American Society of Gene Therapy (Mol Ther) Vol. 25 Issue 6 Pg. 1306-1315 (06 07 2017) ISSN: 1525-0024 [Electronic] United States
• PMID: 28389322 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.

Chemical References

• Biomarkers
• RNA, Messenger

Topics

• Animals
• Biomarkers
• Cells, Cultured
• Female
• Gene Expression
• Gene Transfer Techniques
• Genes, Reporter
• Humans
• Male
• Mice
• Molecular Imaging
• Myocardium (metabolism)
• Myocytes, Cardiac (metabolism)
• Protein Biosynthesis
• RNA, Messenger (administration & dosage, chemistry, genetics)
• Rats
• Transfection