Delivery of modified mRNA to damaged myocardium by systemic administration of lipid nanoparticles
Open accessAbstract
Lipid Nanoparticles (LNPs) are a promising drug delivery vehicle for clinical siRNA delivery. Modified mRNA (modRNA) has recently gained great attention as a therapeutic molecule in cardiac regeneration. However, for mRNA to be functional, it must first reach the diseased myocardium, enter the target cell, escape from the endosomal compartment into the cytosol and be translated into a functional protein. However, it is unknown if LNPs can effectively deliver mRNA, which is much larger than siRNA, to the ischemic myocardium. Here, we evaluated the ability of LNPs to deliver mRNA to the myocardium upon ischemia-reperfusion injury functionally. By exploring the bio-distribution of fluorescently labeled LNPs, we observed that, upon reperfusion, LNPs accumulated in the infarct area of the heart. Subsequently, the functional delivery of modRNA was evaluated by the administration of firefly luciferase encoding modRNA. Concomitantly, a significant increase in firefly luciferase expression was observed in the heart upon myocardial reperfusion when compared to sham-operated animals. To characterize the targeted cells within the myocardium, we injected LNPs loaded with Cre modRNA into Cre-reporter mice. Upon LNP infusion, Tdtomato+ cells, derived from Cre mediated recombination, were observed in the infarct region as well as the epicardial layer upon LNP infusion. Within the infarct area, most targeted cells were cardiac fibroblasts but also some cardiomyocytes and macrophages were found. Although the expression levels were low compared to LNP-modRNA delivery into the liver, our data show the ability of LNPs to functionally deliver modRNA therapeutics to the damaged myocardium, which holds great promise for modRNA-based cardiac therapies.
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Cas9 mRNA and sgRNA can be delivered into cells via electroporation or liposome-based vectors to improve genome editing fidelity.
“Similar to RNP electroporation, Cas9 mRNA and sgRNA can also be delivered into cells via electroporation or liposome-based vectors (Evers et al., 2022; Yu et al., 2023) to improved genome editing fidelity.”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as Evers et al., 2022
The Introduction and Discussion describe lipid-nanoparticle delivery of modified mRNA to damaged myocardium, and Methods §2.2 identifies the cargo as Cre mRNA. Searches for Cas9, sgRNA, electroporation, and genome-editing fidelity found no passage addressing the attributed claim; the general mRNA-delivery discussion does not establish it. (LLM, conf=0.76)
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Rodrigues, D. M. (2023) · doi:10.53846/goediss-9959
- Authors
- Evers, M. J. W., Du, W., Yang, Q., Kooijmans, S. A. A., Vink, A., van Steenbergen, M., Vader, P., de Jager, S. C. A., Fuchs, S. A., Mastrobattista, E., Sluijter, J. P. G., Lei, Z., & Schiffelers, R.
- Affiliations
- University Medical Center Utrecht (NL)
- Harbin Medical University (CN)
- Utrecht University (NL)
- Corresponding author
- Schiffelers, Raymond
- ORCID
- Venue
- Journal of Controlled Release
- Publisher
- Elsevier BV
- Published
- 2022-03
- Volume
- 343
- Pages
- 207-216
- ISSN
- 0168-3659 (ISSN-L)
- OpenAlex
- W4206915884
- Cited by
- 113
- References
- 48
- Citations by year
- 2026: 19 · 2025: 44 · 2024: 23 · 2023: 20 · 2022: 7
- FWCI
- 7.67
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Controlled Release (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Utrecht University Repository (Utrecht University) (submittedVersion, other-oa)
- Funders
- European Commission (HORIZON2020, 721058, 725229, 825828)
- ZonMw (725229, 116002016)
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (16169)
- Ministerie van Economische Zaken, Landbouw en Innovatie
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- lipid nanoparticles, modified mRNA, myocardial ischemia-reperfusion injury, cardiac regeneration, systemic drug delivery, firefly luciferase reporter, Cre reporter mice, cardiac fibroblasts
- MeSH terms
- Animals, Infarction, Liposomes, Myocardium, RNA, Messenger, RNA, Small Interfering, Luciferases, Firefly, Mice, Nanoparticles
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