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Translation, but not transfection limits clinically relevant, exogenous mRNA based induction of alpha-4 integrin expression on human mesenchymal stem cells
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Nowakowski, Adam, Andrzejewska, Anna, Boltze, Johannes, Nitzsche, Franziska, Cui, Li-li, Jolkkonen, Jukka, Walczak, Piotr, Lukomska, Barbara and Janowski, Miroslaw (2017) Translation, but not transfection limits clinically relevant, exogenous mRNA based induction of alpha-4 integrin expression on human mesenchymal stem cells. Scientific Reports, 7 (1). 1103 . doi:10.1038/s41598-017-01304-3
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WRAP-translation-exogenous-mRNA-based-induction-alpha-4-integrin-expression-human-mesenchymal-stem-cells-Boltze-2017.pdf - Published Version - Requires a PDF viewer. Available under License Creative Commons Attribution 4.0. Download (10Mb) | Preview |
Official URL: http://dx.doi.org/10.1038/s41598-017-01304-3
Abstract
Mesenchymal stem cells (MSCs) represent promising resource of cells for regenerative medicine in neurological disorders. However, efficient and minimally invasive methods of MSCs delivery to the brain still have to be developed. Intra-arterial route is very promising, but MSCs are missing machinery for diapedesis through blood-brain barrier. Thus, here we have tested a mRNA-based method to induce transient expression of ITGA4, an adhesion molecule actively involved in cell extravasation. We observed that transfection with an ITGA4-mRNA construct bearing a conventional cap analogue (7-methylguanosine) failed to produce ITGA4 protein, but exogenous ITGA4-mRNA was detected in transfected MSCs. This indicates that not transfection, but rather translation being the major roadblock. Stabilization of ITGA4-mRNA with SSB proteins resulted in ITGA4 protein synthesis in HEK293 cells only, whereas in MSCs, satisfactory results were obtained only after using an anti-reverse-cap-analogue (ARCA). The presence of ITGA4 protein in MSCs was transient and lasted for up to 24 h after transfection. Membranous location was confirmed by flow cytometry of viable non-permeabilized cells using anti-ITGA4 antibody. The mRNA-based expression of itga4 transgene is potentially sufficient for diapedesis after intra-arterial delivery. To conclude, mRNA-based engineering of stem cells is a rapid and integration-free method and attractive from the perspective of potential future clinical application.
| Item Type: | Journal Article | |||||||||
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| Subjects: | Q Science > QH Natural history > QH426 Genetics Q Science > QP Physiology |
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| Divisions: | Faculty of Science > Life Sciences (2010- ) | |||||||||
| Library of Congress Subject Headings (LCSH): | Mesenchymal stem cells, Messenger RNA | |||||||||
| Journal or Publication Title: | Scientific Reports | |||||||||
| Publisher: | Nature Publishing Group | |||||||||
| ISSN: | 2045-2322 | |||||||||
| Official Date: | 24 April 2017 | |||||||||
| Dates: |
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| Date of first compliant deposit: | 13 June 2019 | |||||||||
| Volume: | 7 | |||||||||
| Number: | 1 | |||||||||
| Article Number: | 1103 | |||||||||
| DOI: | 10.1038/s41598-017-01304-3 | |||||||||
| Status: | Peer Reviewed | |||||||||
| Publication Status: | Published | |||||||||
| Access rights to Published version: | Open Access | |||||||||
| RIOXX Funder/Project Grant: |
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