Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles
Abstract Background Messenger RNA (mRNA) has gained remarkable attention as an alternative to DNA-based therapies in biomedical research. A variety of biodegradable nanoparticles (NPs) has been developed including lipid-based and polymer-based systems for mRNA delivery. However, both systems still l...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-09-01
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| Series: | Journal of Nanobiotechnology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12951-018-0401-y |
| _version_ | 1811301196336463872 |
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| author | Hanzey Yasar Alexander Biehl Chiara De Rossi Marcus Koch Xabi Murgia Brigitta Loretz Claus-Michael Lehr |
| author_facet | Hanzey Yasar Alexander Biehl Chiara De Rossi Marcus Koch Xabi Murgia Brigitta Loretz Claus-Michael Lehr |
| author_sort | Hanzey Yasar |
| collection | DOAJ |
| description | Abstract Background Messenger RNA (mRNA) has gained remarkable attention as an alternative to DNA-based therapies in biomedical research. A variety of biodegradable nanoparticles (NPs) has been developed including lipid-based and polymer-based systems for mRNA delivery. However, both systems still lack in achieving an efficient transfection rate and a detailed understanding of the mRNA transgene expression kinetics. Therefore, quantitative analysis of the time-dependent translation behavior would provide a better understanding of mRNA’s transient nature and further aid the enhancement of appropriate carriers with the perspective to generate future precision nanomedicines with quick response to treat various diseases. Results A lipid–polymer hybrid system complexed with mRNA was evaluated regarding its efficiency to transfect dendritic cells (DCs) by simultaneous live cell video imaging of both particle uptake and reporter gene expression. We prepared and optimized NPs consisting of poly (lactid-co-glycolid) (PLGA) coated with the cationic lipid 1, 2-di-O-octadecenyl-3-trimethylammonium propane abbreviated as LPNs. An earlier developed polymer-based delivery system (chitosan-PLGA NPs) served for comparison. Both NPs types were complexed with mRNA-mCherry at various ratios. While cellular uptake and toxicity of either NPs was comparable, LPNs showed a significantly higher transfection efficiency of ~ 80% while chitosan-PLGA NPs revealed only ~ 5%. Further kinetic analysis elicited a start of protein translation after 1 h, with a maximum after 4 h and drop of transgene expression after 48 h post-transfection, in agreement with the transient nature of mRNA. Conclusions Charge-mediated complexation of mRNA to NPs enables efficient and fast cellular delivery and subsequent protein translation. While cellular uptake of both NP types was comparable, mRNA transgene expression was superior to polymer-based NPs when delivered by lipid–polymer NPs. |
| first_indexed | 2024-04-13T07:04:35Z |
| format | Article |
| id | doaj.art-a1679658956f434ea859878d25c8507f |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-13T07:04:35Z |
| publishDate | 2018-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-a1679658956f434ea859878d25c8507f2022-12-22T02:57:02ZengBMCJournal of Nanobiotechnology1477-31552018-09-0116111910.1186/s12951-018-0401-yKinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticlesHanzey Yasar0Alexander Biehl1Chiara De Rossi2Marcus Koch3Xabi Murgia4Brigitta Loretz5Claus-Michael Lehr6Department of Drug Delivery (DDEL), Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI)Department of Drug Delivery (DDEL), Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI)Department of Drug Delivery (DDEL), Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI)INM-Leibniz Institute for New MaterialsDepartment of Drug Delivery (DDEL), Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI)Department of Drug Delivery (DDEL), Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI)Department of Drug Delivery (DDEL), Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI)Abstract Background Messenger RNA (mRNA) has gained remarkable attention as an alternative to DNA-based therapies in biomedical research. A variety of biodegradable nanoparticles (NPs) has been developed including lipid-based and polymer-based systems for mRNA delivery. However, both systems still lack in achieving an efficient transfection rate and a detailed understanding of the mRNA transgene expression kinetics. Therefore, quantitative analysis of the time-dependent translation behavior would provide a better understanding of mRNA’s transient nature and further aid the enhancement of appropriate carriers with the perspective to generate future precision nanomedicines with quick response to treat various diseases. Results A lipid–polymer hybrid system complexed with mRNA was evaluated regarding its efficiency to transfect dendritic cells (DCs) by simultaneous live cell video imaging of both particle uptake and reporter gene expression. We prepared and optimized NPs consisting of poly (lactid-co-glycolid) (PLGA) coated with the cationic lipid 1, 2-di-O-octadecenyl-3-trimethylammonium propane abbreviated as LPNs. An earlier developed polymer-based delivery system (chitosan-PLGA NPs) served for comparison. Both NPs types were complexed with mRNA-mCherry at various ratios. While cellular uptake and toxicity of either NPs was comparable, LPNs showed a significantly higher transfection efficiency of ~ 80% while chitosan-PLGA NPs revealed only ~ 5%. Further kinetic analysis elicited a start of protein translation after 1 h, with a maximum after 4 h and drop of transgene expression after 48 h post-transfection, in agreement with the transient nature of mRNA. Conclusions Charge-mediated complexation of mRNA to NPs enables efficient and fast cellular delivery and subsequent protein translation. While cellular uptake of both NP types was comparable, mRNA transgene expression was superior to polymer-based NPs when delivered by lipid–polymer NPs.http://link.springer.com/article/10.1186/s12951-018-0401-ymRNATransfectionGene deliveryChitosan-PLGACationic lipidLive cell imaging |
| spellingShingle | Hanzey Yasar Alexander Biehl Chiara De Rossi Marcus Koch Xabi Murgia Brigitta Loretz Claus-Michael Lehr Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles Journal of Nanobiotechnology mRNA Transfection Gene delivery Chitosan-PLGA Cationic lipid Live cell imaging |
| title | Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles |
| title_full | Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles |
| title_fullStr | Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles |
| title_full_unstemmed | Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles |
| title_short | Kinetics of mRNA delivery and protein translation in dendritic cells using lipid-coated PLGA nanoparticles |
| title_sort | kinetics of mrna delivery and protein translation in dendritic cells using lipid coated plga nanoparticles |
| topic | mRNA Transfection Gene delivery Chitosan-PLGA Cationic lipid Live cell imaging |
| url | http://link.springer.com/article/10.1186/s12951-018-0401-y |
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