期刊
JOURNAL OF CONTROLLED RELEASE
卷 341, 期 -, 页码 206-214出版社
ELSEVIER
DOI: 10.1016/j.jconrel.2021.11.022
关键词
mRNA; Lipidoid; Lipid nanoparticles; Pseudouridine; Non-viral gene delivery
资金
- Defense Advanced Research Projects Agency (DARPA) [D16AP00143]
- Shurl and Kay Curci Foundation
- National Institutes of Health (NIH) [DP2-HD098860]
- NIH F32 Fellowship [1F32EB029345]
- NSF Graduate Research Fellowship Program award [DGE1745016]
- NIH (NIBIB) [R01 EB02519201A1]
- Cystic Fibrosis Foundation (CFF) [SIEGWA18XX0]
- Cancer Prevention and Research Institute of Texas (CPRIT) [RP190251]
- American Cancer Society (ACS) [RSG-17-012-01]
- NIAID [AI142596, AI124429]
- IPCAVD [U19AI142596]
Therapeutic mRNA shows potential in revolutionizing disease treatment, with base modifications impacting protein expression. Research found that m1 psi base modification significantly enhances translation, particularly with the most pronounced effects seen in the spleen.
Therapeutic mRNA has the potential to revolutionize the treatment of myriad diseases and, in 2020, facilitated the most rapid vaccine development in history. Among the substantial advances in mRNA technology made in recent years, the incorporation of base modifications into therapeutic mRNA sequences can reduce immunogenicity and increase translation. However, experiments from our lab and others have shown that the incorporation of base modifications does not always yield superior protein expression. We hypothesized that the variable benefit of base modifications may relate to lipid nanoparticle chemistry, formulation, and accumulation within specific organs. To test this theory, we compared IV-injected lipid nanoparticles formulated with reporter mRNA incorporating five base modifications (psi, m1 psi, m5U, m5C/psi, and m5C/s2U) and four ionizable lipids (C12-200, cKK-E12, ZA3-Ep10, and 200Oi10) with tropism for different organs. In general, the m1 psi base modification best enhanced translation, producing up to 15-fold improvements in total protein expression compared to unmodified mRNA. Expression improved most dramatically in the spleen (up to 50-fold) and was attributed to enhanced protein expression in monocytic lineage splenocytes. The extent to which these effects were observed varied with delivery vehicle and correlated with differences in innate immunogenicity. Through comparison of firefly luciferase and erythropoietin mRNA constructs, we also found that mRNA modification-induced enhancements in protein expression are limited outside of the spleen, irrespective of delivery vehicle. These results highlight the complexity of mRNA-loaded lipid nanoparticle drug design and show that the effectiveness of mRNA base modifications depend on the delivery vehicle, the target cells, and the site of endogenous protein expression.
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