Payload distribution and capacity of mRNA lipid nanoparticles

Li, Sixuan; Hu, Yizong; Li, Andrew; Lin, Jiaping; Hsieh, Kuangwen; Schneiderman, Zachary; Zhang, Pengfei; Zhu, Yining; Qiu, Chenhu; Kokkoli, Efrosini; Wang, Tza‐Huei; Mao, Hai‐Quan

doi:10.1038/s41467-022-33157-4

Cited by 108 publications

(79 citation statements)

References 28 publications

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“…The zeta potential at the low mass ratio for OC2-K3-E10 and SM-102 was more negative, while MC3 did not change significantly (Figure D). We observed large fluctuations in size across mass ratios for OC2-K3-E10 but not for SM-102 or MC3, hinting at differences in self-assembly dynamics between these ionizable lipids (Figure C). − Together, size, encapsulation, and zeta potential measurements suggest that low mass ratio LNPs are more loosely packed and that the mRNA is more accessible near the surface.…”

Section: Resultsmentioning

confidence: 95%

Iterative Design of Ionizable Lipids for Intramuscular mRNA Delivery

et al. 2023

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Lipid nanoparticles (LNPs) are the most clinically advanced delivery vehicles for RNA and have enabled the development of RNA-based drugs such as the mRNA COVID-19 vaccines. Functional delivery of mRNA by an LNP greatly depends on the inclusion of an ionizable lipid, and small changes to these lipid structures can significantly improve delivery. However, the structure–function relationships between ionizable lipids and mRNA delivery are poorly understood, especially for LNPs administered intramuscularly. Here, we show that the iterative design of a novel series of ionizable lipids generates key structure–activity relationships and enables the optimization of chemically distinct lipids with efficacy that is on-par with the current state of the art. We find that the combination of ionizable lipids comprising an ethanolamine core and LNPs with an apparent pKa between 6.6 and 6.9 maximizes intramuscular mRNA delivery. Furthermore, we report a nonlinear relationship between the lipid-to-mRNA mass ratio and protein expression, suggesting that a critical mass ratio exists for LNPs and may depend on ionizable lipid structure. Our findings add to the mechanistic understanding of ionizable lipids and demonstrate that hydrogen bonding, ionization behavior, and lipid-to-mRNA mass ratio are key design parameters affecting intramuscular mRNA delivery. We validate these insights by applying them to the rational design of new ionizable lipids. Overall, our iterative design strategy efficiently generates potent ionizable lipids. This hypothesis-driven method reveals structure–activity relationships that lay the foundation for the optimization of ionizable lipids in future LNP-RNA drugs. We foresee that this design strategy can be extended to other optimization parameters beyond intramuscular expression.

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Section: Resultsmentioning

confidence: 95%

Iterative Design of Ionizable Lipids for Intramuscular mRNA Delivery

et al. 2023

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“…The inclusion of PEG lipids is known to affect both the zeta potential and size of the resulting particle, increasing the stability of the formations by inhibiting their aggregation [ 85 ]. The percentage of PEG utilized as well as PEG conjugations also play a role in the characteristics of the LNPs [ 85 , 86 ]. For example, a larger PEG percentage can lead to the formation of smaller LNPs [ 86 ].…”

Section: Vehicles For Delivering Mrna To the Cellsmentioning

confidence: 99%

“…The percentage of PEG utilized as well as PEG conjugations also play a role in the characteristics of the LNPs [ 85 , 86 ]. For example, a larger PEG percentage can lead to the formation of smaller LNPs [ 86 ]. The conjugation of 1,2-dimyristoyl-rac-glycero-3-methoxy, which carries a C14 alkyl chain, can also modify behavior, providing lower circulation time in the bloodstream coupled with higher cellular delivery rates, compared to conjugates utilizing C18 1,2-distearoyl-rac-glycero-3-methoxy [ 87 ].…”

Section: Vehicles For Delivering Mrna To the Cellsmentioning

confidence: 99%

mRNA in the Context of Protein Replacement Therapy

et al. 2023

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Protein replacement therapy is an umbrella term used for medical treatments that aim to substitute or replenish specific protein deficiencies that result either from the protein being absent or non-functional due to mutations in affected patients. Traditionally, such an approach requires a well characterized but arduous and expensive protein production procedure that employs in vitro expression and translation of the pharmaceutical protein in host cells, followed by extensive purification steps. In the wake of the SARS-CoV-2 pandemic, mRNA-based pharmaceuticals were recruited to achieve rapid in vivo production of antigens, proving that the in vivo translation of exogenously administered mRNA is nowadays a viable therapeutic option. In addition, the urgency of the situation and worldwide demand for mRNA-based medicine has led to an evolution in relevant technologies, such as in vitro transcription and nanolipid carriers. In this review, we present preclinical and clinical applications of mRNA as a tool for protein replacement therapy, alongside with information pertaining to the manufacture of modified mRNA through in vitro transcription, carriers employed for its intracellular delivery and critical quality attributes pertaining to the finished product.

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“…Intraserum stability [ 149 ], anti-RNase performance, and storage stability [ 150 ] were also evaluated to ensure a stable product quality. The payload distribution and capacity of mRNA-LNPs are critical but remain a challenge, and some researchers have developed a method based on multi-laser cylindrical illumination confocal spectroscopy (CICS) [ 151 ].…”

Section: Mrna Delivery Systemmentioning

confidence: 99%

mRNA-Based Therapeutics in Cancer Treatment

et al. 2023

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Over the past two decades, significant technological innovations have led to messenger RNA (mRNA) becoming a promising option for developing prophylactic and therapeutic vaccines, protein replacement therapies, and genome engineering. The success of the two COVID-19 mRNA vaccines has sparked new enthusiasm for other medical applications, particularly in cancer treatment. In vitro-transcribed (IVT) mRNAs are structurally designed to resemble naturally occurring mature mRNA. Delivery of IVT mRNA via delivery platforms such as lipid nanoparticles allows host cells to produce many copies of encoded proteins, which can serve as antigens to stimulate immune responses or as additional beneficial proteins for supplements. mRNA-based cancer therapeutics include mRNA cancer vaccines, mRNA encoding cytokines, chimeric antigen receptors, tumor suppressors, and other combination therapies. To better understand the current development and research status of mRNA therapies for cancer treatment, this review focused on the molecular design, delivery systems, and clinical indications of mRNA therapies in cancer.

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Payload distribution and capacity of mRNA lipid nanoparticles

Cited by 108 publications

References 28 publications

Iterative Design of Ionizable Lipids for Intramuscular mRNA Delivery

Iterative Design of Ionizable Lipids for Intramuscular mRNA Delivery

mRNA in the Context of Protein Replacement Therapy

mRNA-Based Therapeutics in Cancer Treatment

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