Species-dependent in vivo mRNA delivery and cellular responses to nanoparticles

Hatit, Marine Z. C.; Lokugamage, Melissa P.; Dobrowolski, Curtis; Paunovska, Kalina; Ni, Huanzhen; Zhao, Kun; Vanover, Daryll; Beyersdorf, Jared; Peck, Hannah E.; Loughrey, David; Sato, Manaka; Cristian, Ana; Santangelo, Philip J.; Dahlman, James E.

doi:10.1038/s41565-021-01030-y

Cited by 74 publications

(62 citation statements)

References 42 publications

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“…However, it is difficult to compare in vitro and in vivo delivery 29 . One key limitation of this work is that LNP delivery may vary across species 52 , and thus, these results need to be confirmed in larger animals. Taken together, we believe the data justify further exploration of LNPs with piperazine rings.…”

Section: Discussionmentioning

confidence: 94%

Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo

Hatit

Zhao

et al. 2022

Nat Commun

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In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after intramuscular injection. However, systemic RNA delivery to non-hepatocytes remains challenging, especially without targeting ligands such as antibodies, peptides, or aptamers. Here we report that piperazine-containing ionizable lipids (Pi-Lipids) preferentially deliver mRNA to immune cells in vivo without targeting ligands. After synthesizing and characterizing Pi-Lipids, we use high-throughput DNA barcoding to quantify how 65 chemically distinct LNPs functionally delivered mRNA (i.e., mRNA translated into functional, gene-editing protein) in 14 cell types directly in vivo. By analyzing the relationships between lipid structure and cellular targeting, we identify lipid traits that increase delivery in vivo. In addition, we characterize Pi-A10, an LNP that preferentially delivers mRNA to the liver and splenic immune cells at the clinically relevant dose of 0.3 mg/kg. These data demonstrate that high-throughput in vivo studies can identify nanoparticles with natural non-hepatocyte tropism and support the hypothesis that lipids with bioactive small-molecule motifs can deliver mRNA in vivo.

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Section: Discussionmentioning

confidence: 94%

Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo

Hatit

Zhao

et al. 2022

Nat Commun

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“…The percentage of transfected DC is lower (40–70%) compared to that of HEK cells (60–80%). Lower transfection efficiency in murine dendritic cells was expected as they are notoriously harder to transfect than human cancer or immortalized cells [ 52 , 53 ] but it could also be attributed to the species-dependent cellular response to mRNA LNPs recently reported [ 54 ]. Whereas DOPE/Chol and DOPE/βS LNPs transfected 65–70% DCs—that is more than the commercial standard Lipofectamine Messenger Max ® (LFM, 45%)—DSPC/Chol LNPs only transfected 15% of DCs.…”

Section: Resultsmentioning

confidence: 99%

In Cellulo and In Vivo Comparison of Cholesterol, Beta-Sitosterol and Dioleylphosphatidylethanolamine for Lipid Nanoparticle Formulation of mRNA

et al. 2022

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Lipid Nanoparticles (LNPs) are a leading class of mRNA delivery systems. LNPs are made of an ionizable lipid, a polyethyleneglycol (PEG)-lipid conjugate and helper lipids. The success of LNPs is due to proprietary ionizable lipids and appropriate helper lipids. Using a benchmark lipid (D-Lin-MC3) we compared the ability of three helper lipids to transfect dendritic cells in cellulo and in vivo. Studies revealed that the choice of helper lipid does not influence the transfection efficiency of immortalized cells but, LNPs prepared with DOPE (dioleylphosphatidylethanolamine) and β-sitosterol were more efficient for mRNA transfection in murine dendritic cells than LNPs containing DSPC (distearoylphosphatidylcholine). This higher potency of DOPE and β-sitosterol LNPs for mRNA expression was also evident in vivo but only at low mRNA doses. Overall, these data provide valuable insight for the design of novel mRNA LNP vaccines.

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“…Mice and rats have larger liver compared to primates and this difference increase the off‐target liver delivery in murine models compared to human [23] . Species‐agnostic nanoparticle delivery screening (SANDS) has been developed to find the genes that differentiate the RNA delivery across species [23,250] . Using genetically engineered mouse with primatized or humanized liver can better predict the delivery of RNA and toxicity profile.…”

Section: Discussionmentioning

confidence: 99%

“…[23] Species-agnostic nanoparticle delivery screening (SANDS) has been developed to find the genes that differentiate the RNA delivery across species. [23,250] Using genetically engineered mouse with primatized or humanized liver can better predict the delivery of RNA and toxicity profile. More studies are needed to understand the mechanism of interaction of RNA cargo with the delivery vehicles and how it affects the delivery depending on cell state.…”

Section: Discussionmentioning

confidence: 99%

Emerging Approaches for Enabling RNAi Therapeutics

Mallick

Tripathi

Mishra

et al. 2022

Chemistry An Asian Journal

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RNA interference (RNAi) is a primitive evolutionary mechanism developed to escape incorporation of foreign genetic material. siRNA has been instrumental in achieving the therapeutic potential of RNAi by theoretically silencing any gene of interest in a reversible and sequence-specific manner. Extrinsically administered siRNA generally needs a delivery vehicle to span across different physiological barriers and load into the RISC complex in the cytoplasm in its functional form to show its efficacy. This review discusses the designing principles and examples of different classes of delivery vehicles that have proved to be efficient in RNAi therapeutics. We also briefly discuss the role of RNAi therapeutics in genetic and rare diseases, epigenetic modifications, immunomodulation and combination modality to inch closer in creating a personalized therapy for metastatic cancer. At the end, we present, strategies and look into the opportunities to develop efficient delivery vehicles for RNAi which can be translated into clinics.

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Species-dependent in vivo mRNA delivery and cellular responses to nanoparticles

Cited by 74 publications

References 42 publications

Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo

Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo

In Cellulo and In Vivo Comparison of Cholesterol, Beta-Sitosterol and Dioleylphosphatidylethanolamine for Lipid Nanoparticle Formulation of mRNA

Emerging Approaches for Enabling RNAi Therapeutics

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