A Single Methylene Group in Oligoalkylamine‐Based Cationic Polymers and Lipids Promotes Enhanced mRNA Delivery

Jarzębińska, Anita; Pasewald, Tamara; Lambrecht, Jana; Mykhaylyk, Olga; Kümmerling, Linda; Beck, Philipp; Hasenpusch, Günther; Rudolph, Carsten; Plank, Christian; Dohmen, Christian

doi:10.1002/ange.201603648

Cited by 25 publications

(33 citation statements)

References 18 publications

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“…Cell-and tissue-specific delivery of cmRNA is currently considered a major obstacle to overcome for RTT. 26 We could show that intravenous delivery of reporter cmRNA in LLF as described by Jarzębi nska et al 35 led to strong and liver-specific protein translation ( Figures 4A and 4B). Immunohistochemical stainings indicate that intravenous application is a potent delivery mechanism for reaching a high number of hepatocytes, which was previously shown for delivery of small interfering RNA (siRNA) as well as mRNA.…”

Section: Discussionmentioning

confidence: 56%

“…Dysregulation of the local RAS contributes significantly to inflammation and fibrosis, 44,45 a process that can best be counterbalanced by specific ACE2 translation in the affected organs. For the purpose of liver-targeted cmRNA delivery, we chose lipoplexes (referred to as the liver lipidoid formulation [LLF] in the following) as described by Jarzębi nska et al 35 The liver specificity of LLF was first confirmed in a formulation with firefly luciferase cmRNA. A 1 mg/kg dose of this formulation was administered intravenously to mice and luciferase protein activity was measured 6 hr after administration.…”

Section: Ace2 Cmrna Can Be Selectively Targeted To the Liver And Lungmentioning

confidence: 99%

“…For liver-targeted in vivo experiments, cmRNA was formulated in LLF as previously described. 35 Lung-targeted lipoplexes (PLF) were prepared by mixing cmRNA with in-house polyethylene glycol (PEG) co-polymer in aqueous solution and applying it to a mix of preassembled lipid micelles. After incubation for 15 min at RT for self-assembly of cmRNA in this PLF, the mix was transferred into Dulbecco's PBS (Thermo Fisher Scientific) supplemented with 2% sucrose.…”

Section: Cmrna Preparationmentioning

confidence: 99%

“…This may be achieved with recent advances in cmRNA technology. [34][35][36] Therefore, the objective of this study was to establish robust ACE2 translation from cmRNA in the liver or lung, respectively. First, we performed an in vitro validation of ACE2 cmRNA translation, protein activity, and integrity.…”

Section: Introductionmentioning

confidence: 99%

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Translation of Angiotensin-Converting Enzyme 2 upon Liver- and Lung-Targeted Delivery of Optimized Chemically Modified mRNA

Schrom

Huber

Aneja

et al. 2017

Molecular Therapy - Nucleic Acids

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Changes in lifestyle and environmental conditions give rise to an increasing prevalence of liver and lung fibrosis, and both have a poor prognosis. Promising results have been reported for recombinant angiotensin-converting enzyme 2 (ACE2) protein administration in experimental liver and lung fibrosis. However, the full potential of ACE2 may be achieved by localized translation of a membrane-anchored form. For this purpose, we advanced the latest RNA technology for liver- and lung-targeted ACE2 translation. We demonstrated in vitro that transfection with ACE2 chemically modified messenger RNA (cmRNA) leads to robust translation of fully matured, membrane-anchored ACE2 protein. In a second step, we designed eight modified ACE2 cmRNA sequences and identified a lead sequence for in vivo application. Finally, formulation of this ACE2 cmRNA in tailor-made lipidoid nanoparticles and in lipid nanoparticles led to liver- and lung-targeted translation of significant amounts of ACE2 protein, respectively. In summary, we provide evidence that RNA transcript therapy (RTT) is a promising approach for ACE2-based treatment of liver and lung fibrosis to be tested in fibrotic disease models.

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Section: Discussionmentioning

confidence: 56%

Section: Ace2 Cmrna Can Be Selectively Targeted To the Liver And Lungmentioning

confidence: 99%

Section: Cmrna Preparationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Translation of Angiotensin-Converting Enzyme 2 upon Liver- and Lung-Targeted Delivery of Optimized Chemically Modified mRNA

Schrom

Huber

Aneja

et al. 2017

Molecular Therapy - Nucleic Acids

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“…Other research has also reported on the optimization of cation structure in polyplexes and lipoplexes, showing that properties of cations, such as charge density, charge number, and pH-dependent protonation largely determined the chemical and biological properties of the carriers, including nucleic acid compaction status, nuclease tolerability, and endosomal escaping capability. [49][50][51][52][53][54][55] The dissociation of nucleic acid carriers through interaction with GAGs or other molecules is another critical problem for in vivo nucleic acid delivery, and crosslinking the polymers or lipids comprising the carriers is an effective solution to this problem. However, after reaching the cytosol or nuclei of target cells, pDNA and mRNA in the carriers should be released to allow efficient transcription or translation.…”

Section: Introductionmentioning

confidence: 99%

Design concepts of polyplex micelles for in vivo therapeutic delivery of plasmid DNA and messenger RNA

Uchida

Kataoka

2019

J Biomedical Materials Res

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Nonviral delivery of plasmid (p)DNA or messenger (m)RNA is a safe and promising therapeutic option to continuously supply therapeutic proteins into diseased tissues. In most cases of in vivo pDNA and mRNA delivery, these nucleic acids are loaded into carriers based on cationic polymers and/or lipids to prevent nuclease‐mediated degradation before reaching target cells. The carriers should also evade host clearance mechanisms, including uptake by scavenger cells and filtration in the spleen. Installation of ligands onto the carriers can facilitate their rapid uptake into target cells. Meanwhile, carrier toxicity should be minimized not only for preventing undesirable adverse responses in patients, but also for preserving the function of transfected cells to exert therapeutic effects. Long‐term progressive improvement of platform technologies has helped overcome most of these issues, though some still remain hindering the widespread clinical application of nonviral pDNA and mRNA delivery. This review discusses design concepts of nonviral carriers for in vivo delivery and the issues to be overcome, focusing especially on our own efforts using polyplex micelles. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 978–990, 2019.

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Polymer–Lipid Nanoparticles for Systemic Delivery of mRNA to the Lungs

et al. 2016

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Therapeutic nucleic acids hold great promise for the treatment of disease but require vectors for safe and effective delivery. Synthetic nanoparticle vectors composed of poly(β-amino esters) (PBAEs) and nucleic acids have previously demonstrated potential utility for local delivery applications. To expand potential utility to include systemic delivery of mRNA, here we develop hybrid polymer-lipid nanoformulations for systemic delivery to the lung. Through co-formulation of PBAEs with lipid-polyethylene glycol (PEG), mRNA formulations were developed with increased serum stability and increased in vitro potency. The formulations were capable of functional delivery of mRNA to the lungs following intravenous administration in mice. To our knowledge, this is the first description of systemic administration of mRNA for delivery to the lungs using a degradable polymer-lipid nanoparticle. KeywordsNanoparticles; mRNA; Drug delivery; PBAE; Stability dgander@mit.edu. Supporting information for this article is given via a link at the end of the document.More detailed experimental procedures can be found in the supplementary information provided. Previous studies have suggested that serum stability may limit the efficacy of PBAEs for systemic administration. HHS Public Access[15] To this end, PBAE terpolymer copolymers containing internal alkyl tails were developed.[16] The alkyl amine was incorporated to enable formulation of terpolymers with other hydrophobic components in order to increase particle stability in physiological conditions. Here, we demonstrate that PBAE terpolymer nanoparticles can be formulated with a polyethylene glycol-modified lipid (PEG-lipid) and result in serum stabilized nanoparticles and improved efficacy. PBAEs that were found to be serum stabilized in vitro upon incorporation of PEG-lipid at 7 mol% were evaluated in vivo and shown to be capable of mRNA delivery to the lungs in mice following intravenous administration.The PBAEs selected for synthesis in this study were based on analysis of literature results from this class of materials for DNA delivery.[16] We synthesized 7 PBAEs that included 6 lead terpolymers and 1 lead non-terpolymer (i.e. not including the alkyl amine) from the monomers shown in Figure 1a. Synthetic methodology was adapted for synthesis in dimethylformamide (DMF) and purification in diethylether in order to reduce the possibility of toxicity arising from excess monomer (Scheme S1).[16] Polymer characterization was performed via GPC, 1H-NMR, and FTIR (Figures S1-3). Nanoparticles ( Figure 1b) were prepared by mixing each synthesized polymer with mRNA coding for firefly luciferase diluted in acidic (pH 5.2) buffer. Nanoparticles were formulated such that the ratio of amine groups in the polymer to phosphate groups in the nucleic acid (N/P) was 57; this is consistent with a weight ratio of approximately 50:1, similar to what has been used for successful PBAE transfections previously.[17]Stable nanoparticles were formed both with and without addition of 7 mol% PEG-lipid ...

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A Single Methylene Group in Oligoalkylamine‐Based Cationic Polymers and Lipids Promotes Enhanced mRNA Delivery

Abstract: Supportinginformation and the ORCID identification number(s) for the author(s) of this article can be found under http://dx. Communications 9591Angew.C hem. Int.E d. 2016, 55,9 591 -9595 2 016 Wiley-VCH VerlagG mbH &C o. KGaA,W einheim

Cited by 25 publications

References 18 publications

Translation of Angiotensin-Converting Enzyme 2 upon Liver- and Lung-Targeted Delivery of Optimized Chemically Modified mRNA

Translation of Angiotensin-Converting Enzyme 2 upon Liver- and Lung-Targeted Delivery of Optimized Chemically Modified mRNA

Design concepts of polyplex micelles for in vivo therapeutic delivery of plasmid DNA and messenger RNA

Polymer–Lipid Nanoparticles for Systemic Delivery of mRNA to the Lungs

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