bims-novged Biomed News
on Non-viral vectors for gene delivery
Issue of 2023–06–04
nine papers selected by
the Merkel lab, Ludwig-Maximilians University
  1. Evaluation of a DoE based approach for comprehensive modelling of the effect of lipid nanoparticle composition on nucleic acid delivery.
  2. Studying how administration route and dose regulates antibody generation against LNPs for mRNA delivery with single-particle resolution.
  3. A Route to Synthesize Ionizable Lipid ALC-0315, a Key Component of the mRNA Vaccine Lipid Matrix.
  4. Low energy nebulization preserves integrity of SARS-CoV-2 mRNA vaccines for respiratory delivery.
  5. A modular RNA delivery system comprising spherical nucleic acids built on endosome-escaping polymeric nanoparticles.
  6. Dose-Dependent Production of Anti-PEG IgM after Intramuscular PEGylated-Hydrogenated Soy Phosphatidylcholine Liposomes, but Not Lipid Nanoparticle Formulations of DNA, Correlates with the Plasma Clearance of PEGylated Liposomal Doxorubicin in Rats.
  7. Lipid-based nanoparticle-mediated combination therapy for breast cancer management: a comprehensive review.
  8. Structural and componential design: new strategies regulating the behavior of lipid-based nanoparticles in vivo.
  9. PEG vs. zwitterions: How these surface decorations determine cellular uptake of lipid-based nanocarriers.
  1. Biomaterials. 2023 May 15. pii: S0142-9612(23)00166-7. [Epub ahead of print]299 122158
    Evaluation of a DoE based approach for comprehensive modelling of the effect of lipid nanoparticle composition on nucleic acid delivery.
    Yue Qin, Adam A Walters, Nadia Rouatbi, Julie Tzu-Wen Wang, Hend Mohamed Abdel-Bar, Khuloud T Al-Jamal.
      Therapeutic nucleic acids (TNAs) comprise an alternative to conventional drugs for cancer therapy. Recently, stable nucleic acid lipid particles (SNALPs) have been explored to deliver TNA efficiently and safely both in vitro and in vivo. Small interfering RNA (siRNA) and messenger RNA (mRNA) based drugs have been suggested for a wide range of pathologies, and their respective lipid nanoparticle (LNP) formulations have been optimised using a Design of Experiments (DoE) approach. However, it is uncertain as to whether data obtained from DoE using simple experimental outputs can be used to generate a general heuristic for delivery of diverse TNA both in vitro and in vivo. Using plasmid DNA (pDNA), for which limited DoE optimisation has been performed, and siRNA to represent the two extremities of the TNA spectrum in terms of size and biological requirements, we performed a comparative DoE for both molecules and assessed the predictive qualities of the model both in vitro and in vivo. By producing a minimum run of 24 SNALP formulations with different lipid compositions incorporating either pDNA or siRNA, DoE models were successfully established for predicting the effect of individual lipid composition on particle size, TNA encapsulation and transfection both in vitro and in vivo. The results showed that the particle size, and in vitro and in vivo transfection efficiency of both pDNA and siRNA SNALP formulations were affected by lipid compositions. The encapsulation efficiency of pDNA SNALPs but not siRNA SNALPs was affected by the lipid composition. Notably, the optimal lipid compositions of SNALPs for pDNA/siRNA delivery were not identical. Furthermore, in vitro transfection efficiency could not be used to predict promising LNP candidates in vivo. The DoE approach described in this study may provide a method for comprehensive optimisation of LNPs for various applications. The model and optimal formulation described in this study can serve as a foundation from which to develop other novel NA containing LNPs for multiple applications such as NA based vaccines, cancer immunotherapies and other TNA therapies.
    Keywords:  Cancer cells; Combinatory; Design of experiments; Lipid nanoparticles; Plasmid DNA; siRNA
    DOI:  https://doi.org/10.1016/j.biomaterials.2023.122158
  2. Mol Ther Methods Clin Dev. 2023 Jun 08. 29 450-459
    Studying how administration route and dose regulates antibody generation against LNPs for mRNA delivery with single-particle resolution.
    Rasmus Münter, Esben Christensen, Thomas L Andresen, Jannik B Larsen.
      Following the recent approval of both siRNA- and mRNA-based therapeutics, nucleic acid therapies are considered a game changer in medicine. Their envisioned widespread use for many therapeutic applications with an array of cellular target sites means that various administration routes will be employed. Concerns exist regarding adverse reactions against the lipid nanoparticles (LNPs) used for mRNA delivery, as PEG coatings on nanoparticles can induce severe antibody-mediated immune reactions, potentially being boosted by the inherently immunogenic nucleic acid cargo. While exhaustive information is available on how physicochemical features of nanoparticles affects immunogenicity, it remains unexplored how the fundamental choice of administration route regulates anti-particle immunity. Here, we directly compared antibody generation against PEGylated mRNA-carrying LNPs administered by the intravenous, intramuscular, or subcutaneous route, using a novel sophisticated assay capable of measuring antibody binding to authentic LNP surfaces with single-particle resolution. Intramuscular injections in mice were found to generate overall low and dose-independent levels of anti-LNP antibodies, while both intravenous and subcutaneous LNP injections generated substantial and highly dose-dependent levels. These findings demonstrate that before LNP-based mRNA medicines can be safely applied to new therapeutic applications, it will be crucial to carefully consider the choice of administration route.
    Keywords:  administration route; anti-drug antibodies; drug safety; fluorescence microscopy; lipid nanoparticle; nucleic acid delivery; poly(ethylene glycol); single-particle detection
    DOI:  https://doi.org/10.1016/j.omtm.2023.05.008
  3. Russ J Bioorg Chem. 2023 ;49(2): 412-415
    A Route to Synthesize Ionizable Lipid ALC-0315, a Key Component of the mRNA Vaccine Lipid Matrix.
    I A Boldyrev, V P Shendrikov, A G Vostrova, E L Vodovozova.
      The ionizable lipid ALC-0315, ((4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate), is a component of the lipid matrix of the prophylactic SARS-CoV-2 mRNA vaccine produced by Pfizer/BioNTech. This lipid ensures efficient vaccine assembly, protects the mRNA from premature degradation, and promotes the release of the nucleic acid into the cytoplasm for its further processing after endocytosis. The present work describes a simple and economical method for the synthesis of the ALC-0315 lipid, which can be taken advantage of in mRNA vaccine production.
    Keywords:  ALC-0315; ionizable lipid; lipid nanoparticles; mRNA; synthesis
    DOI:  https://doi.org/10.1134/S1068162023020061
  4. Sci Rep. 2023 May 31. 13(1): 8851
    Low energy nebulization preserves integrity of SARS-CoV-2 mRNA vaccines for respiratory delivery.
    Cees J M van Rijn, Killian E Vlaming, Reinout A Bem, Rob J Dekker, Albert Poortinga, Timo Breit, Selina van Leeuwen, Wim A Ensink, Kelly van Wijnbergen, John L van Hamme, Daniel Bonn, Teunis B H Geijtenbeek.
      Nebulization of mRNA therapeutics can be used to directly target the respiratory tract. A promising prospect is that mucosal administration of lipid nanoparticle (LNP)-based mRNA vaccines may lead to a more efficient protection against respiratory viruses. However, the nebulization process can rupture the LNP vehicles and degrade the mRNA molecules inside. Here we present a novel nebulization method able to preserve substantially the integrity of vaccines, as tested with two SARS-CoV-2 mRNA vaccines. We compare the new method with well-known nebulization methods used for medical respiratory applications. We find that a lower energy level in generating LNP droplets using the new nebulization method helps safeguard the integrity of the LNP and vaccine. By comparing nebulization techniques with different energy dissipation levels we find that LNPs and mRNAs can be kept largely intact if the energy dissipation remains below a threshold value, for LNP integrity 5-10 J/g and for mRNA integrity 10-20 J/g for both vaccines.
    DOI:  https://doi.org/10.1038/s41598-023-35872-4
  5. Nanoscale Adv. 2023 May 30. 5(11): 2941-2949
    A modular RNA delivery system comprising spherical nucleic acids built on endosome-escaping polymeric nanoparticles.
    Antonio Garcia-Guerra, Ruth Ellerington, Jens Gaitzsch, Jonathan Bath, Mahnseok Kye, Miguel A Varela, Giuseppe Battaglia, Matthew J A Wood, Raquel Manzano, Carlo Rinaldi, Andrew J Turberfield.
      Nucleic acid therapeutics require delivery systems to reach their targets. Key challenges to be overcome include avoidance of accumulation in cells of the mononuclear phagocyte system and escape from the endosomal pathway. Spherical nucleic acids (SNAs), in which a gold nanoparticle supports a corona of oligonucleotides, are promising carriers for nucleic acids with valuable properties including nuclease resistance, sequence-specific loading and control of receptor-mediated endocytosis. However, SNAs accumulate in the endosomal pathway and are thus vulnerable to lysosomal degradation or recycling exocytosis. Here, an alternative SNA core based on diblock copolymer PMPC25-PDPA72 is investigated. This pH-sensitive polymer self-assembles into vesicles with an intrinsic ability to escape endosomes via osmotic shock triggered by acidification-induced disassembly. DNA oligos conjugated to PMPC25-PDPA72 molecules form vesicles, or polymersomes, with DNA coronae on luminal and external surfaces. Nucleic acid cargoes or nucleic acid-tagged targeting moieties can be attached by hybridization to the coronal DNA. These polymeric SNAs are used to deliver siRNA duplexes against C9orf72, a genetic target with therapeutic potential for amyotrophic lateral sclerosis, to motor neuron-like cells. By attaching a neuron-specific targeting peptide to the PSNA corona, effective knock-down is achieved at doses of 2 particles per cell.
    DOI:  https://doi.org/10.1039/d2na00846g
  6. Mol Pharm. 2023 May 31.
    Dose-Dependent Production of Anti-PEG IgM after Intramuscular PEGylated-Hydrogenated Soy Phosphatidylcholine Liposomes, but Not Lipid Nanoparticle Formulations of DNA, Correlates with the Plasma Clearance of PEGylated Liposomal Doxorubicin in Rats.
    Christopher N Subasic, Neville J Butcher, Rodney F Minchin, Lisa M Kaminskas.
      PEGylated lipid nanoparticle-based Covid-19 vaccines, including Pfizer's BNT162b2 and Moderna's mRNA-1273, have been shown to stimulate variable anti-PEG antibody production in humans. Anti-PEG antibodies have the potential to accelerate the plasma clearance of PEGylated therapeutics, such as PEGylated liposomes and proteins, and compromise their therapeutic efficacy. However, it is not yet clear whether antibody titers produced by PEGylated Covid-19 vaccines significantly affect the clearance of PEGylated therapeutics. This study examined how anti-PEG IgM levels affect the pharmacokinetics of PEGylated liposomal doxorubicin (PLD) and compared the immunogenicity of a lipid nanoparticle formulation of linear DNA (DNA-LNP) to standard PEG-HSPC liposomes. DNA-LNP was prepared using the same composition and approach as Pfizer's BNT162b2, except linear double-stranded DNA was used as the genetic material. PEGylated HSPC-based liposomes were formulated using the lipid rehydration and extrusion method. Nanoparticles were dosed IM to rats at 0.005-0.5 mg lipid/kg body weight 1 week before evaluating the plasma pharmacokinetics of clinically relevant doses of PLD (1 mg/kg, IV) or PEGylated interferon α2a (Pegasys, 5 μg/kg, SC). Plasma PEG IgM was compared between pre- and 1-week post-dose blood samples. The results showed that anti-PEG IgM production increased with increasing PEG-HSPC liposome dose and that IgM significantly correlated with the plasma half-life, clearance, volume of distribution, and area under the curve of a subsequent dose of PLD. The plasma exposure of Pegasys was also significantly reduced after initial delivery of 0.005 mg/ml PEG-HSPC liposome. However, a single 0.05 mg/kg IM dose of DNA-LNP did not significantly elevate PEG IgM and did not alter the IV pharmacokinetics of PLD. These data showed that PEGylated Covid-19 vaccines are less immunogenic compared to standard PEGylated HSPC liposomes and that there is an antibody threshold for accelerating the clearance of PEGylated therapeutics.
    Keywords:  Covid-19; accelerated blood clearance; anti-PEG IgM; liposome; pharmacokinetics; poly(ethylene glycol)
    DOI:  https://doi.org/10.1021/acs.molpharmaceut.3c00104
  7. Drug Deliv Transl Res. 2023 Jun 01.
    Lipid-based nanoparticle-mediated combination therapy for breast cancer management: a comprehensive review.
    Priya Gupta, Yub Raj Neupane, Mohd Aqil, Kanchan Kohli, Yasmin Sultana.
      Breast cancer due to the unpredictable and complex etiopathology combined with the non-availability of any effective drug treatment has become the major root of concern for oncologists globally. The number of women affected by the said disease state is increasing at an alarming rate attributed to environmental and lifestyle changes indicating at the exploration of a novel treatment strategy that can eradicate this aggressive disease. So far, it is treated by promising nanomedicine monotherapy; however, according to the numerous studies conducted, the inadequacy of these nano monotherapies in terms of elevated toxicity and resistance has been reported. This review, therefore, puts forth a new multimodal strategic approach to lipid-based nanoparticle-mediated combination drug delivery in breast cancer, emphasizing the recent advancements. A basic overview about the combination therapy and its index is firstly given. Then, the various nano-based combinations of chemotherapeutics involving the combination delivery of synthetic and herbal agents are discussed along with their examples. Further, the recent exploration of chemotherapeutics co-delivery with small interfering RNA (siRNA) agents has also been explained herein. Finally, a section providing a brief description of the delivery of chemotherapeutic agents with monoclonal antibodies (mAbs) has been presented. From this review, we aim to provide the researchers with deep insight into the novel and much more effective combinational lipid-based nanoparticle-mediated nanomedicines tailored specifically for breast cancer treatment resulting in synergism, enhanced antitumor efficacy, and low toxic effects, subsequently overcoming the hurdles associated with conventional chemotherapy.
    Keywords:  Breast cancer; Combination index; Lipid-based nanoparticle-mediated combination therapy; Monoclonal antibodies; Nanomedicine monotherapy; Small interfering RNA; Synergism effect
    DOI:  https://doi.org/10.1007/s13346-023-01366-z
  8. Biomater Sci. 2023 May 30.
    Structural and componential design: new strategies regulating the behavior of lipid-based nanoparticles in vivo.
    Qingguo Zhong, Chunxiong Zheng, Ke Yi, Rachel L Mintz, Shixian Lv, Yu Tao, Mingqiang Li.
      Lipid-based nanoparticles have made a breakthrough in clinical disease as delivery systems due to their biocompatibility, thermal and long-term stability, high loading ability, simplicity of preparation, inexpensive production costs, and scalable manufacturing production. In particular, during the COVID-19 pandemic, this delivery system served as a vital vaccine component for virus confrontation. To obtain effective drug delivery, lipid-based nanoparticles should reach the desired sites with high efficiency, enter target cells, and release drugs. The structures and compositions of lipid-based nanoparticles can be modified to regulate these behaviors in vivo to enhance the therapeutic effects. Herein, we briefly review the development of lipid-based nanoparticles, from simple self-assembled nanovesicle-structured liposomes to multifunctional lipid nanoparticles. Subsequently, we summarize the strategies that regulate their tissue distribution, cell internalization, and drug release, highlighting the importance of the structural and componential design. We conclude with insights for further research to advance lipid-based nanotechnology.
    DOI:  https://doi.org/10.1039/d3bm00387f
  9. J Colloid Interface Sci. 2023 May 18. pii: S0021-9797(23)00870-6. [Epub ahead of print]647 52-64
    PEG vs. zwitterions: How these surface decorations determine cellular uptake of lipid-based nanocarriers.
    Daniel Stengel, Betül Hilal Demirel, Patrick Knoll, Martyna Truszkowska, Flavia Laffleur, Andreas Bernkop-Schnürch.
       AIM: To evaluate the impact of polyethylene glycol (PEG) and zwitterionic surface decoration of lipid-based nanocarriers (NC) on cellular uptake.
    METHODS: Anionic, neutral and cationic zwitterionic lipid-based NCs based on lecithin were compared with conventional PEGylated lipid-based NCs regarding stability in biorelevant fluids, interaction with endosome mimicking membranes, cytocompatibility, cellular uptake and permeation across intestinal mucosa.
    RESULTS: PEGylated and zwitterionic lipid-based NCs exhibited a droplet size between 100 and 125 nm with a narrow size distribution. For the PEGylated and zwitterionic lipid-based NCs only minor alterations in size and PDI in fasted state intestinal fluid and mucus containing buffer were observed, demonstrating similar bioinert properties. Erythrocytes interaction studies revealed enhanced endosomal escape properties for zwitterionic lipid-based NCs compared to PEGylated lipid-based NCs. For the zwitterionic lipid-based NCs negligible cytotoxicity on Caco-2 and HEK cells, even in the highest tested concentration of 1 % (v/v) was recorded. The PEGylated lipid-based NCs showed a cell survival of ≥75 % for concentrations ≤0.05 % on Caco-2 and HEK cells, which was considered as non-toxic. For the zwitterionic lipid-based NCs up to 60-fold higher cellular uptake on Caco-2 cells was determined compared to PEGylated lipid-based NCs. For the cationic zwitterionic lipid-based NCs the highest cellular uptake with 58.5 % and 40.0 % in Caco-2 and HEK cells, respectively, was determined. The results were confirmed visually by life cell imaging. Ex-vivo permeation experiments using rat intestinal mucosa demonstrated up to 8.6-fold enhanced permeation of the lipophilic marker coumarin-6 in zwitterionic lipid-based NCs compared to the control. Up to 6.9-fold enhanced permeation of coumarin-6 in neutral zwitterionic lipid-based NCs compared to the PEGylated counterpart was recorded.
    CONCLUSION: The replacement of PEG surfactants with zwitterionic surfactants is a promising approach to overcome the drawbacks of conventional PEGylated lipid-based NCs regarding intracellular drug delivery.
    Keywords:  Cellular uptake; Endosomal escape; Life cell imaging; Lipid-based nanocarrier; PEGylated surfactant; Zwitterionic surfactant
    DOI:  https://doi.org/10.1016/j.jcis.2023.05.079
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