bims-novged 2023-04-16 papers

bims-novged

Biomed News

on Non-viral vectors for gene delivery

Issue of 2023–04–16
nine papers selected by
the Merkel lab, Ludwig-Maximilians University

Method of manufacturing CAF®09 liposomes affect immune responses induced by adjuvanted subunit proteins.
Core polymer optimization of ternary siRNA nanoparticles enhances in vivo safety, pharmacokinetics, and tumor gene silencing.
Nucleophile responsive charge-reversing polycations for pDNA transfection.
Inhaled Lipid Nanoparticles Alleviate Established Pulmonary Fibrosis.
Efficient mRNA Delivery with Lyophilized Human Serum Albumin-Based Nanobubbles.
Hyaluronic acid-conjugated liposomes loaded with dexamethasone: a promising approach for the treatment of inflammatory diseases.
Coarse-Grained Modeling of Polystyrene-Modified CNTs and Their Interactions with Lipid Bilayers.
A Multiple Targeting Nanoprobe for Identifying Cancer Metastatic Sites Based on Detection of Various mRNAs in Circulating Tumor Cells.
Meta-Analysis of Material Properties Influencing Nanoparticle Plasma Pharmacokinetics.

Eur J Pharm Biopharm. 2023 Apr 12. pii: S0939-6411(23)00085-1. [Epub ahead of print]

Method of manufacturing CAF®09 liposomes affect immune responses induced by adjuvanted subunit proteins.

Aneesh Thakur, Abishek Wadhwa, Abhijeet Lokras, Olivia Amanda Oest Müllertz, Dennis Christensen, Henrik Franzyk, Camilla Foged.

  The ability to induce antigen-specific CD4+ and CD8+ T-cell responses is one of the fundamental requirements when developing new efficacious vaccines against challenging infectious diseases and cancer. However, no adjuvants are currently approved for human subunit vaccines that induce T-cell immunity. Here, we incorporated a Toll-like receptor 4 agonist, i.e., the ionizable lipidoid L5N12, in the liposomal cationic adjuvant formulation 09 (CAF®09), and found that modified CAF®09 liposomes possess preserved adjuvant function as compared to unmodified CAF®09. CAF®09 consists of the cationic lipid dimethyldioctadecylammonium (DDA), monomycoloyl glycerol analogue 1 (MMG-1), and polyinosinic:polycytidylic acid [poly(I:C)]. By using the microfluidic mixing technology for liposome preparation, we gradually replaced DDA with L5N12, while keeping the molar ratios of MMG-1 and poly(I:C) constant. We found that this type of modification resulted in colloidally stable liposomes, which were significantly smaller and displayed reduced surface charge as compared to unmodified CAF®09, prepared by using the conventional thin film method. We showed that incorporation of L5N12 decreases the membrane rigidity of CAF®09 liposomes. Furthermore, vaccination with antigen adjuvanted with L5N12-modified CAF®09 or antigen adjuvanted with unmodified CAF®09, respectively, induced comparable antigen-specific serum antibody titers. We found that antigen adjuvanted with L5N12-modified CAF®09 induced antigen-specific effector and memory CD4+ and CD8+ T-cell responses in the spleen comparable to those induced when unmodified CAF®09 was used as adjuvant. However, incorporating L5N12 did not have a synergistic immunopotentiating effect on the antibody and T-cell responses induced by CAF®09. Moreover, vaccination with antigen adjuvanted with unmodified CAF®09, which was manufactured by using microfluidic mixing, induced significantly lower antigen-specific CD4+ and CD8+ T-cell responses than vaccination with antigen adjuvanted with unmodified CAF®09, which was prepared by using the thin film method. These results show that the method of manufacturing affects CAF®09 liposome adjuvanted antigen-specific immune responses, which should be taken into consideration when evaluating immunogenicity of subunit protein vaccines.

Keywords:  Lipidoid; TLR4 agonist; cationic adjuvant formulation 09 (CAF®09); drug delivery; liposomes; microfluidic mixing

DOI:  https://doi.org/10.1016/j.ejpb.2023.04.005
Biomaterials. 2023 Mar 28. pii: S0142-9612(23)00106-0. [Epub ahead of print]297 122098

Core polymer optimization of ternary siRNA nanoparticles enhances in vivo safety, pharmacokinetics, and tumor gene silencing.

Shrusti S Patel, Ella N Hoogenboezem, Fang Yu, Carlisle R DeJulius, R Brock Fletcher, Alex G Sorets, Fiona K Cherry, Justin H Lo, Mariah G Bezold, Nora Francini, Richard d'Arcy, Jordan E Brasuell, Rebecca S Cook, Craig L Duvall.

  Gene silencing with siRNA nanoparticles (si-NPs) is promising but still clinically unrealized for inhibition of tumor driver genes. Ternary si-NPs containing siRNA, a single block NP core-forming polymer poly[(2-(dimethylamino)ethyl methacrylate)-co-(butyl methacrylate)] (DMAEMA-co-BMA, 50B), and an NP surface-forming diblock polymer 20 kDa poly(ethylene glycol)-block-50B (20kPEG-50B) have the potential to improve silencing activity in tumors due to the participation of both 50B and 20kPEG-50B in siRNA electrostatic loading and endosome disruptive activity. Functionally, single block 50B provides more potent endosomolytic activity, while 20kPEG-50B colloidally stabilizes the si-NPs. Here, we systematically explored the role of the molecular weight (MW) of the core polymer and of the core:surface polymer ratio on ternary si-NP performance. A library of ternary si-NPs was formulated with variation in the MW of the 50B polymer and in the ratio of the core and surface forming polymeric components. Increasing 50B core polymer MW and ratio improved si-NP in vitro gene silencing potency, endosome disruptive activity, and stability, but these features also correlated with cytotoxicity. Concomitant optimization of 50B size and ratio resulted in the identification of lead ternary si-NPs 50B4-DP100, 50B8-DP100, and 50B12-DP25, with potent activity and minimal toxicity. Following intravenous treatment in vivo, all lead si-NPs displayed negligible toxicological effects and enhanced pharmacokinetics and tumor gene silencing relative to more canonical binary si-NPs. Critically, a single 1 mg/kg intravenous injection of 50B8-DP100 si-NPs silenced the tumor driver gene Rictor at the protein level by 80% in an orthotopic breast tumor model. 50B8-DP100 si-NPs delivering siRictor were assessed for therapeutic efficacy in an orthotopic HCC70 mammary tumor model. This formulation significantly inhibited tumor growth compared to siControl-NP treatment. 50B8-DP100 si-NPs were also evaluated for safety and were well-tolerated following a multi-dose treatment scheme. This work provides new insight on ternary si-NP structure-function relationships and identifies core polymer optimization strategies that can yield safe si-NP formulations with potent oncogene silencing.

Keywords:  Cancer; Drug delivery; RNA interference; Ternary nanoparticles

DOI:  https://doi.org/10.1016/j.biomaterials.2023.122098
Polym Chem. 2023 Apr 04. 14(14): 1591-1601

Nucleophile responsive charge-reversing polycations for pDNA transfection.

Reece W Lewis, Aswin Muralidharan, Benjamin Klemm, Pouyan E Boukany, Rienk Eelkema.

Polycationic carriers promise low cost and scalable gene therapy treatments, however inefficient intracellular unpacking of the genetic cargo has limited transfection efficiency. Charge-reversing polycations, which transition from cationic to neutral or negative charge, can offer targeted intracellular DNA release. We describe a new class of charge-reversing polycation which undergoes a cationic-to-neutral conversion by a reaction with cellular nucleophiles. The deionization reaction is relatively slow with primary amines, and much faster with thiols. In mammalian cells, the intracellular environment has elevated concentrations of amino acids (∼10×) and the thiol glutathione (∼1000×). We propose this allows for decationization of the polymeric carrier slowly in the extracellular space and then rapidly in the intracellular milleu for DNA release. We demonstrate that in a lipopolyplex formulation this leads to both improved transfection and reduced cytotoxicity when compared to a non-responsive polycationic control.

DOI: https://doi.org/10.1039/d3py00075c
Small. 2023 Apr 14. e2300545

Inhaled Lipid Nanoparticles Alleviate Established Pulmonary Fibrosis.

Dongjun Li, Ang Zhao, Jiafei Zhu, Chunjie Wang, Jingjing Shen, Zixuan Zheng, Feng Pan, Zhuang Liu, Qian Chen, Yang Yang.

  Pulmonary fibrosis, a sequela of lung injury resulting from severe infection such as severe acute respiratory syndrome-like coronavirus (SARS-CoV-2) infection, is a kind of life-threatening lung disease with limited therapeutic options. Herein, inhalable liposomes encapsulating metformin, a first-line antidiabetic drug that has been reported to effectively reverse pulmonary fibrosis by modulating multiple metabolic pathways, and nintedanib, a well-known antifibrotic drug that has been widely used in the clinic, are developed for pulmonary fibrosis treatment. The composition of liposomes made of neutral, cationic or anionic lipids, and poly(ethylene glycol) (PEG) is optimized by evaluating their retention in the lung after inhalation. Neutral liposomes with suitable PEG shielding are found to be ideal delivery carriers for metformin and nintedanib with significantly prolonged retention in the lung. Moreover, repeated noninvasive aerosol inhalation delivery of metformin and nintedanib loaded liposomes can effectively diminish the development of fibrosis and improve pulmonary function in bleomycin-induced pulmonary fibrosis by promoting myofibroblast deactivation and apoptosis, inhibiting transforming growth factor 1 (TGFβ1) action, suppressing collagen formation, and inducing lipogenic differentiation. Therefore, this work presents a versatile platform with promising clinical translation potential for the noninvasive inhalation delivery of drugs for respiratory disease treatment.

Keywords:  liposomes; metformin; nintedanib; pulmonary delivery; pulmonary fibrosis

DOI:  https://doi.org/10.1002/smll.202300545
Nanomaterials (Basel). 2023 Apr 05. pii: 1283. [Epub ahead of print]13(7):

Efficient mRNA Delivery with Lyophilized Human Serum Albumin-Based Nanobubbles.

Hiroshi Kida, Yutaro Yamasaki, Loreto B Feril, Hitomi Endo, Keiji Itaka, Katsuro Tachibana.

  In this study, we developed an efficient mRNA delivery vehicle by optimizing a lyophilization method for preserving human serum albumin-based nanobubbles (HSA-NBs), bypassing the need for artificial stabilizers. The morphology of the lyophilized material was verified using scanning electron microscopy, and the concentration, size, and mass of regenerated HSA-NBs were verified using flow cytometry, nanoparticle tracking analysis, and resonance mass measurements, and compared to those before lyophilization. The study also evaluated the response of HSA-NBs to 1 MHz ultrasound irradiation and their ultrasound (US) contrast effect. The functionality of the regenerated HSA-NBs was confirmed by an increased expression of intracellularly transferred Gluc mRNA, with increasing intensity of US irradiation. The results indicated that HSA-NBs retained their structural and functional integrity markedly, post-lyophilization. These findings support the potential of lyophilized HSA-NBs, as efficient imaging, and drug delivery systems for various medical applications.

Keywords:  drug delivery system; lyophilization; mRNA; nanobubble; sonoporation; ultrasound

DOI:  https://doi.org/10.3390/nano13071283
Int J Pharm. 2023 Apr 10. pii: S0378-5173(23)00366-6. [Epub ahead of print] 122946

Hyaluronic acid-conjugated liposomes loaded with dexamethasone: a promising approach for the treatment of inflammatory diseases.

Kamila B Japiassu, Francois Fay, Alessandro Marengo, Sebastião A Mendanha, Catherine Cailleau, Younès Louaguenouni, Qinglin Wang, Stéphanie Denis, Nicolas Tsapis, Thais L Nascimento, Eliana M Lima, Elias Fattal.

  Dexamethasone is a well-known anti-inflammatory drug readily used to treat many lung diseases. However, its side effects and poor lower airway deposition and retention are significant limitations to its usage. In this work, we developed lipid nanoparticulate platforms loaded with dexamethasone and evaluated their behavior in inflammatory lung models in vitro and in vivo. Dexamethasone-loaded liposomes with an average diameter below 150 nm were obtained using a solvent injection method. Three different formulations were produced with a distinct surface coating (polyethylene glycol, hyaluronic acid, or a mixture of both) as innovative strategies to cross the pulmonary mucus layer and/or target CD44 expressed on alveolar proinflammatory macrophages. Interestingly, while electron paramagnetic spectroscopy showed that surface modifications did not induce any molecular changes in the liposomal membrane, drug loading analysis revealed that adding the hyaluronic acid in the bilayer led to a decrease of dexamethasone loading (from 3.0 to 1.7w/w%). In vitro experiments on LPS-activated macrophages demonstrated that the encapsulation of dexamethasone in liposomes, particularly in HA-bearing ones, improved its anti-inflammatory efficacy compared to the free drug. Subsequently, in vivo data revealed that while intratracheal administration of free dexamethasone led to an important inter-animals variation of efficacy, dexamethasone-loaded liposomes showed an improved consistency within the results. Our data indicate that encapsulating dexamethasone into lipid nanoparticles is a potent strategy to improve its efficacy after lung delivery.

Keywords:  CD44 receptor; dexamethasone; inflammation; inhalation; liposomes

DOI:  https://doi.org/10.1016/j.ijpharm.2023.122946
Biophys J. 2023 Apr 12. pii: S0006-3495(23)00235-7. [Epub ahead of print]

Coarse-Grained Modeling of Polystyrene-Modified CNTs and Their Interactions with Lipid Bilayers.

Gulsah Gul, Roland Faller, Nazar Ileri-Ercan.

  In the present work, we describe Martini3 coarse-grained (CG) models of polystyrene (PS) and carboxyl-terminated polystyrene (PSCOOH) functionalized carbon nanotubes (CNTs) and investigate their interactions with lipid bilayers with and without cholesterol (CHOL) using molecular dynamics (MD) simulations. By changing the polystyrene chain length and grafting density at the end ring of the CNTs at two different nanotube concentrations, we observe the translocation of nanoparticles as well as changes in the lipid bilayer properties. Our results show that all developed models passively diffuse into the membranes without causing any damage to the membrane integrity although high concentrations of CNTs induce structural and elastic changes in lipid bilayers. In the presence of CHOL, increasing CNT concentration results in decreased rates of CHOL transmembrane motions. On the other hand, CNTs are prone to lipid and polystyrene blockage which affects their equilibrated configurations, and tilting behavior within the membranes. Hence, we demonstrate that polystyrene functionalized CNTs are promising drug-carrier agents. However, polystyrene chain length and grafting density are important factors to consider to enhance the efficiency of drug delivery.

Keywords:  Carbon nanotubes; lipid bilayer; molecular dynamics; polystyrene functionalization

DOI:  https://doi.org/10.1016/j.bpj.2023.04.005
Nano Lett. 2023 Apr 13.

A Multiple Targeting Nanoprobe for Identifying Cancer Metastatic Sites Based on Detection of Various mRNAs in Circulating Tumor Cells.

Di Han, Xiao-He Ren, Xin-Ru Liao, Xiao-Yan He, Tao Guo, Xue-Si Chen, Xuan Pang, Si-Xue Cheng.

  Identification of cancer metastatic sites is of importance for adjusting therapeutic interventions and treatment choice. However, identifying the location of metastatic lesions with easy accessibility and high safety is challenging. Here we demonstrate that cancer metastatic sites can be accurately detected by a triple targeting nanoprobe. Through coencapsulating molecular beacons probing a cancer biomarker (CXCR4 mRNA), a lung metastatic biomarker (CTSC mRNA), and a bone metastatic biomarker (JAG1 mRNA), the nanoprobe decorated by SYL3C conjugated hyaluronic acid and ICAM-1 specific aptamer conjugated hyaluronic acid can target diverse phenotyped circulating tumor cells (CTCs) during epithelial-mesenchymal and mesenchymal-epithelial transitions in whole blood for sensitive probing. The detection of CTCs from cancer patients shows that the nanoprobe can provide accurate information to distinguish different cancer metastasis statuses including nonmetastasis, lung metastasis, and bone metastasis. This study proposes an efficient screening tool for identifying the location of distant metastatic lesions via facile blood biopsy.

Keywords:  aptamer; cancer diagnosis; mRNA; metastasis; molecular beacon; targeting delivery

DOI:  https://doi.org/10.1021/acs.nanolett.2c04643
Int J Pharm. 2023 Apr 12. pii: S0378-5173(23)00371-X. [Epub ahead of print] 122951

Meta-Analysis of Material Properties Influencing Nanoparticle Plasma Pharmacokinetics.

Briana Macedo, Manthan Patel, Michael H Zaleski, Parth Mody, Xiaonan Ma, Patrick Mei, Jacob W Myerson, Jacob S Brenner, Patrick M Glassman.

  Thorough characterization of the plasma pharmacokinetics (PK) is a critical step in clinical development of novel therapeutics and is routinely performed for small molecules and biologics. However, there is a paucity of even basic characterization of PK for nanoparticle-based drug delivery systems. This has led to untested generalizations about how nanoparticle properties govern PK. Here, we present a meta-analysis of 100 nanoparticle formulations following IV administration in mice to identify any correlations between four PK parameters derived by non-compartmental analysis (NCA) and four cardinal properties of nanoparticles: PEGylation, zeta potential, size, and material. There was a statistically significant difference between the PK of particles stratified by nanoparticle properties. However, single linear regression between these properties and PK parameters showed poor predictability (r2 < 0.10 for all analyses), while multivariate regressions showed improved predictability (r2 > 0.38, except for t1/2). This suggests that no single nanoparticle property alone is even moderately predictive of PK, while the combination of multiple nanoparticle features does provide moderate predictive power. Improved reporting of nanoparticle properties will enable more accurate comparison between nanoformulations and will enhance our ability to predict in vivo behavior and design optimal nanoparticles.

Keywords:  Pharmacokinetics; meta-analysis; nanoparticles; non-compartmental analysis

DOI:  https://doi.org/10.1016/j.ijpharm.2023.122951