bims-novged 2023-06-25 papers

bims-novged

Biomed News

on Non-viral vectors for gene delivery

Issue of 2023‒06‒25
ten papers selected by
the Merkel lab
Ludwig-Maximilians University

PEGylated pH-responsive peptide-mRNA nano self-assemblies enhance the pulmonary delivery efficiency and safety of aerosolized mRNA.
Engineering polyphenol-based carriers for nucleic acid delivery.
Rational design and combinatorial chemistry of ionizable lipids for RNA delivery.
The impact of nucleoside base modification in mRNA vaccine is influenced by the chemistry of its lipid nanoparticle delivery system.
Apoptosis-targeted gene therapy for non-small cell lung cancer using chitosan-poly-lactic-co-glycolic acid -based nano-delivery system and CASP8 and miRs 29A-B1 and 34A.
Genome Editing in Ferret Airway Epithelia Mediated by CRISPR/nucleases Delivered with Amphiphilic Shuttle Peptides.
Delivery systems of therapeutic nucleic acids for the treatment of acute lung injury/acute respiratory distress syndrome.
Novel α-mannose-functionalized poly(β-amino ester) nanoparticles as mRNA vaccines with increased antigen presenting cell selectivity in the spleen.
Dual-bioactive molecules loaded aligned core-shell microfibers for tendon tissue engineering.
Unravelling the role of lipid composition on liposome-protein interactions.

Drug Deliv. 2023 Dec;30(1): 2219870

PEGylated pH-responsive peptide-mRNA nano self-assemblies enhance the pulmonary delivery efficiency and safety of aerosolized mRNA.

Yingying Xu, Yijing Zheng, Xuqiu Ding, Chengyan Wang, Bin Hua, Shilian Hong, Xiaoman Huang, Jiali Lin, Peng Zhang, Wei Chen.

  Inhalable messenger RNA (mRNA) has demonstrated great potential in therapy and vaccine development to confront various lung diseases. However, few gene vectors could overcome the airway mucus and intracellular barriers for successful pulmonary mRNA delivery. Apart from the low pulmonary gene delivery efficiency, nonnegligible toxicity is another common problem that impedes the clinical application of many non-viral vectors. PEGylated cationic peptide-based mRNA delivery vector is a prospective approach to enhance the pulmonary delivery efficacy and safety of aerosolized mRNA by oral inhalation administration. In this study, different lengths of hydrophilic PEG chains were covalently linked to an amphiphilic, water-soluble pH-responsive peptide, and the peptide/mRNA nano self-assemblies were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The in vitro mRNA binding and release, cellular uptake, transfection, and cytotoxicity were studied, and finally, a proper PEGylated peptide with enhanced pulmonary mRNA delivery efficiency and improved safety in mice was identified. These results showed that a proper N-terminus PEGylation strategy using 12-monomer linear monodisperse PEG could significantly improve the mRNA transfection efficiency and biocompatibility of the non-PEGylated cationic peptide carrier, while a longer PEG chain modification adversely decreased the cellular uptake and transfection on A549 and HepG2 cells, emphasizing the importance of a proper PEG chain length selection. Moreover, the optimized PEGylated peptide showed a significantly enhanced mRNA pulmonary delivery efficiency and ameliorated safety profiles over the non-PEGylated peptide and LipofectamineTM 2000 in mice. Our results reveal that the PEGylated peptide could be a promising mRNA delivery vector candidate for inhaled mRNA vaccines and therapeutic applications for the prevention and treatment of different respiratory diseases in the future.

Keywords:  PEGylated peptide gene vector; Pulmonary mRNA delivery; inhalation safety; nano self-assemblies; pH-responsive peptide

DOI:  https://doi.org/10.1080/10717544.2023.2219870
Theranostics. 2023 ;13(10): 3204-3223

Engineering polyphenol-based carriers for nucleic acid delivery.

Mingju Shui, Zhejie Chen, Yi Chen, Qin Yuan, Hongyi Li, Chi Teng Vong, Mohamed A Farag, Shengpeng Wang.

  Gene therapy, an effective medical intervention strategy, is increasingly employed in basic research and clinical practice for promising and unique therapeutic effects for diseases treatment, such as cardiovascular disorders, cancer, neurological pathologies, infectious diseases, and wound healing. However, naked DNA/RNA is readily hydrolyzed by nucleic acid degrading enzymes in the extracellular environment and degraded by lysosomes during intracellular physiological conditions, thus gene transfer must cross complex cellular and tissue barriers to deliver genetic materials into targeted cells and drive efficient activation or inhibition of the proteins. At present, the lack of safe, highly efficient, and non-immunogenic drug carriers is the main drawback of gene therapy. Considering the dense hydroxyl groups on the benzene rings in natural polyphenols that exert a strong affinity to various nucleic acids via hydrogen bonding and hydrophobic interactions, polyphenol-based carriers are promising anchors for gene delivery in which polyphenols serve as the primary building blocks. In this review, the recent progress in polyphenol-assisted gene delivery was summarized, which provided an easily accessible reference for the design of future polyphenol-based gene delivery vectors. Nucleic acids discussed in this review include DNA, short interfering RNAs (siRNA), microRNA (miRNA), double-strand RNA (dsRNA), and messenger RNA (mRNA).

Keywords:  EGCG; RNA; gene therapy; nucleic acid; polyphenols

DOI:  https://doi.org/10.7150/thno.81604
J Mater Chem B. 2023 Jun 22.

Rational design and combinatorial chemistry of ionizable lipids for RNA delivery.

Yue Xu, Alex Golubovic, Shufen Xu, Anni Pan, Bowen Li.

In 2018, LNPs enabled the first FDA approval of a siRNA drug (Onpattro); two years later, two SARS-CoV-2 vaccines (Comirnaty, Spikevax) based on LNPs containing mRNA also arrived at the clinic, saving millions of lives during the COVID-19 pandemic. Notably, each of the three FDA-approved LNP formulations uses a unique ionizable lipid while the other three components, i.e., cholesterol, helper lipid, and PEGylated lipid, are almost identical. Therefore, ionizable lipids are critical to the delivery efficiency of mRNA. This review covers recent advances in ionizable lipids used in RNA delivery over the past several decades. We will discuss chemical structures, synthetic routes, and structure-activity relationships of ionizable lipids.

DOI: https://doi.org/10.1039/d3tb00649b
Mol Ther Nucleic Acids. 2023 Jun 13. 32 794-806

The impact of nucleoside base modification in mRNA vaccine is influenced by the chemistry of its lipid nanoparticle delivery system.

Marie-Clotilde Bernard, Emilie Bazin, Nadine Petiot, Katia Lemdani, Sylvie Commandeur, Cécile Verdelet, Sylvie Margot, Vladimir Perkov, Manon Ripoll, Marie Garinot, Sophie Ruiz, Florence Boudet, Bachra Rokbi, Jean Haensler.

  The use of modified nucleosides is an important approach to mitigate the intrinsic immunostimulatory activity of exogenous mRNA and to increase its translation for mRNA therapeutic applications. However, for vaccine applications, the intrinsic immunostimulatory nature of unmodified mRNA could help induce productive immunity. Additionally, the ionizable lipid nanoparticles (LNPs) used to deliver mRNA vaccines can possess immunostimulatory properties that may influence the impact of nucleoside modification. Here we show that uridine replacement with N1-methylpseudouridine in an mRNA vaccine encoding influenza hemagglutinin had a significant impact on the induction of innate chemokines/cytokines and a positive impact on the induction of functional antibody titers in mice and macaques when MC3 or KC2 LNPs were used as delivery systems, while it impacted only minimally the titers obtained with L319 LNPs, indicating that the impact of nucleoside modification on mRNA vaccine efficacy varies with LNP composition. In line with previous observations, we noticed an inverse correlation between the induction of high innate IFN-α titers in the macaques and antigen-specific immune responses. Furthermore, and consistent with the species specificity of pathogen recognition receptors, we found that the effect of uridine replacement did not strictly translate from mice to non-human primates.

Keywords:  MT: Oligonucleotides: Therapies and Applications; N1-methylpseudouridine; delivery system; immune response; innate immunity; ionizable lipids; lipid nanoparticle; mRNA modification; mRNA vaccine

DOI:  https://doi.org/10.1016/j.omtn.2023.05.004
Front Bioeng Biotechnol. 2023 ;11 1188652

Apoptosis-targeted gene therapy for non-small cell lung cancer using chitosan-poly-lactic-co-glycolic acid -based nano-delivery system and CASP8 and miRs 29A-B1 and 34A.

Sourav Chattopadhyay, Shashanka Shekhar Sarkar, Sheetanshu Saproo, Sheetal Yadav, Deepika Antil, Bodhisatwa Das, Srivatsava Naidu.

  Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related deaths worldwide, with resistance to apoptosis being a major driver of therapeutic resistance and aggressive phenotype. This study aimed to develop a novel gene therapy approach for NSCLC by targeting resistance to apoptosis. Loss of function mutations of caspase 8 (CASP8) and downregulation of microRNAs (miRs) 29A-B1 and 34A were identified as key contributors to resistance to apoptosis in NSCLC. A biodegradable polymeric nano-gene delivery system composed of chitosan-poly-lactic-co-glycolic acid was formulated to deliver initiator CASP8 and miRs 29A-B1 and 34A. The nano-formulation efficiently encapsulated the therapeutic genes effectively internalized into NSCLC cells and induced significant apoptosis. Evaluation of the nano-formulation in A549 tumor spheroids showed a significant increase in apoptosis within the core of the spheroids, suggesting effective penetration into the spheroid structures. We provide a novel nano-formulation that demonstrate therapeutic potential for suicidal gene therapy in NSCLC.

Keywords:  MicroRNAs; combinatorial gene therapy; gene delivery; nano-formulations; non-small cell lung cancer; tumor spheroids

DOI:  https://doi.org/10.3389/fbioe.2023.1188652
Hum Gene Ther. 2023 Jun 19.

Genome Editing in Ferret Airway Epithelia Mediated by CRISPR/nucleases Delivered with Amphiphilic Shuttle Peptides.

Meihui Luo, Jia Ma, Xue Cheng, Shuang Wu, Douglas J Bartels, David Guay, John F Engelhardt, Xiaoming Liu.

Gene editing strategies are attractive for treating genetic pulmonary diseases such as cystic fibrosis (CF). However, challenges have included the development of safe and effective vector systems for gene editing of airway epithelia and model systems to report their efficiency and durability. The domestic ferret (Mustela putorius furo) has a high degree of conservation in lung cellular anatomy with humans, and has served as an excellent model for many types of lung diseases including the CF. Here, we evaluated the efficiency of amphiphilic shuttle peptide S10 for protein delivery and gene editing using SpCas9, and AsCas12a (Cpf1) ribonucleoproteins (RNPs). These approaches were evaluated in proliferating ferret airway basal cells, polarized airway epithelia in vitro, and lungs in vivo, by accessing the editing efficiency using reporter ferrets and measuring indels at the ferret CFTR locus. Our results demonstrate that shuttle peptides efficiently enable delivery of reporter proteins/peptides and gene editing SpCas9 or Cpf1 RNP complexes to ferret airway epithelial cells in vitro and in vivo. We measured S10 delivery efficiency of GFP-NLS protein or SpCas9 RNP into ferret airway basal cells and fully differentiated ciliated and non-ciliated epithelial cells in vitro. In vitro and in vivo gene editing efficiencies were determined by Cas/LoxP-gRNA RNP-mediated conversion of a ROSA-TG Cre recombinase reporter using transgenic primary cells and ferrets. S10/Cas9 RNP was more effective relative to S10/Cpf1 RNP at gene editing of the ROSA-TG locus. Intratracheal lung delivery of the S10 shuttle combined with GFP-NLS protein or DRI-NLS peptide demonstrated efficiencies of protein delivery that were ~3-fold or 14-fold greater, respectively, than the efficiency of gene editing at the ROSA-TG locus using S10/Cas9/LoxP-gRNA. Cpf1 RNPs was less effective than SpCas9 at gene editing of LoxP locus. These data demonstrate the feasibility of shuttle peptide delivery of Cas RNPs to the ferret airways and the potential utility for developing ex vivo stem cell-based and in vivo gene editing therapies for genetic pulmonary diseases such as CF.

DOI: https://doi.org/10.1089/hum.2023.016
J Control Release. 2023 Jun 19. pii: S0168-3659(23)00391-7. [Epub ahead of print]360 1-14

Delivery systems of therapeutic nucleic acids for the treatment of acute lung injury/acute respiratory distress syndrome.

Chuanyu Zhuang, Minji Kang, Minhyung Lee.

  Acute lung injury (ALI)/ acute respiratory distress syndrome (ARDS) is a devastating inflammatory lung disease with a high mortality rate. ALI/ARDS is induced by various causes, including sepsis, infections, thoracic trauma, and inhalation of toxic reagents. Corona virus infection disease-19 (COVID-19) is also a major cause of ALI/ARDS. ALI/ARDS is characterized by inflammatory injury and increased vascular permeability, resulting in lung edema and hypoxemia. Currently available treatments for ALI/ARDS are limited, but do include mechanical ventilation for gas exchange and treatments supportive of reduction of severe symptoms. Anti-inflammatory drugs such as corticosteroids have been suggested, but their clinical effects are controversial with possible side-effects. Therefore, novel treatment modalities have been developed for ALI/ARDS, including therapeutic nucleic acids. Two classes of therapeutic nucleic acids are in use. The first constitutes knock-in genes for encoding therapeutic proteins such as heme oxygenase-1 (HO-1) and adiponectin (APN) at the site of disease. The other is oligonucleotides such as small interfering RNAs and antisense oligonucleotides for knock-down expression of target genes. Carriers have been developed for efficient delivery for therapeutic nucleic acids into the lungs based on the characteristics of the nucleic acids, administration routes, and targeting cells. In this review, ALI/ARDS gene therapy is discussed mainly in terms of delivery systems. The pathophysiology of ALI/ARDS, therapeutic genes, and their delivery strategies are presented for development of ALI/ARDS gene therapy. The current progress suggests that selected and appropriate delivery systems of therapeutic nucleic acids into the lungs may be useful for the treatment of ALI/ARDS.

Keywords:  Acute lung injury; Acute respiratory distress syndrome; Gene delivery; Gene therapy; Therapeutic nucleic acids

DOI:  https://doi.org/10.1016/j.jconrel.2023.06.018
J Mater Chem B. 2023 Jun 23.

Novel α-mannose-functionalized poly(β-amino ester) nanoparticles as mRNA vaccines with increased antigen presenting cell selectivity in the spleen.

Nil González-Ríos, Margalida Artigues, Marta Guerra-Rebollo, Antoni Planas, Salvador Borrós, Magda Faijes, Cristina Fornaguera.

mRNA vaccination has emerged as a prominent therapy for the future of medicine. Despite the colossal advance in this technology and worldwide efficacy proof (ca. COVID vaccines), mRNA carriers still lack cell/tissue specificity, leading to possible side effects, and reduced efficacy among others. Herein we make use of the ubiquitous affinity of antigen-presenting cells (APC)s for glycosides to achieve specific targeting. To achieve this goal, we designed a new generation of α-mannosyl functionalized oligopeptide-terminated poly(β-aminoester). Fine formulation of these polymers with mRNA resulted in nanoparticles decorated with surface-exposed α-mannoses with sizes around 180 nm and positive surface charge. Notably, these particles maintained their properties after freeze-drying and subsequent redispersion. Finally, our mRNA carriers preferentially targeted and transfected APCs in vitro and in vivo. In conclusion, we demonstrated, at a preclinical level, that the mannose functionalization enables more selective targeting of APCs and, thus, these polymer and nanoparticles are candidates for a new generation of mRNA immunotherapy vaccines.

DOI: https://doi.org/10.1039/d3tb00607g
Colloids Surf B Biointerfaces. 2023 Jun 17. pii: S0927-7765(23)00294-1. [Epub ahead of print]228 113416

Dual-bioactive molecules loaded aligned core-shell microfibers for tendon tissue engineering.

Hongyun Xuan, Zhuojun Zhang, Wei Jiang, Nianci Li, Li Sun, Ye Xue, Haitao Guan, Huihua Yuan.

  Development of a controlled delivery ultrafine fibrous system with two bioactive molecules is required to stimulate tendon healing in different phase. In this study, we used emulsion stable jet electrospinning to fabricate aligned poly(L-lactic acid) (PLLA) based ultrafine fibers with two small bioactive molecules of L-Arginine (Arg) and low molecular weight hyaluronic acid (HA). The results demonstrated that the aligned Arg/HA/PLLA microfibrous scaffold showed core-shell structure and allowed sequential release of Arg and HA due to their different electric charge. The scaffold also showed enhanced hydrophilicity, cell migration, spread and proliferation. Using an Achilles tendon repair model in rats, we demonstrated that this novel fibrous scaffold can prevent adhesion and promote tendon regeneration. Additionally, two p53 and ER-α-mediated signalling pathways were described as the probable main path of synergistic effects of the novel scaffold on tendon generation. Thus, this study may provide an important strategy for developing biofunctional and biomimetic tendon scaffolds.

Keywords:  Bioactive molecules; Controlled delivery ultrafine fibrous system; Tendon tissue engineering

DOI:  https://doi.org/10.1016/j.colsurfb.2023.113416
J Liposome Res. 2023 Jun 20. 1-9

Unravelling the role of lipid composition on liposome-protein interactions.

Valeria Nele, Federica D'Aria, Virginia Campani, Teresa Silvestri, Marco Biondi, Concetta Giancola, Giuseppe De Rosa.

  Upon in vivo administration of nanoparticles, a protein corona forms on their surface and affects their half-life in circulation, biodistribution properties, and stability; in turn, the composition of the protein corona depends on the physico-chemical properties of the nanoparticles. We have previously observed lipid composition-dependent in vitro and in vivo microRNA delivery from lipid nanoparticles. Here, we carried out an extensive physico-chemical characterisation to understand the role of the lipid composition on the in vivo fate of lipid-based nanoparticles. We used a combination of differential scanning calorimetry (DSC), membrane deformability measurements, isothermal titration calorimetry (ITC), and dynamic light scattering (DLS) to probe the interactions between the nanoparticle surface and bovine serum albumin (BSA) as a model protein. The lipid composition influenced membrane deformability, improved lipid intermixing, and affected the formation of lipid domains while BSA binding to the liposome surface was affected by the PEGylated lipid content and the presence of cholesterol. These findings highlight the importance of the lipid composition on the protein-liposome interaction and provide important insights for the design of lipid-based nanoparticles for drug delivery applications.

Keywords:  Lipid nanoparticles; differential scanning calorimetry; isothermal titration calorimetry; membrane deformability; protein corona

DOI:  https://doi.org/10.1080/08982104.2023.2224449