bims-novged 2023-03-12 papers

bims-novged

Biomed News

on Non-viral vectors for gene delivery

Issue of 2023–03–12
seven papers selected by
the Merkel lab, Ludwig-Maximilians University

Shaping the future from the small scale: dry powder inhalation of CRISPR-Cas9 lipid nanoparticles for the treatment of lung diseases.
Delivering mRNA to a human NK cell line, NK-92 cells, by lipid nanoparticles.
Cation-free siRNA-cored nanocapsules for tumor-targeted RNAi therapy.
Visible Light-Conjugation with Triazolinediones as a Route to Degradable Poly(ethylene glycol)-Lipids for mRNA Lipid Nanoparticle Formulation.
A single-dose F1-based mRNA-LNP vaccine provides protection against the lethal plague bacterium.
Nanoparticle drug delivery systems for synergistic delivery of tumor therapy.
Chemistry, structure and function of approved oligonucleotide therapeutics.

Expert Opin Drug Deliv. 2023 Mar 10.

Shaping the future from the small scale: dry powder inhalation of CRISPR-Cas9 lipid nanoparticles for the treatment of lung diseases.

Simone P Carneiro, Antonietta Greco, Enrica Chiesa, Ida Genta, Olivia M Merkel.

   INTRODUCTION: Most lung diseases are serious conditions resulting from genetic and environmental causes associated with high mortality and severe symptoms. Currently, treatments available have a palliative effect and many targets are still considered undruggable. Gene therapy stands as an attractive approach to offering innovative therapeutic solutions. CRISPR-Cas9 has established a remarkable potential for genome editing with high selectivity to targeted mutations. To ensure high efficacy with minimum systemic exposure, the delivery and administration route are key components that must be investigated.
AREAS COVERED: This review is focused on the delivery of CRISPR-Cas9 to the lungs, taking advantage of lipid nanoparticles (LNPs), the most clinically advanced nucleic acid carriers. We also aim to highlight the benefits of pulmonary administration as a local delivery route and the use of spray drying to prepare stable nucleic acid-based dry powder formulations able to overcome multiple lung barriers.
EXPERT OPINION: Exploring the pulmonary administration to deliver CRISPR-Cas9 loaded in LNPs as a dry powder increases the chances to achieve high efficacy and reduced adverse effects. CRISPR-Cas9 loaded in LNP-embedded microparticles has not yet been reported in the literature but has the potential to reach and accumulate in target cells in the lung, thus, enhancing overall efficacy and safety.

Keywords:  CRISPR-Cas9; gene therapy; lipid nanoparticles; pulmonary administration; spray drying

DOI:  https://doi.org/10.1080/17425247.2023.2185220
Int J Pharm. 2023 Mar 08. pii: S0378-5173(23)00230-2. [Epub ahead of print] 122810

Delivering mRNA to a human NK cell line, NK-92 cells, by lipid nanoparticles.

Takashi Nakamura, Taisei Nakade, Yusuke Sato, Hideyoshi Harashima.

  In cancer immunotherapy, therapeutic methods targeting NK are highly expected. NK cell-based therapy using NK-92, a human NK cell line, has been clinically evaluated. Delivering mRNA into NK-92 cells is a potent strategy for enhancing its functions. However, the use of lipid nanoparticles (LNP) for this purpose has not yet been evaluated. We previously developed a LNP that was composed of CL1H6 (CL1H6-LNP) for the efficient delivery of siRNA to NK-92 cells, and the use of this material for delivering mRNA to NK-92 cells is reported in this study. Compared with a DLin-MC3-DMA based LNP, used as a benchmark, the CL1H6-LNP caused a high mRNA expression intensity and a cell transfection efficiency of 100%. The efficient mRNA delivery by this CL1H6-LNP is attributed to the high affinity for NK-92 cells and the intense, rapid fusion with the endosomal membrane. It therefore appears that the CL1H6-LNP could be a useful non-viral vector for modifying the NK-92 functions by mRNA. Our findings also provide some insights into the design and development of LNPs for delivering mRNA to NK-92 and NK cells.

Keywords:  Lipid nanoparticle; NK cell; NK-92; Non-viral; ionizable lipid; mRNA

DOI:  https://doi.org/10.1016/j.ijpharm.2023.122810
Acta Biomater. 2023 Mar 08. pii: S1742-7061(23)00129-0. [Epub ahead of print]

Cation-free siRNA-cored nanocapsules for tumor-targeted RNAi therapy.

Xinghua Huang, Jianwei Li, Guanyi Li, Binyu Ni, Ziji Liang, Haodong Chen, Chaozhang Xu, Jianhua Zhou, Jinsheng Huang, Shaohui Deng.

  Cation-associated cytotoxicity limits the systemic administration of RNA delivery in vivo, demanding the development of non-cationic nanosystems. In this study, cation-free polymer-siRNA nanocapsules with disulfide-crosslinked interlayer, namely T-SS(-), were prepared via the following steps: 1) complexation of siRNA with a cationic block polymer cRGD-poly(ethylene glycol)-b-poly[(2-aminoethanethiol)aspartamide]-b-poly{N'-[N-(2-aminoethyl)-2-ethylimino-1-aminomethyl]aspartamide}, abbreviated as cRGD-PEG-PAsp(MEA)-PAsp(C=N-DETA), 2) interlayer crosslinking via disulfide bond in pH 7.4 solution, and 3) removal of cationic DETA pendant at pH 5.0 via breakage of imide bond. The cationic-free nanocapsules with siRNA cores not only showed great performance (such as efficient siRNA encapsulation, high stability in serum, cancer cell targeting via cRGD modification, and GSH-triggered siRNA release), but also achieved tumor-targeted gene silencing in vivo. Moreover, the nanocapsules loaded with siRNA against polo-like kinase 1 (siRNA-PLK1) significantly inhibited tumor growth without showing cation-associated toxicity side effects and remarkably improved the survival rate of PC-3 tumor-bearing mice. The cation-free nanocapsules could potentially serve as a safe and effective platform for siRNA delivery. STATEMENT OF SIGNIFICANCE: Cation-associated toxicity limits the clinical translation of cationic carriers for siRNA delivery. Recently, several non-cationic carriers, such as siRNA micelles, DNA-based nanogels, and bottlebrush-architectured poly(ethylene glycol), have been developed to deliver siRNA. However, in these designs, siRNA as a hydrophilic macromolecule was attached to the nanoparticle surface instead of being encapsulated. Thus, it was easily degraded by serum nuclease and often induced immunogenicity. Herein, we demonstrate a new type of cation-free siRNA-cored polymeric nanocapsules. The developed nanocapsules not only showed capacities including efficient siRNA encapsulation, high stability in serum, and cancer cell targeting via cRGD modification, but also achieved an efficient tumor-targeted gene silencing in vivo. Importantly, unlike cationic carriers, the nanocapsules exhibited no cation-associated side effects.

Keywords:  GSH sensitivity; RNA delivery; cation-associated toxicity; cation-free; crosslinking

DOI:  https://doi.org/10.1016/j.actbio.2023.03.001
Angew Chem Int Ed Engl. 2023 Mar 10. e202301102

Visible Light-Conjugation with Triazolinediones as a Route to Degradable Poly(ethylene glycol)-Lipids for mRNA Lipid Nanoparticle Formulation.

Bianka Golba, Matthieu Soete, Zifu Zhong, Niek Sanders, Filip E Du Prez, Hannes A Houck, Bruno de Geest.

  Polyethylene glycol (PEG) is considered as the gold standard for colloidal stabilization of nanomedicines and prolonging the half-life of proteins. However, PEG is non-degradable and lacks functionality in the backbone, thereby limiting the scope of its potential. Herein, we introduced concomitantly functionality and degradability onto the PEG backbone by reacting PEG with 1,2,4-triazoline-3,5-diones (TADs) derivatives under visible light irradiation. The TAD-PEG conjugates were degradable in aqueous medium under physiological conditions, with the rate of hydrolysis depending on the pH and temperature of the medium. Next, a PEG-lipid was modified with TAD-derivatives. The obtained TAD-PEG-lipids could be successfully used for mRNA lipid nanoparticle (LNP) delivery and increased mRNA transfection efficiency on multiple cell cultures in vitro, without increasing cellular uptake of LNPs. In vivo, in mice, mRNA LNP formulation exhibited a similar tissue distribution as common LNPs, with a slight decrease in transfection efficiency. Our findings pave the road towards the design of degradable, backbone-functionalized PEG for applications in nanomedicine and beyond.

Keywords:  degradability, photoconjugation, triazolidinediones, poly(ethylene glycol), lipid nanoparticles

DOI:  https://doi.org/10.1002/anie.202301102
Sci Adv. 2023 Mar 10. 9(10): eadg1036

A single-dose F1-based mRNA-LNP vaccine provides protection against the lethal plague bacterium.

Edo Kon, Yinon Levy, Uri Elia, Hila Cohen, Inbal Hazan-Halevy, Moshe Aftalion, Assaf Ezra, Erez Bar-Haim, Gonna Somu Naidu, Yael Diesendruck, Shahar Rotem, Nitay Ad-El, Meir Goldsmith, Emanuelle Mamroud, Dan Peer, Ofer Cohen.

Messenger RNA (mRNA) lipid nanoparticle (LNP) vaccines have emerged as an effective vaccination strategy. Although currently applied toward viral pathogens, data concerning the platform's effectiveness against bacterial pathogens are limited. Here, we developed an effective mRNA-LNP vaccine against a lethal bacterial pathogen by optimizing mRNA payload guanine and cytosine content and antigen design. We designed a nucleoside-modified mRNA-LNP vaccine based on the bacterial F1 capsule antigen, a major protective component of Yersinia pestis, the etiological agent of plague. Plague is a rapidly deteriorating contagious disease that has killed millions of people during the history of humankind. Now, the disease is treated effectively with antibiotics; however, in the case of a multiple-antibiotic-resistant strain outbreak, alternative countermeasures are required. Our mRNA-LNP vaccine elicited humoral and cellular immunological responses in C57BL/6 mice and conferred rapid, full protection against lethal Y. pestis infection after a single dose. These data open avenues for urgently needed effective antibacterial vaccines.

DOI: https://doi.org/10.1126/sciadv.adg1036
Front Pharmacol. 2023 ;14 1111991

Nanoparticle drug delivery systems for synergistic delivery of tumor therapy.

Daoyuan Chen, Xuecun Liu, Xiaoyan Lu, Jingwei Tian.

  Nanoparticle drug delivery systems have proved anti-tumor effects; however, they are not widely used in tumor therapy due to insufficient ability to target specific sites, multidrug resistance to anti-tumor drugs, and the high toxicity of the drugs. With the development of RNAi technology, nucleic acids have been delivered to target sites to replace or correct defective genes or knock down specific genes. Also, synergistic therapeutic effects can be achieved for combined drug delivery, which is more effective for overcoming multidrug resistance of cancer cells. These combination therapies achieve better therapeutic effects than delivering nucleic acids or chemotherapeutic drugs alone, so the scope of combined drug delivery has also been expanded to three aspects: drug-drug, drug-gene, and gene-gene. This review summarizes the recent advances of nanocarriers to co-delivery agents, including i) the characterization and preparation of nanocarriers, such as lipid-based nanocarriers, polymer nanocarriers, and inorganic delivery carriers; ii) the advantages and disadvantages of synergistic delivery approaches; iii) the effectual delivery cases that are applied in the synergistic delivery systems; and iv) future perspectives in the design of nanoparticle drug delivery systems to co-deliver therapeutic agents.

Keywords:  co-delivery; combination therapy; nanoparticles; synergistic action; tumor therapy

DOI:  https://doi.org/10.3389/fphar.2023.1111991
Nucleic Acids Res. 2023 Mar 07. pii: gkad067. [Epub ahead of print]

Chemistry, structure and function of approved oligonucleotide therapeutics.

Martin Egli, Muthiah Manoharan.

Eighteen nucleic acid therapeutics have been approved for treatment of various diseases in the last 25 years. Their modes of action include antisense oligonucleotides (ASOs), splice-switching oligonucleotides (SSOs), RNA interference (RNAi) and an RNA aptamer against a protein. Among the diseases targeted by this new class of drugs are homozygous familial hypercholesterolemia, spinal muscular atrophy, Duchenne muscular dystrophy, hereditary transthyretin-mediated amyloidosis, familial chylomicronemia syndrome, acute hepatic porphyria, and primary hyperoxaluria. Chemical modification of DNA and RNA was central to making drugs out of oligonucleotides. Oligonucleotide therapeutics brought to market thus far contain just a handful of first- and second-generation modifications, among them 2'-fluoro-RNA, 2'-O-methyl RNA and the phosphorothioates that were introduced over 50 years ago. Two other privileged chemistries are 2'-O-(2-methoxyethyl)-RNA (MOE) and the phosphorodiamidate morpholinos (PMO). Given their importance in imparting oligonucleotides with high target affinity, metabolic stability and favorable pharmacokinetic and -dynamic properties, this article provides a review of these chemistries and their use in nucleic acid therapeutics. Breakthroughs in lipid formulation and GalNAc conjugation of modified oligonucleotides have paved the way to efficient delivery and robust, long-lasting silencing of genes. This review provides an account of the state-of-the-art of targeted oligo delivery to hepatocytes.

DOI: https://doi.org/10.1093/nar/gkad067