bims-novged Biomed News
on Non-viral vectors for gene delivery
Issue of 2022‒05‒22
thirteen papers selected by
Merkel Lab
Ludwig-Maximilians University
  1. Hydroxycholesterol substitution in Ionizable lipid nanoparticles for mRNA delivery to T cells.
  2. Current synopsis on siRNA therapeutics as a novel anti-cancer and antiviral strategy: Progress and challenges.
  3. Lipid-Nanoparticle-Based Delivery of CRISPR/Cas9 Genome-Editing Components.
  4. An imidazole modified lipid confers enhanced mRNA-LNP stability and strong immunization properties in mice and non-human primates.
  5. Glycopolymer-Cell-Penetrating Peptide (CPP) Conjugates for Efficient Epidermal Growth Factor Receptor (EGFR) Silencing.
  6. Multifunctional liposomal nanostructure-mediated siRNA/bortezomib co-delivery for SHARP1 knockdown in MLL-AF6 acute myeloid leukemia.
  7. Exciting Times for Lipid Nanoparticles: How Canadian Discoveries Are Enabling Gene Therapies.
  8. Structure-based peptide ligand design for improved epidermal growth factor receptor targeted gene delivery.
  9. PreS/2-21-Guided siRNA Nanoparticles Target to Inhibit Hepatitis B Virus Infection and Replication.
  10. RNA in cancer immunotherapy: unlocking the potential of the immune system.
  11. The paradigm shift in treatment from Covid-19 to oncology with mRNA vaccines.
  12. Albumin-based nanoparticles: a promising strategy to overcome cancer drug resistance.
  13. Nanoscale delivery platforms for RNA therapeutics: Challenges and the current state of the art.
  1. J Control Release. 2022 May 12. pii: S0168-3659(22)00282-6. [Epub ahead of print]
    Hydroxycholesterol substitution in Ionizable lipid nanoparticles for mRNA delivery to T cells.
    Savan K Patel, Margaret M Billingsley, Caitlin Frazee, Xuexiang Han, Kelsey L Swingle, Jingya Qin, Mohamad-Gabriel Alameh, Karin Wang, Drew Weissman, Michael J Mitchell.
      Delivery of nucleic acids, such as mRNA, to immune cells has become a major focus in the past decade with ionizable lipid nanoparticles (LNPs) emerging as a clinically-validated delivery platform. LNPs-typically composed of ionizable lipids, cholesterol, phospholipids, and polyethylene glycol-have been designed and optimized for a variety of applications including cancer therapies, vaccines, and gene editing. However, LNPs have only recently been investigated for delivery to T cells, which has various therapeutic applications including the engineering of T cell immunotherapies. While several LNP formulations have been evaluated for mRNA delivery, recent work has demonstrated that the utilization of cholesterol analogs may enhance mRNA delivery. Other studies have shown that cholesterols modified with hydroxyl groups can alter endocytic recycling mechanisms. Here, we engineered a library of LNPs incorporating hydroxycholesterols to evaluate their impact on mRNA delivery to T cells by leveraging endosomal trafficking mechanisms. Substitution of 25% and 50% 7α-hydroxycholesterol for cholesterol in LNPs enhanced mRNA delivery to primary human T cells ex vivo by 1.8-fold and 2.0-fold, respectively. Investigation of endosomal trafficking revealed that these modifications also increase late endosome production and reduce the presence of recycling endosomes. These results suggest that hydroxyl modification of cholesterol molecules incorporated into LNP formulations provides a mechanism for improving delivery of nucleic acid cargo to T cells for a range of immunotherapy applications.
    Keywords:  Cholesterol; Endosomal trafficking; Lipid nanoparticles; mRNA delivery
    DOI:  https://doi.org/10.1016/j.jconrel.2022.05.020
  2. Curr Pharm Biotechnol. 2022 May 16.
    Current synopsis on siRNA therapeutics as a novel anti-cancer and antiviral strategy: Progress and challenges.
    Sanjay Sharma, Rupesh K Gautam, Abhishek Kanugo, Dinesh Kumar Mishra, Mohammad Amjad Kamal.
      Currently, a popular era in nanomedicine is the implementation of RNA nanoparticles for various diseases and their diagnosis. RNA interference (RNAi) involves the arrangement of gene mediating mechanisms where coding and non-coding are carried out. The targeted control of gene utterance via siRNA system by nanocarriers showed an epic impact on modifying therapeutic efficacy. The article endeavours to highlight the mechanism of siRNA with concern to possible applications which are established on cancer therapy. In the current scenario to discuss the possible anti-viral effectiveness of nanoparticles with particular reference to SARS-CoV. Self-assembled nanoparticle (NP) is developed and it competently delivers to small interfering RNA (siRNA) intravenously to the tumour. The nanoparticle was found by mixing with siRNA, carrier, DNA, and lipids, preceded by after-change. Newly FDA appreciation of the first polymer-drug and additional ones in the clinically linked RNA polymer has to be highly therapeutic and diagnostic value. It has been established to be a particularly useful means for cell-type definite delivery of other RNA therapeutics like siRNA. While RNAi has helped speed up the discovery and understanding functions of a gene, it also has great potential as a therapeutic and potentially prophylactic modality. This article stated the development in the RNA polymer and also provides some examples of their diagnostic applications and therapeutics special emphasis on the anti-cancer and antiviral strategy. Patisiran and Givosiran are the recently approved si-RNA based products available in the market.
    Keywords:  Anti-viral; Cancer therapy; Drug Delivery; RNA; SARS; Treatment; siRNA Nanoparticle
    DOI:  https://doi.org/10.2174/1389201023666220516120432
  3. Mol Pharm. 2022 May 20.
    Lipid-Nanoparticle-Based Delivery of CRISPR/Cas9 Genome-Editing Components.
    Pardis Kazemian, Si-Yue Yu, Sarah B Thomson, Alexandra Birkenshaw, Blair R Leavitt, Colin J D Ross.
      Gene editing mediated by CRISPR/Cas9 systems is due to become a beneficial therapeutic option for treating genetic diseases and some cancers. However, there are challenges in delivering CRISPR components which necessitate sophisticated delivery systems for safe and effective genome editing. Lipid nanoparticles (LNPs) have become an attractive nonviral delivery platform for CRISPR-mediated genome editing due to their low immunogenicity and application flexibility. In this review, we provide a background of CRISPR-mediated gene therapy, as well as LNPs and their applicable characteristics for delivering CRISPR components. We then highlight the challenges of CRISPR delivery, which have driven the significant development of new, safe, and optimized LNP formulations in the past decade. Finally, we discuss considerations for using LNPs to deliver CRISPR and future perspectives on clinical translation of LNP-CRISPR gene editing.
    Keywords:  CRISPR/Cas9; gene therapy; genome editing; lipid nanoparticles; nanomedicine
    DOI:  https://doi.org/10.1021/acs.molpharmaceut.1c00916
  4. Biomaterials. 2022 May 07. pii: S0142-9612(22)00210-1. [Epub ahead of print]286 121570
    An imidazole modified lipid confers enhanced mRNA-LNP stability and strong immunization properties in mice and non-human primates.
    Manon Ripoll, Marie-Clotilde Bernard, Céline Vaure, Emilie Bazin, Sylvie Commandeur, Vladimir Perkov, Katia Lemdani, Marie-Claire Nicolaï, Patrick Bonifassi, Antoine Kichler, Benoit Frisch, Jean Haensler.
      The mRNA vaccine technology has promising applications to fight infectious diseases as demonstrated by the licensing of two mRNA-based vaccines, Comirnaty® (Pfizer/BioNtech) and Spikevax® (Moderna), in the context of the Covid-19 crisis. Safe and effective delivery systems are essential to the performance of these vaccines and lipid nanoparticles (LNPs) able to entrap, protect and deliver the mRNA in vivo are considered by many as the current "best in class". Nevertheless, current mRNA/LNP vaccine technology has still some limitations, one of them being thermostability, as evidenced by the ultracold distribution chain required for the licensed vaccines. We found that the thermostability of mRNA/LNP, could be improved by a novel imidazole modified lipid, DOG-IM4, in combination with standard helper lipids. DOG-IM4 comprises an ionizable head group consisting of imidazole, a dioleoyl lipid tail and a short flexible polyoxyethylene spacer between the head and tail. Here we describe the synthesis of DOG-IM4 and show that DOG-IM4 LNPs confer strong immunization properties to influenza HA mRNA in mice and macaques and a remarkable stability to the encapsulated mRNA when stored liquid in phosphate buffered saline at 4 °C. We speculate the increased stability to result from some specific attributes of the lipid's imidazole head group.
    Keywords:  Imidazole lipid; Ionizable lipid; Lipid nanoparticles; Storage stability; mRNA vaccine
    DOI:  https://doi.org/10.1016/j.biomaterials.2022.121570
  5. ACS Macro Lett. 2022 Apr 19. 11(4): 580-587
    Glycopolymer-Cell-Penetrating Peptide (CPP) Conjugates for Efficient Epidermal Growth Factor Receptor (EGFR) Silencing.
    Yi-Yang Peng, Haimei Hu, Diana Diaz-Dussan, Jianyang Zhao, Xiaojuan Hao, Ravin Narain.
      Overexpression of epidermal growth factor receptor (EGFR) is observed in multiple cancers such as colorectal, lung, and cervical solid tumors. Regulating the EGFR expression is an efficient strategy to manage these malignancies, and it can be achieved by using short interfering RNA (siRNA). Cell-penetrating peptides (CPPs) demonstrated an excellent capability to enhance the cellular uptake of siRNA, but high knockdown efficiencies have not been achieved due to endosomal entrapment. In this work, Schiff's base reaction was used to modify a block {P[LAEMA(2-lactobionamidoethyl methacrylamide)37]-b-P[FPMA(4-formyl phenyl methacrylate)2-st-DMA(N,N-dimethylacrylamide)2], P2} and two statistical [P(LAEMA23-st-FPMA3) (P3) and P(LAEMA25-st-FPMA2-st-DMA2) (P4)] aldehyde-based and galactose-based polymers, prepared via reversible addition-fragmentation chain-transfer (RAFT) polymerization. An arginine-rich peptide (ARP, KRRKRRRRRK) was used as a cell-penetrating peptide (CPP) and conjugated to the polymers via a Schiff base reaction. The resulting glycopolymer-peptide conjugates were utilized to condense the siRNA to prepare polyplexes with multivalent CPPs (MCPPs, a nanoparticle with multiple copies of the CPP) to enhance the endosomal escape. The polyplexes have different surface properties as determined by the architecture of polymers and the insertion of dimethyl amide moieties. The enhancement of cellular internalization of ARP was observed by labeling the polyplexes with fluorescein isothiocyanate (FITC)-siRNA showing a localization of polyplexes in the cytoplasm of a HeLa (cervical cancer) cell line. In the in vitro EFGR silencing study, the statistical glycopolymer-peptide (P3-P) polyplexes had superior EGFR silencing efficiency in comparison with the other polymers that were studied. Furthermore, P3-P polyplexes led to less off-targeting silencing than lipofectamine 3000. These encouraging results confirmed the potency of decorating galactose-based polymers with CPP, like ARP for their application in siRNA delivery and management of cervical carcinomas.
    DOI:  https://doi.org/10.1021/acsmacrolett.2c00046
  6. Mater Sci Eng C Mater Biol Appl. 2022 Jan 26. pii: S0928-4931(22)00023-6. [Epub ahead of print] 112663
    Multifunctional liposomal nanostructure-mediated siRNA/bortezomib co-delivery for SHARP1 knockdown in MLL-AF6 acute myeloid leukemia.
    Sameh A Mohammed, Yang Ju.
      Acute myeloid leukemia (AML) has an extremely poor prognosis and high relapse and fatality rates. New therapeutic mechanisms for molecular targeted delivery are urgently needed to improve patient survival. In this study, we targeted the oncogenic transcription factor SHARP1 using multifunctional small interfering RNA (siRNA) and bortezomib (BTZ)-loaded cRGD-guided PEGylated cationic liposomal nanostructures to monitor their antileukemic activity in MLL-AF6 AML cells. Efficient siRNA/BTZ co-delivery by the nanostructures inhibited cell viability and the clonogenic growth as well as stimulated apoptosis of AML cells. We hypothesized that SHARP1 downregulation induced the accumulation of non-functional MLL-AF6, DOT1L, MEN1, and LEDGF fusion proteins, preventing MLL-AF complex formation and downregulating RAS-GTP and Bcl-2 expression, consequently triggering autophagy and apoptosis. The BTZ combination substantially augmented therapeutic synergy and enhanced autophagic and apoptotic events. Our findings demonstrate a state-of-the-art biodegradable nanoplatform for siRNA/BTZ co-delivery with targeted SHARP1 knockdown, demonstrating a potential therapeutic option for MLL-AF6 AML.
    Keywords:  Acute myeloid leukemia; Apoptosis; Nanostructures; SHARP1; siRNA/bortezomib co-delivery
    DOI:  https://doi.org/10.1016/j.msec.2022.112663
  7. Mol Pharm. 2022 May 18.
    Exciting Times for Lipid Nanoparticles: How Canadian Discoveries Are Enabling Gene Therapies.
    Miffy H Y Cheng, Cedric A Brimacombe, Rein Verbeke, Pieter R Cullis.
      In this brief perspective, we describe key events in the history of the lipid-based nanomedicine field, highlight Canadian contributions, and outline areas where lipid nanoparticle technology is poised to have a transformative effect on the future of medicine.
    Keywords:  CAR-T therapy; CRISPR-Cas9; gene therapy; lipid nanoparticles; mRNA; vaccines
    DOI:  https://doi.org/10.1021/acs.molpharmaceut.2c00365
  8. Eur J Pharm Biopharm. 2022 May 15. pii: S0939-6411(22)00092-3. [Epub ahead of print]
    Structure-based peptide ligand design for improved epidermal growth factor receptor targeted gene delivery.
    Simon Decker, Alexander Taschauer, Emanuela Geppl, Viktoria Pirhofer, Michael Schauer, Stephan Pöschl, Florian Kopp, Lars Richter, Gerhard F Ecker, Haider Sami, Manfred Ogris.
      The epidermal growth factor receptor EGFR allows targeted delivery of macromolecular drugs to tumors. Its ligand, epidermal growth factor, binds EGFR with high affinity but acts mitogenic. Non-mitogenic peptides are utilized as targeting ligands, like the dodecapeptide GE11, although its low binding affinity warrants improvement. We applied a two-step computational approach with database search and molecular docking to design GE11 variants with improved binding. Synthesized peptides underwent binding studies on immobilized EGFR using surface plasmon resonance. Conjugates of peptides coupled via heterobifunctional PEG linker to linear polyethylenimine (LPEI) were used for transfection studies on EGFR-overexpressing cells using reporter gene encoding plasmid DNA. Docking studies unraveled similarities between GE11 and the EGFR dimerization arm. By skipping non-overlapping amino acids, a less hydrophobic segment (YTPQNVI) was identified being directly involved in EGFR binding. By replacing valine by tyrosine, a full-length version with proposed enhanced binding (GE11m3) was developed. While hydrophobic or hydrophilic segments and variations thereof exhibited low binding, GE11m3 exhibited 3-fold increase in binding compared to GE11, validating in silico predictions. In transfection studies, polyplexes with GE11m3 induced a significantly higher reporter gene expression when compared to GE11 polyplexes both on murine and human cancer cells overexpressing EGFR.
    Keywords:  EGFR; GE11; Polyplex; targeted gene delivery; transfection
    DOI:  https://doi.org/10.1016/j.ejpb.2022.05.004
  9. Front Immunol. 2022 ;13 856463
    PreS/2-21-Guided siRNA Nanoparticles Target to Inhibit Hepatitis B Virus Infection and Replication.
    Lixia Gao, Jie Yang, Jutao Feng, Ziying Liu, Ying Dong, Jiangyan Luo, Liangzhentian Yu, Jiamei Wang, Hongying Fan, Weifeng Ma, Tiancai Liu.
      A viable therapy is needed to overcome the deadlock of the incurable chronic hepatitis B (CHB). The prolonged existence of covalently closed circular DNA (cccDNA) and integrated HBV DNA in the nucleus of hepatocytes is the root cause of CHB. As a result, it is critical to successfully suppress HBV DNA replication and eliminate cccDNA. RNA interference has been proven in recent research to silence the expression of target genes and thereby decrease HBV replication. However, siRNA is susceptible to be degraded by RNA enzymes in vivo, making it difficult to deliver successfully and lacking of tissue targeting. To exploit the advantages of siRNA technology while also overcoming its limitations, we designed a new strategy and prepared biomimetic nanoparticles that were directed by PreS/2-21 peptides and precisely loaded HBV siRNA. Experiments on these nanoparticles in vitro and in vivo revealed that they are tiny, stable, safe and highly targetable, with high inhibitory effects on HBV DNA, pgRNA, cccDNA, HBeAg and HBsAg. PreS/2-21-directed nanoparticles loaded with HBV gene therapy drugs are expected to be promising for the treatment of CHB.
    Keywords:  HBV; liposomal nanoparticles; peptide; siRNA; targeting
    DOI:  https://doi.org/10.3389/fimmu.2022.856463
  10. Clin Cancer Res. 2022 May 18. pii: clincanres.3304.2022. [Epub ahead of print]
    RNA in cancer immunotherapy: unlocking the potential of the immune system.
    Wout de Mey, Arthur Esprit, Kris Thielemans, Karine Breckpot, Lorenzo Franceschini.
      Recent advances in the manufacturing, modification, purification and cellular delivery of ribonucleic acid (RNA) have enabled the development of RNA-based therapeutics for a broad array of applications. The approval of two SARS-CoV-2-targeting mRNA-based vaccines has highlighted the advances of this technology. Offering rapid and straightforward manufacturing, clinical safety and versatility, this paves the way for RNA therapeutics to expand into cancer immunotherapy. Together with ongoing trials on RNA cancer vaccination and cellular therapy, RNA therapeutics could be introduced into clinical practice, possibly stewarding future personalized approaches. In the present review, we discuss recent advances in RNA-based immuno-oncology together with an update on ongoing clinical applications and their current challenges.
    DOI:  https://doi.org/10.1158/1078-0432.CCR-21-3304
  11. Cancer Treat Rev. 2022 May 05. pii: S0305-7372(22)00069-X. [Epub ahead of print]107 102405
    The paradigm shift in treatment from Covid-19 to oncology with mRNA vaccines.
    Jiao Wei, Ai-Min Hui.
      mRNA vaccines have gained popularity over the last decade as a versatile tool for developing novel therapeutics. The recent success of coronavirus disease (COVID-19) mRNA vaccine has unlocked the potential of mRNA technology as a powerful therapeutic platform. In this review, we apprise the literature on the various types of cancer vaccines, the novel platforms available for delivery of the vaccines, the recent progress in the RNA-based therapies and the evolving role of mRNA vaccines for various cancer indications, along with a future strategy to treat the patients. Literature reveals that despite multifaceted challenges in the development of mRNA vaccines, the promising and durable efficacy of the RNA in pre-clinical and clinical studies deserves consideration. The introduction of mRNA-transfected DC vaccine is an approach that has gained interest for cancer vaccine development due to its ability to circumvent the necessity of DC isolation, ex vivo cultivation and re-infusion. The selection of appropriate antigen of interest remains one of the major challenges for cancer vaccine development. The rapid development and large-scale production of mRNA platform has enabled for the development of both personalized vaccines (mRNA 4157, mRNA 4650 and RO7198457) and tetravalent vaccines (BNT111 and mRNA-5671). In addition, mRNA vaccines combined with checkpoint modulators and other novel medications that reverse immunosuppression show promise, however further research is needed to discover which combinations are most successful and the best dosing schedule for each component. Each delivery route (intradermal, subcutaneous, intra tumoral, intranodal, intranasal, intravenous) has its own set of challenges to overcome, and these challenges will decide the best delivery method. In other words, while developing a vaccine design, the underlying motivation should be a reasonable combination of delivery route and format. Exploring various administration routes and delivery route systems has boosted the development of mRNA vaccines.
    Keywords:  Cancer vaccine; Covid-19; Oncology; Optimization; mRNA
    DOI:  https://doi.org/10.1016/j.ctrv.2022.102405
  12. Cancer Drug Resist. 2020 ;3(4): 930-946
    Albumin-based nanoparticles: a promising strategy to overcome cancer drug resistance.
    Islam Hassanin, Ahmed Elzoghby.
      Circumvention of cancer drug resistance is one of the major investigations in nanomedicine. In this regard, nanotechnology-based drug delivery has offered various implications. However, protein-based nanocarriers have been a versatile choice compared to other nanomaterials, provided by their favorable characteristics and safety profiles. Specifically, albumin-based nanoparticles have been demonstrated to be an effective drug delivery system, owing to the inherent targeting modalities of albumin, through gp60- and SPARC-mediated receptor endocytosis. Furthermore, surface functionalization was exploited for active targeting, due to albumin's abundance of carboxylic and amino groups. Stimuli-responsive drug release has also been pertained to albumin nano-systems. Therefore, albumin-based nanocarriers could potentially overcome cancer drug resistance through bypassing drug efflux, enhancing drug uptake, and improving tumor accumulation. Moreover, albumin nanocarriers improve the stability of various therapeutic cargos, for instance, nucleic acids, which allows their systemic administration. This review highlights the recent applications of albumin nanoparticles to overcome cancer drug resistance, the nano-fabrication techniques, as well as future perspectives and challenges.
    Keywords:  Albumin; active targeting; cancer therapy; drug delivery; multi-drug resistance; nanoparticles; nucleic acid therapy; stimuli- response release
    DOI:  https://doi.org/10.20517/cdr.2020.68
  13. Med (N Y). 2022 Mar 11. pii: S2666-6340(22)00084-8. [Epub ahead of print]3(3): 167-187
    Nanoscale delivery platforms for RNA therapeutics: Challenges and the current state of the art.
    Luke H Rhym, Daniel G Anderson.
      RNA nanomedicines present a promising class of therapeutics, with broad applications in protein replacement therapy, gene editing, immunotherapy, and vaccines, owing to their versatility and precise nature. Although recent years have seen dramatic improvements in the safety and efficacy of RNA-based therapies, their functional delivery to target tissues and cells in vivo remains challenging. Here, we discuss many of these challenges, as well as the methods and materials that have been developed to overcome them, with a focus on polymeric and lipid-based nanoscale delivery systems. In addition, we provide an overview of recent clinical and pre-clinical developments in RNA nanomedicines that have been made possible by advances in delivery.
    DOI:  https://doi.org/10.1016/j.medj.2022.02.001
This page is being maintained by Thomas Krichel. It was last updated on 2022‒05‒23 at 13:05:59.