bims-novged 2023-07-02 papers

bims-novged

Biomed News

on Non-viral vectors for gene delivery

Issue of 2023–07–02
twelve papers selected by
the Merkel lab, Ludwig-Maximilians University

Lipid nanoparticle topology regulates endosomal escape and delivery of RNA to the cytoplasm.
Membrane-destabilizing ionizable lipid empowered imaging-guided siRNA delivery and cancer treatment.
Screening Libraries to Discover Molecular Design Principles for the Targeted Delivery of mRNA with One-Component Ionizable Amphiphilic Janus Dendrimers Derived from Plant Phenolic Acids.
Polyplex Designs for Improving the Stability and Safety of RNA Therapeutics.
The Effect of Ethanol on Lipid Nanoparticle Stabilization from a Molecular Dynamics Simulation Perspective.
Novel Multi-Responsive Hyperbranched Polyelectrolyte Polyplexes as Potential Gene Delivery Vectors.
Anti-PEG IgM production induced by PEGylated liposomes as a function of administration route.
Inhalable siRNA Nanoparticles for Enhanced Tumor-Targeting Treatment of KRAS-Mutant Non-Small-Cell Lung Cancer.
Hyaluronic acid-functionalized lipoplexes and polyplexes as emerging nanocarriers for receptor-targeted cancer therapy.
Structural insights on ionizable Dlin-MC3-DMA lipids in DOPC layers by combining accurate atomistic force fields, molecular dynamics simulations and neutron reflectivity.
Smart Nanocarriers for the Targeted Delivery of Therapeutic Nucleic Acid for Cancer Immunotherapy.
Nanoparticle Brushes: Macromolecular Ligands for Materials Synthesis.

Proc Natl Acad Sci U S A. 2023 07 04. 120(27): e2301067120

Lipid nanoparticle topology regulates endosomal escape and delivery of RNA to the cytoplasm.

Lining Zheng, Sarith R Bandara, Zhengzhong Tan, Cecilia Leal.

  RNA therapeutics have the potential to resolve a myriad of genetic diseases. Lipid nanoparticles (LNPs) are among the most successful RNA delivery systems. Expanding their use for the treatment of more genetic diseases hinges on our ability to continuously evolve the design of LNPs with high potency, cellular-specific targeting, and low side effects. Overcoming the difficulty of releasing cargo from endocytosed LNPs remains a significant hurdle. Here, we investigate the fundamental properties of nonviral RNA nanoparticles pertaining to the activation of topological transformations of endosomal membranes and RNA translocation into the cytosol. We show that, beyond composition, LNP fusogenicity can be prescribed by designing LNP nanostructures that lower the energetic cost of fusion and fusion-pore formation with a target membrane. The inclusion of structurally active lipids leads to enhanced LNP endosomal fusion, fast evasion of endosomal entrapment, and efficacious RNA delivery. For example, conserving the lipid make-up, RNA-LNPs having cuboplex nanostructures are significantly more efficacious at endosomal escape than traditional lipoplex constructs.

Keywords:  RNA delivery; cubosomes; endosomal escape; lipids; membrane-fusion

DOI:  https://doi.org/10.1073/pnas.2301067120
Exploration (Beijing). 2021 Aug;1(1): 35-49

Membrane-destabilizing ionizable lipid empowered imaging-guided siRNA delivery and cancer treatment.

Shuai Guo, Kun Li, Bo Hu, Chunhui Li, Mengjie Zhang, Abid Hussain, Xiaoxia Wang, Qiang Cheng, Feng Yang, Kun Ge, Jinchao Zhang, Jin Chang, Xing-Jie Liang, Yuhua Weng, Yuanyu Huang.

  One of the imperative medical requirements for cancer treatment is how to establish an imaging-guided nanocarrier that combines therapeutic and imaging agents into one system. siRNA therapeutics have shown promising prospects in controlling life-threatening diseases. However, it is still challenging to develop siRNA formulations with excellent cellular entry capability, efficient endosomal escape, and simultaneous visualization. Herein, we fabricated multifunctional ionizable lipid nanoparticles (iLNPs) for targeted delivery of siRNA and MRI contrast agent. The iLNPs comprises DSPC, cholesterol, PEGylated lipid, contrast agent DTPA-BSA (Gd), and ionizable lipid termed iBL0104. siRNA-loaded iLNPs (iLNPs/siRNA) could be decorated with a tumor targeting cyclic peptide (c(GRGDSPKC)) (termed GARP), or without targeting modification (termed GAP). Data revealed that GARP/siRNA iLNPs exhibited significantly higher cellular entry efficiency than GAP/siRNA iLNPs. GARP/siRNA iLNPs rapidly and effectively escaped from endosome and lysosome after internalization. Compared with GAP/siPLK1, GARP/siPLK1 exhibited better tumor inhibition efficacy in both cell-line derived xenograft and liver cancer patient derived xenograft murine models. In addition, GARP formulation displayed ideal MRI effect in tumor-bearing mice, and was well tolerated by testing animals. Therefore, this study provides an excellent example for achieving imaging-guided and tumor-targeted siRNA delivery and cancer treatment, highlighting its promising potential for translational medicine application.

Keywords:  MRI; cancer treatment; hepatocellular carcinoma; lipid nanoparticle; lipid‐like material; siRNA

DOI:  https://doi.org/10.1002/EXP.20210008
Pharmaceutics. 2023 May 23. pii: 1572. [Epub ahead of print]15(6):

Screening Libraries to Discover Molecular Design Principles for the Targeted Delivery of mRNA with One-Component Ionizable Amphiphilic Janus Dendrimers Derived from Plant Phenolic Acids.

Juncheng Lu, Elena N Atochina-Vasserman, Devendra S Maurya, Muhammad Irhash Shalihin, Dapeng Zhang, Srijay S Chenna, Jasper Adamson, Matthew Liu, Habib Ur Rehman Shah, Honey Shah, Qi Xiao, Bryn Queeley, Nathan A Ona, Erin K Reagan, Houping Ni, Dipankar Sahoo, Mihai Peterca, Drew Weissman, Virgil Percec.

  Viral and synthetic vectors to deliver nucleic acids were key to the rapid development of extraordinarily efficient COVID-19 vaccines. The four-component lipid nanoparticles (LNPs), containing phospholipids, PEG-conjugated lipids, cholesterol, and ionizable lipids, co-assembled with mRNA via a microfluidic technology, are the leading nonviral delivery vector used by BioNTech/Pfizer and Moderna to access COVID-19 mRNA vaccines. LNPs exhibit a statistical distribution of their four components when delivering mRNA. Here, we report a methodology that involves screening libraries to discover the molecular design principles required to realize organ-targeted mRNA delivery and mediate activity with a one-component ionizable multifunctional amphiphilic Janus dendrimer (IAJD) derived from plant phenolic acids. IAJDs co-assemble with mRNA into monodisperse dendrimersome nanoparticles (DNPs) with predictable dimensions, via the simple injection of their ethanol solution in a buffer. The precise location of the functional groups in one-component IAJDs demonstrated that the targeted organs, including the liver, spleen, lymph nodes, and lung, are selected based on the hydrophilic region, while activity is associated with the hydrophobic domain of IAJDs. These principles, and a mechanistic hypothesis to explain activity, simplify the synthesis of IAJDs, the assembly of DNPs, handling, and storage of vaccines, and reduce price, despite employing renewable plant starting materials. Using simple molecular design principles will lead to increased accessibility to a large diversity of mRNA-based vaccines and nanotherapeutics.

Keywords:  dendrimersome nanoparticles; ionizable Janus dendrimer; molecular design principles; nanotherapeutics; one-component vector; plant phenolic acids; reduced vaccine price; simplified preparation and handling; targeted mRNA delivery; vaccines

DOI:  https://doi.org/10.3390/pharmaceutics15061572
Adv Drug Deliv Rev. 2023 Jun 24. pii: S0169-409X(23)00287-9. [Epub ahead of print] 114972

Polyplex Designs for Improving the Stability and Safety of RNA Therapeutics.

Satoshi Uchida, Chun Yin Jerry Lau, Makoto Oba, Kanjiro Miyata.

  Nanoparticle-based delivery systems have contributed to the recent clinical success of RNA therapeutics, including siRNA and mRNA. RNA delivery using polymers has several distinct properties, such as enabling RNA delivery into extra-hepatic organs, modulation of immune responses to RNA, and regulation of intracellular RNA release. However, delivery systems should overcome safety and stability issues to achieve widespread therapeutic applications. Safety concerns include direct damage to cellular components, innate and adaptive immune responses, complement activation, and interaction with surrounding molecules and cells in the blood circulation. The stability of the delivery systems should balance extracellular RNA protection and controlled intracellular RNA release, which requires optimization for each RNA species. Further, polymer designs for improving safety and stability often conflict with each other. This review covers advances in polymer-based approaches to address these issues over several years, focusing on biological understanding and design concepts for delivery systems rather than material chemistry.

Keywords:  Cytotoxicity; Innate immune response; Nanoparticle; Polycation; Polyion exchange reaction; RNA engineering; Ribonuclease; mRNA; siRNA

DOI:  https://doi.org/10.1016/j.addr.2023.114972
Molecules. 2023 Jun 17. pii: 4836. [Epub ahead of print]28(12):

The Effect of Ethanol on Lipid Nanoparticle Stabilization from a Molecular Dynamics Simulation Perspective.

Ari Hardianto, Zahra Silmi Muscifa, Wahyu Widayat, Muhammad Yusuf, Toto Subroto.

  Lipid nanoparticles (LNPs) have emerged as a promising delivery system, particularly for genetic therapies and vaccines. LNP formation requires a specific mixture of nucleic acid in a buffered solution and lipid components in ethanol. Ethanol acts as a lipid solvent, aiding the formation of the nanoparticle's core, but its presence can also affect LNP stability. In this study, we used molecular dynamics (MD) simulations to investigate the physicochemical effect of ethanol on LNPs and gain a dynamic understanding of its impact on the overall structure and stability of LNPs. Our results demonstrate that ethanol destabilizes LNP structure over time, indicated by increased root mean square deviation (RMSD) values. Changes in the solvent-accessible surface area (SASA), electron density, and radial distribution function (RDF) also suggest that ethanol affects LNP stability. Furthermore, our H-bond profile analysis shows that ethanol penetrates the LNP earlier than water. These findings emphasize the importance of immediate ethanol removal in lipid-based systems during LNP production to ensure stability.

Keywords:  ethanol; lipid nanoparticle; molecular dynamics simulation; stability

DOI:  https://doi.org/10.3390/molecules28124836
Pharmaceutics. 2023 May 30. pii: 1627. [Epub ahead of print]15(6):

Novel Multi-Responsive Hyperbranched Polyelectrolyte Polyplexes as Potential Gene Delivery Vectors.

Dimitrios Selianitis, Hector Katifelis, Maria Gazouli, Stergios Pispas.

  In this work, we investigate the complexation behavior of poly(oligo(ethylene glycol)methyl methacrylate)-co-poly(2-(diisopropylamino)ethyl methacrylate), P(OEGMA-co-DIPAEMA), hyperbranched polyelectrolyte copolymers, synthesized by reversible addition fragmentation chain transfer (RAFT) polymerization, with short-linear DNA molecules. The synthesized hyperbranched copolymers (HBC), having a different chemical composition, are prepared in order to study their ability to bind with a linear nucleic acid at various N/P ratios (amine over phosphate groups). Specifically, the three pH and thermo-responsive P(OEGMA-co-DIPAEMA) hyperbranched copolymers were able to form polyplexes with DNA, with dimensions in the nanoscale. Using several physicochemical methods, such as dynamic and electrophoretic light scattering (DLS, ELS), as well as fluorescence spectroscopy (FS), the complexation process and the properties of formed polyplexes were explored in response to physical and chemical stimuli such as temperature, pH, and ionic strength. The mass and the size of polyplexes are shown to be affected by the hydrophobicity of the copolymer utilized each time, as well as the N/P ratio. Additionally, the stability of polyplexes in the presence of serum proteins is found to be excellent. Finally, the multi-responsive hyperbranched copolymers were evaluated regarding their cytotoxicity via in vitro experiments on HEK 293 non-cancerous cell lines and found to be sufficiently non-toxic. Based on our results, these polyplexes could be useful candidates for gene delivery and related biomedical applications.

Keywords:  MTT assay; gene delivery; hyperbranched polyelectrolyte copolymers; pH/thermo-sensitive; polyplexes; short DNA

DOI:  https://doi.org/10.3390/pharmaceutics15061627
J Control Release. 2023 Jun 22. pii: S0168-3659(23)00400-5. [Epub ahead of print]

Anti-PEG IgM production induced by PEGylated liposomes as a function of administration route.

Haruka Takata, Taro Shimizu, Rina Yamade, Nehal E Elsadek, Sherif E Emam, Hidenori Ando, Yu Ishima, Tatsuhiro Ishida.

  Modifying the surface of nanoparticles with polyethylene glycol (PEG) is a commonly used approach for improving the in vitro stability of nanoparticles such as liposomes and increasing their circulation half-lives. We have demonstrated that, in certain conditions, an intravenous (i.v.) injection of PEGylated liposomes (PEG-Lip) induced anti-PEG IgM antibodies, which led to rapid clearance of second doses in mice. SARS-CoV-2 vaccines, composed of mRNA-containing PEGylated lipid nanoparticles, have been widely administered as intramuscular (i.m.) injections, so it is important to determine if PEGylated formulations can induce anti-PEG antibodies. If the favorable properties that PEGylation imparts to therapeutic nanoparticles are to be widely applicable this should apply to various routes of administration. However, there are few reports on the effect of different administration routes on the in vivo production of anti-PEG IgM. In this study, we investigated anti-PEG IgM production in mice following i.m., intraperitoneal (i.p.) and subcutaneous (s.c.) administration of PEG-Lip. PEG-Lip appeared to induce anti-PEG IgM by all the tested routes of administration, although the lipid dose causing maximum responses varied. Splenectomy attenuated the anti-PEG IgM production for all routes of administration, suggesting that splenic immune cells may have contributed to anti-PEG IgM production. Interestingly, in vitro experiments indicated that not only splenic cells but also cells in the peritoneal cavity induced anti-PEG IgM following incubation with PEG-Lip. These observations confirm previous experiments that have shown that measurable amounts of PEG-Lip administered i.p., i.m. or s.c. are absorbed to some extent into the blood circulation, where they can be distributed to the spleen and/or peritoneal cavity, and are recognized by B cells, triggering anti-PEG IgM production. The results obtained in this study have important implications for developing efficient PEGylated nanoparticular delivery system.

Keywords:  Administration route; Anti-PEG IgM; PEGylated nanoparticles; Polyethylene glycol (PEG); T cell independent (TI) antigen

DOI:  https://doi.org/10.1016/j.jconrel.2023.06.027
ACS Appl Mater Interfaces. 2023 Jun 24.

Inhalable siRNA Nanoparticles for Enhanced Tumor-Targeting Treatment of KRAS-Mutant Non-Small-Cell Lung Cancer.

Guolin Zhao, William Ho, Jinxian Chu, Xiaojian Xiong, Bin Hu, Kofi Oti Boakye-Yiadom, Xiaoyang Xu, Xue-Qing Zhang.

  Kirsten rat sarcoma (KRAS) is the most commonly mutated oncogene in lung cancers. Gene therapy is emerging as a promising cancer treatment modality; however, the systemic administration of gene therapy has been limited by inefficient delivery to the lungs and systemic toxicity. Herein, we report a noninvasive aerosol inhalation nanoparticle (NP) system, termed "siKRAS@GCLPP NPs," to treat KRAS-mutant non-small-cell lung cancer (NSCLC). The self-assembled siKRAS@GCLPP NPs are capable of maintaining structural integrity during nebulization, with preferential distribution within the tumor-bearing lung. Inhalable siKRAS@GCLPP NPs show not only significant tumor-targeting capability but also enhanced antitumor activity in an orthotopic mouse model of human KRAS-mutant NSCLC. The nebulized delivery of siKRAS@GCLPP NPs demonstrates potent knockdown of mutated KRAS in tumor-bearing lungs without causing any observable adverse effects, exhibiting a better biosafety profile than the systemic delivery approach. The results present a promising inhaled gene therapy approach for the treatment of KRAS-mutant NSCLC and other respiratory diseases.

Keywords:  KRAS mutation; inhaled siRNA therapeutics; lung cancer therapy; pulmonary nucleic acid delivery; tumor-targeting gene therapy

DOI:  https://doi.org/10.1021/acsami.3c05007
Environ Res. 2023 Jun 25. pii: S0013-9351(23)01310-5. [Epub ahead of print]233 116506

Hyaluronic acid-functionalized lipoplexes and polyplexes as emerging nanocarriers for receptor-targeted cancer therapy.

Jyoti Chandra, Nagashekhara Molugulu, Sivakumar Annadurai, Shadma Wahab, Ritu Karwasra, Surender Singh, Rahul Shukla, Prashant Kesharwani.

  Cancer is an intricate disease that develops as a response to a combination of hereditary and environmental risk factors, which then result in a variety of changes to the genome. The cluster of differentiation (CD44) is a type of transmembrane glycoprotein that serves as a potential biomarker for cancer stem cells (CSC) and viable targets for therapeutic intervention in the context of cancer therapy. Hyaluronic acid (HA) is a linear polysaccharide that exhibits a notable affinity for the CD44 receptor. This characteristic renders it a promising candidate for therapeutic interventions aimed at selectively targeting CD44-positive cancer cells. Treating cancer via non-viral vector-based gene delivery has changed the notion of curing illness through the incorporation of therapeutic genes into the organism. The objective of this review is to provide an overview of various hyaluronic acid-modified lipoplexes and polyplexes as potential drug delivery methods for specific forms of cancer by effectively targeting CD44.

Keywords:  CD44; Cancer stem cell; Gene delivery; Lipoplexes; Polyplexes

DOI:  https://doi.org/10.1016/j.envres.2023.116506
Nanoscale. 2023 Jun 28.

Structural insights on ionizable Dlin-MC3-DMA lipids in DOPC layers by combining accurate atomistic force fields, molecular dynamics simulations and neutron reflectivity.

Mohd Ibrahim, Jennifer Gilbert, Marcel Heinz, Tommy Nylander, Nadine Schwierz.

Ionizable lipids such as the promising Dlin-MC3-DMA (MC3) are essential for the successful design of lipid nanoparticles (LNPs) as drug delivery agents. Combining molecular dynamics simulations with experimental data, such as neutron reflectivity experiments and other scattering techniques, is essential to provide insights into the internal structure of LNPs, which is not fully understood to date. However, the accuracy of the simulations relies on the choice of force field parameters and high-quality experimental data is indispensable to verify the parametrization. For MC3, different parameterizations in combination with the CHARMM and the Slipids force fields have recently emerged. Here, we complement the existing efforts by providing parameters for cationic and neutral MC3 compatible with the AMBER Lipid17 force field. Subsequently, we carefully assess the accuracy of the different force fields by providing a direct comparison to neutron reflectivity experiments of mixed lipid bilayers consisting of MC3 and DOPC at different pHs. At low pH (cationic MC3) and at high pH (neutral MC3) the newly developed MC3 parameters in combination with AMBER Lipid17 for DOPC give good agreement with the experiments. Overall, the agreement is similar compared to the Park-Im parameters for MC3 in combination with the CHARMM36 force field for DOPC. The Ermilova-Swenson MC3 parameters in combination with the Slipids force field underestimate the bilayer thickness. While the distribution of cationic MC3 is very similar, the different force fields for neutral MC3 reveal distinct differences ranging from strong accumulation in the membrane center (current MC3/AMBER Lipid17 DOPC), over mild accumulation (Park-Im MC3/CHARMM36 DOPC) to surface accumulation (Ermilova-Swenson MC3/Slipids DOPC). These pronounced differences highlight the importance of accurate force field parameters and their experimental validation.

DOI: https://doi.org/10.1039/d3nr00987d
Pharmaceutics. 2023 Jun 15. pii: 1743. [Epub ahead of print]15(6):

Smart Nanocarriers for the Targeted Delivery of Therapeutic Nucleic Acid for Cancer Immunotherapy.

Abu Baker, Jochen Lorch, David VanderWeele, Bin Zhang.

  A wide variety of therapeutic approaches and technologies for delivering therapeutic agents have been investigated for treating cancer. Recently, immunotherapy has achieved success in cancer treatment. Successful clinical results of immunotherapeutic approaches for cancer treatment were led by antibodies targeting immune checkpoints, and many have advanced through clinical trials and obtained FDA approval. A major opportunity remains for the development of nucleic acid technology for cancer immunotherapy in the form of cancer vaccines, adoptive T-cell therapies, and gene regulation. However, these therapeutic approaches face many challenges related to their delivery to target cells, including their in vivo decay, the limited uptake by target cells, the requirements for nuclear penetration (in some cases), and the damage caused to healthy cells. These barriers can be avoided and resolved by utilizing advanced smart nanocarriers (e.g., lipids, polymers, spherical nucleic acids, metallic nanoparticles) that enable the efficient and selective delivery of nucleic acids to the target cells and/or tissues. Here, we review studies that have developed nanoparticle-mediated cancer immunotherapy as a technology for cancer patients. Moreover, we also investigate the crosstalk between the function of nucleic acid therapeutics in cancer immunotherapy, and we discuss how nanoparticles can be functionalized and designed to target the delivery and thus improve the efficacy, toxicity, and stability of these therapeutics.

Keywords:  cancer; immunotherapy; nanoparticles; nucleic acids; targeted delivery

DOI:  https://doi.org/10.3390/pharmaceutics15061743
Acc Chem Res. 2023 Jun 30.

Nanoparticle Brushes: Macromolecular Ligands for Materials Synthesis.

Theodore Hueckel, Xin Luo, Omar F Aly, Robert J Macfarlane.

ConspectusColloidal nanoparticles have unique attributes that can be used to synthesize materials with exotic properties, but leveraging these properties requires fine control over the particles' interactions with one another and their surrounding environment. Small molecules adsorbed on a nanoparticle's surface have traditionally served as ligands to govern these interactions, providing a means of ensuring colloidal stability and dictating the particles' assembly behavior. Alternatively, nanoscience is increasingly interested in instead using macromolecular ligands that form well-defined polymer brushes, as these brushes provide a much more tailorable surface ligand with significantly greater versatility in both composition and ligand size. While initial research in this area is promising, synthesizing macromolecules that can appropriately form brush architectures remains a barrier to their more widespread use and limits understanding of the fundamental chemical and physical principles that influence brush-grafted particles' ability to form functional materials. Therefore, enhancing the capabilities of polymer-grafted nanoparticles as tools for materials synthesis requires a multidisciplinary effort, with specific focus on both developing new synthetic routes to polymer-brush-coated nanoparticles and investigating the structure-property relationships the brush enables.In this Account, we describe our recent work in developing polymer brush coatings for nanoparticles, which we use to modulate particle behavior on demand, select specific nanoscopic architectures to form, and bolster traditional bulk polymers to form stronger materials by design. Distinguished by the polymer type and capabilities, three classes of nanoparticles are discussed here: nanocomposite tectons (NCTs), which use synthetic polymers end-functionalized with supramolecular recognition groups capable of directing their assembly; programmable atom equivalents (PAEs) containing brushes of synthetic DNA that employ Watson-Crick base pairing to encode particle binding interactions; and cross-linkable nanoparticles (XNPs) that can both stabilize nanoparticles in solution and polymer matrices and subsequently form multivalent cross-links to strengthen polymer composites. We describe the formation of these brushes through "grafting-from" and "grafting-to" strategies and illustrate aspects that are important for future advancement. We also examine the new capabilities brushes provide, looking closely at dynamic polymer processes that provide control over the assembly state of particles. Finally, we provide a brief overview of the technological applications of nanoparticles with polymer brushes, focusing on the integration of nanoparticles into traditional materials and the processing of nanoparticles into bulk solids.

DOI: https://doi.org/10.1021/acs.accounts.3c00160