Lipid Nanoparticle-Based Inhibitors for SARS-CoV-2 Host Cell Infection
Vinith Yathindranath, Nura Safa, Mateusz Marek Tomczyk, Vernon Dolinsky, Donald W Miller
International Journal of Nanomedicine, doi:10.2147/ijn.s448005
Purpose: The global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the lingering threat to public health has fueled the search for effective therapeutics to treat SARS-CoV-2. This study aimed to develop lipid nanoparticle (LNP) inhibitors of SARS-CoV-2 entry to reduce viral infection in the nose and upper airway. Methods: Two types of LNP formulations were prepared following a microfluidic mixing method. The LNP-Trap consisted of DOPC, DSPC, cholesterol, and DSPE-PEG-COOH modified with various spike protein binding ligands, including ACE2 peptide, recombinant human ACE2 (rhACE2) or monoclonal antibody to spike protein (mAb). The LNP-Trim consisted of ionizing cationic DLin-MC3-DMA, DSPC, cholesterol, and DMG-PEG lipids encapsulating siACE2 or siTMPRSS2. Both formulations were assayed for biocompatibility and cell uptake in airway epithelial cells (Calu-3). Functional assessment of activity was performed using SARS-CoV-2 spike protein binding assays (LNP-Trap), host receptor knockdown (LNP-Trim), and SARS-CoV-2 pseudovirus neutralization assay (LNP-Trap and LNP-Trim). Localization and tissue distribution of fluorescently labeled LNP formulations were assessed in mice following intranasal administration. Results: Both LNP formulations were biocompatible based on cell impedance and MTT cytotoxicity studies in Calu-3 cells at concentrations as high as 1 mg/mL. LNP-Trap formulations were able to bind spike protein and inhibit pseudovirus infection by 90% in Calu-3 cells. LNP-Trim formulations reduced ACE2 and TMPRSS2 at the mRNA (70% reduction) and protein level (50% reduction). The suppression of host targets in Calu-3 cells treated with LNP-Trim resulted in over 90% inhibition of pseudovirus infection. In vivo studies demonstrated substantial retention of LNP-Trap and LNP-Trim in the nasal cavity following nasal administration with minimal systemic exposure. Conclusion: Both LNP-Trap and LNP-Trim formulations were able to safely and effectively inhibit SARS-CoV-2 pseudoviral infection in airway epithelial cells. These studies provide proof-of-principle for a localized treatment approach for SARS-CoV-2 in the upper airway.
Disclosure
References
Ahn, Kim, Hong, Nasal ciliated cells are primary targets for SARS-CoV-2 replication in the early stage of COVID-19, J Clin Investig,
doi:10.1172/JCI148517
Arias, Oliveros, Lechtig, Bustos, Biologics in COVID-19 so far: systematic review, Pharmaceuticals,
doi:10.3390/ph15070783
Baram-Pinto, Shukla, Gedanken, Sarid, Inhibition of HSV-1 attachment, entry, and cell-to-cell spread by functionalized multivalent gold nanoparticles, Small,
doi:10.1002/smll.200902384
Bayón-Cordero, Alkorta, Arana, Application of solid lipid nanoparticles to improve the efficiency of anticancer drugs, Nanomaterials,
doi:10.3390/nano9030474
Belliveau, Huft, Lin, Microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of siRNA, Mol Ther Nucleic Acids,
doi:10.1038/mtna.2012.28
Bowman, Ballard, Ackerson, Feldheim, Margolis et al., Inhibition of HIV fusion with multivalent gold nanoparticles, J Am Chem Soc,
doi:10.1021/ja710321g
Chan, Tan, Narayanan, Procko, An engineered decoy receptor for SARS-CoV-2 broadly binds protein S sequence variants, Sci Adv,
doi:10.1126/sciadv.abf1738
Chen, Cheng, Roffler, Polyethylene glycol immunogenicity: theoretical, clinical, and practical aspects of anti-polyethylene glycol antibodies, ACS Nano,
doi:10.1021/acsnano.1c05922
Chen, Fang, Chen, Targeting and enrichment of viral pathogen by cell membrane cloaked magnetic nanoparticles for enhanced detection, ACS Appl Mater Interfaces,
doi:10.1021/acsami.7b09931
El-Shennawy, Hoffmann, Dashzeveg, Circulating ACE2-expressing extracellular vesicles block broad strains of SARS-CoV-2, Nat Commun,
doi:10.1038/s41467-021-27893-2
Giovane, Rezai, Henderson, Current pharmacological modalities for management of novel coronavirus disease 2019 (COVID-19) and the rationale for their utilization: a review, Rev Med Virol,
doi:10.1002/rmv.2136
Gong, -J Y, PTD4-apoptin protein and dacarbazine show a synergistic antitumor effect on B16-F1 melanoma in vitro and in vivo, Eur J Pharmacol,
doi:10.1016/j.ejphar.2010.12.004
Gottlieb, Nirula, Chen, Effect of bamlanivimab as monotherapy or in combination with etesevimab on viral load in patients with mild to moderate COVID-19: a randomized clinical trial, JAMA,
doi:10.1001/jama.2021.0202
Hak, Helgesen, Hektoen, The effect of nanoparticle polyethylene glycol surface density on ligand-directed tumor targeting studied in vivo by dual modality imaging, ACS Nano,
doi:10.1021/nn301630n
Han, Penn-Nicholson, Cho, Identification of critical determinants on ACE2 for SARS-CoV entry and development of a potent entry inhibitor, International Journal of Nanomedicine,
doi:10.1016/j.virol.2006.01.029
Haschke, Schuster, Poglitsch, Pharmacokinetics and pharmacodynamics of recombinant human angiotensin-converting enzyme 2 in healthy human subjects, Clin Pharmacokinet,
doi:10.1007/s40262-013-0072-7
Hoffmann, Hofmann-Winkler, Smith, Camostat mesylate inhibits SARS-CoV-2 activation by TMPRSS2-related proteases and its metabolite GBPA exerts antiviral activity, EBioMedicine,
doi:10.1016/j.ebiom.2021.103255
Hoffmann, Kleine-Weber, Schroeder, SARS-CoV-2 cell entry depends on ACE2 and TMPRSS2 and is blocked by a clinically proven protease inhibitor, Cell,
doi:10.1016/j.cell.2020.02.052
Huang, Leobandung, Foss, Peppas, Molecular aspects of muco-and bioadhesion: tethered structures and site-specific surfaces, J Control Release,
doi:10.1016/S0168-3659(99)00233-3
Kuba, Yamaguchi, Penninger, Angiotensin-Converting Enzyme 2 (ACE2) in the pathogenesis of ARDS in COVID-19, Front Immunol,
doi:10.3389/fimmu.2021.732690
Martinez-Avila, Hijazi, Marradi, Gold manno-glyconanoparticies: multivalent systems to block HIV-1 gp120 binding to the lectin DC-SIGN, Chem Eur J,
doi:10.1002/chem.200900923
Mesias, Zhu, Tang, Effective ACE2 peptide-nanoparticle conjugation and its binding with the SARS-Cov-2 RBD quantified by dynamic light scattering, Chem Comm,
doi:10.1039/D1CC02267A
Nathan, Shawa, De, Torre, A narrative review of the clinical practicalities of bamlanivimab and etesevimab antibody therapies for SARS-CoV-2, Infect Dis Ther,
doi:10.1007/s40121-021-00515-6
Nikiforuk, Kuchinski, Twa, The contrasting role of nasopharyngeal angiotensin converting enzyme 2 (ACE2) transcription in SARS-CoV-2 infection: a cross-sectional study of people tested for COVID-19 in British Columbia, Canada, International Journal of Nanomedicine,
doi:10.1016/j.ebiom.2021.103316
Papp, Sieben, Ludwig, Inhibition of influenza virus infection by multivalent sialic-acid-functionalized gold nanoparticles, Small,
doi:10.1002/smll.201001349
Politch, Cu-Uvin, Moench, Safety, acceptability, and pharmacokinetics of a monoclonal antibody-based vaginal multipurpose prevention film (MB66): a Phase I randomized trial, PLoS Med,
doi:10.1371/journal.pmed.1003495
Pustake, Tambolkar, Giri, Gandhi, SARS, MERS and CoVID-19: an overview and comparison of clinical, laboratory and radiological features, J Family Med Prim Care,
doi:10.4103/jfmpc.jfmpc_839_21
Rao, Xia, Xu, Decoy nanoparticles protect against COVID-19 by concurrently adsorbing viruses and inflammatory cytokines, Proc Natl Acad Sci,
doi:10.1073/pnas.2014352117
Ren, Wang, Gao, Zhou, Omicron variant (B.1.1.529) of SARS-CoV-2: mutation, infectivity, transmission, and vaccine resistance, World J Clin Cases,
doi:10.12998/wjcc.v10.i1.1
Schütz, Ruiz-Blanco, Münch, Kirchhoff, Sanchez-Garcia et al., Peptide and peptide-based inhibitors of SARS-CoV-2 entry, Adv Drug Deliv Rev,
doi:10.1016/j.addr.2020.11.007
Semple, Klimuk, Harasym, Efficient encapsulation of antisense oligonucleotides in lipid vesicles using ionizable aminolipids: formation of novel small multilamellar vesicle structures, Biochim Biophys Acta,
doi:10.1016/S0005-2736(00)00343-6
Serra, Doménech, Peppas, Design of poly(ethylene glycol)-tethered copolymers as novel mucoadhesive drug delivery systems, CAS, SciSearch ® , Current Contents ® /Clinical Medicine, Journal Citation Reports/Science Edition,
doi:10.1016/j.ejpb.2005.10.011
Sullivan, Gebo, Shoham, Early outpatient treatment for Covid-19 with convalescent plasma, N Engl J Med,
doi:10.1056/NEJMoa2119657
Sungnak, Huang, Bécavin, SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes, Nat Med,
doi:10.1038/s41591-020-0868-6
Tam, Kulkarni, An, Lipid nanoparticle formulations for optimal RNA-based topical delivery to murine airways, Eur J Pharm Sci,
doi:10.1016/j.ejps.2022.106234
V'kovski, Kratzel, Steiner, Stalder, Thiel, Coronavirus biology and replication: implications for SARS-CoV-2, Nat Rev Microbiol,
doi:10.1038/s41579-020-00468-6
Valenti, Antonini, Lactoferrin: an important host defence against microbial and viral attack. Cellular and molecular life sciences, Cell Mol Life Sci,
doi:10.1007/s00018-005-5372-0
Van Der Meel, Chen, Zaifman, Modular lipid nanoparticle platform technology for siRNA and lipophilic prodrug delivery, Small,
doi:10.1002/smll.202103025
Wotring, Fursmidt, Ward, Sexton, Evaluating the in vitro efficacy of bovine lactoferrin products against SARS-CoV-2 variants of concern, J Dairy Sci,
doi:10.3168/jds.2021-21247
Xu, Ensign, Boylan, Impact of Surface Polyethylene Glycol (PEG) density on biodegradable nanoparticle transport in mucus ex vivo and distribution in vivo, ACS Nano,
doi:10.1021/acsnano.5b03876
Yathindranath, Safa, Sajesh, Spermidine/Spermine N1-Acetyltransferase 1 (SAT1)-A potential gene target for selective sensitization of glioblastoma cells using an ionizable lipid nanoparticle to deliver siRNA, Cancers,
doi:10.3390/cancers14215179
Álvarez-Viñas, Souto, Flórez-Fernández, Torres, Bandín et al., Antiviral activity of carrageenans and processing implications, Mar Drugs,
doi:10.3390/md19080437