The therapeutic use of messenger RNA (mRNA) has fueled great hope to combat a wide range of incurable diseases. RNA Therapeutics comprise a rapidly expanding category of drugs that will speed solutions to the clinic; will actualize personalized medicine; and will make the term undruggable obsolete. They also demonstrated HIV gag specific CD8+ and CD4+ T cell responses after immunization with PSA/mRNA vaccine (Zhao et al., 2016). 76, 37913799. mRNA therapies: new hope in the fight against melanoma: published as part of the Biochemistry series Biochemistry to Bedside.. Ther. Available online at: https://clinicaltrials.gov/ct2/show/NCT03897881 (accessed July 10, 2020). Soc. Rev. For the Cre-Ai9 mouse model used in Fig. medRxiv [Preprint]. To achieve this, we analyze the role of different helper lipids and component ratios in plasmid DNA lipid nanoparticle-mediated gene delivery in vitro and in vivo. 13, 2003. Akinc, A. et al. Exploring the pH sensitivity of Poly(allylamine) phosphate supramolecular nanocarriers for intracellular siRNA delivery. doi: 10.1007/s10096-020-03899-4. 11, 3232 (2020). Although several dominating players in the RNA biopharma sector have emerged, new small biotech startups as well as academic groups with transformative ideas are propagating. Ishino et al. CTX001 is designed to treat both sickle cell disease and beta thalassemia. ISSN 2041-1723 (online). Because mRNA is inherently unstable it requires delivery vehicles that will protect the cargo from RNAase degradation. Rather, the intracellular trafficking steps including endolysosomal escape and pDNA release play a more critical role. Nanotechnol. Cancer Biol. To further understand the rate-limiting steps of the in vivo gene delivery process for pDNA LNPs, we examined the in vivo biodistribution profile, cellular uptake level, and lysosome escape capability for the top-performing formulations and compared them with formulations that were less effective but shared similar characteristics. doi: 10.7150/thno.25958, Kessler, J. (2016). 5f). 14, 839846. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. Mol. 383, 19201931. This page was last edited on 5 November 2022, at 23:36. CRISPRCas9 structures and mechanisms. FDA. U.S.A. 109, 1197511980. Nat. (2019). boats.com - new and used boats for sale #everythingboats. Cell Rep. 21, 35483558. doi: 10.4103/0250-474X.58169, Schmeer, M., Buchholz, T., and Schleef, M. (2017). 37, 11741185 (2019). Moreover, the significant difference observed between groups incubated with or without serum suggested that the discrepancies between in vivo and in vitro experiments may be closely related to serum-mediated opsonization of the LNPs. The investigators purified cytoplasmic mRNA from wild-type rat hypothalamus, or synthesized mRNA that encodes for vasopressin. (2017). Special report: the birth of biotechnology. [76], Despite the contrary perception of much of the public, psychedelic drugs are not addictive and are physiologically safe. A total of 30 fields were analyzed inside each well of the plates; and the well-averaged results were generated by averaging all the cells in all the fields in each well. To verify that the transfection efficiency of LNPs was not related to biodistribution, we checked the transfection efficiency of six LNPs by administering the same dose of the six LNPs via intrahepatic injection. (2016). Rapamycin-loaded biomimetic nanoparticles reverse vascular inflammation. This result showed that these clusters contained LNP formulations that could have potent liver-specific transfection. Biophys. Figure 5 demonstrates systemically delivered mRNA expressing hepatic protein (An et al., 2017). The dilution ratio for all antibodies listed above was 1:200 with staining buffer (ThermoFisher #00422226). Patient-customized oligonucleotide therapy for a rare genetic disease. Volanesorsen (Ionis Pharmaceuticals) was recently approved in Europe (Waylivra, EU conditional marketing authorization in May 2019). Enhancement of SMN2 Exon 7 inclusion by antisense oligonucleotides targeting the exon. These cells home to the patients bone marrow where CD34+ cells replicate and generate normal ADA protein to correct the deficiency (South et al., 2018). For example, 8L of a particle suspension at 25g pDNA/mL was pipetted into the 200L culture media in the wells. The drug targets aminolevulinate synthase 1 mRNA in liver, and reduces the levels of disease-causing neurotoxic intermediates aminolevulinic acid and porphobilinogen (Alnylam, 2020b; Givlaari, 2020). Increased dystrophin production with golodirsen in patients with Duchenne muscular dystrophy. Drug Discov. To quantify the mCherry+ or tdTom+ cells among different cell types in each organ, cell isolation and staining was performed, followed by flow cytometry analysis. Ortego, J., Generation of a Replication-Competent, Propagation-Defecient Virus Vector based on the Transmissible Gastroenteritis Coronavirus Genome, J. Each of the three major companies has dozens of new mRNA therapeutics under development, some of which have been mentioned in this review. Hemoglobinopathies. Alternatively, the AAV vector can be utilized to deliver interference RNA to downregulate the expression of a specific gene. However, the liposome delivery systems have flaws; (1) liposomes are less stable, and may fuse or leak RNA resulting in low efficiency of delivery; (2) they entrap less RNA; (3) they can be harmful if oxidized; and (4) are not consistent in size. Ther. The innate immune activation against pDNA has been reported to induce gene silencing and inflammation response. The LNP suspensions were concentrated to 200g/mL of pDNA by an Amicon Ultra-2 centrifugal filter unit with a MWCO of 100kDa. doi: 10.1016/j.vaccine.2019.04.074. Cancer Ther. doi: 10.1038/mt.2012.18, Bertoletti, A., and Tan, A. T. (2020). When characterizing luciferase as the reporter, cells were lysed by reporter lysis buffer (Promega) using two freeze-thaw cycles, with the lysate characterized by a luminometer upon addition of luciferin assay solution (Promega) against a standard curve generated using luciferase samples (Promega). (2017) explored the pH sensitivity of their nucleic acids carrier to trigger endosomal delivery and found that the carrier is stable at narrow pH range (79). doi: 10.1212/WNL.0000000000009233, Gilbert, W. (1986). Hotboxing involves multiple persons smoking cannabis in an enclosed space, such as a car, with no way for smoke to escape. All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. 99, 129137 (2016). Strimvelis uses the patients own CD34+ cells generated from hematopoietic stem cell (HSCs). B., Buus, S., Erbar, S., Wee, E. G., Sahin, U., et al. 17. Self-amplifying RNA vaccines give equivalent protection against influenza to mRNA vaccines but at much lower doses. (2020). 20, 4554. After incubation, any remaining lung tissue was homogenized. Infiltrating monocyte-derived macrophages and resident kupffer cells display different ontogeny and functions in acute liver injury. The CD34+ cells are then transduced with a gammaretrovirus vector carrying the gene for human adenosine deaminase (ADA), then reinfused into the patient. Biochemistry 59, 16501655. See Supplementary Tables16 for molar percentage used in the 32 formulations for each helper lipid. J. Pharm. 9, 3824238254. Four in ten likely voters are The same payload, luciferase (Luc) (50%) and mCherry (50%) pDNA, were encapsulated in these LNPs, which were administered intravenously at a total pDNA dose of 100g per mouse. 1 hospital in Texas. Luxturna is an AAV (serotype 2) therapy encoding the RPE65 gene which received FDA approval in December 2017 and European approval in 2018. Sci. The antibodies used include Brilliant Violet 605 anti-mouse CD45 (BioLegend #103140), PerCP-Cyanine 5.5 anti-mouse CD11b (BioLegend #101228), APC anti-mouse CD11c (BioLegend #117309), FITC anti-mouse CD3 (BioLegend #100203) and PE-Cyanine 7 anti-mouse CD19 (BioLegend #115519). (1998). Amgen (2020). 6b, c, e, f, h. Statistical P-values: No significance: NS; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. (2019). mRNA vaccines against H10N8 and H7N9 influenza viruses of pandemic potential are immunogenic and well tolerated in healthy adults in phase 1 randomized clinical trials. ADS Yescarta is another CD19-targeting CAR immunotherapy to treat large B cell lymphoma. By contrast, nucleic-acid based strategies avoid many of these limitations as they make use of the translational machinery of the mammalian cell. Systemic nanoparticle delivery of CRISPR-Cas9 ribonucleoproteins for effective tissue specific genome editing. 6, 893903 (1999). Liposomes are formed when materials containing polar head groups and non-polar tails (phospholipids) are dispersed in aqueous phase. Pipeline CureVac. Target Ther. Lipid nanoparticles hold great potential as an effective non-viral vector for nucleic acid-based gene therapy. Mice were provided with corncob as bedding. The efficacy of this drug is being evaluated in a phase II trial conducted by AstraZeneca. & Tzeng, S. Y. Poly(beta-amino ester)s as gene delivery vehicles: challenges and opportunities. The combination of lipid and polymer permits combination therapy. clinicaltrials.gov. J. Clin. 2-Fluoropyrimidine RNA-based Aptamers to the 165-Amino Acid Form of Vascular Endothelial Growth Factor (VEGF165) INHIBITION OF RECEPTOR BINDING AND VEGF-INDUCED VASCULAR PERMEABILITY THROUGH INTERACTIONS REQUIRING THE EXON 7-ENCODED DOMAIN. Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs. Gene Ther. Five of the 16 formulations (DI-2, DI-4, FIII-2, FIII-3 and FIII-4) showed high toxicity following i.v. doi: 10.1038/nrd1632. Using an optimized LNP formulation, we examined the effect of co-delivering pDNA and siRNAs against signal transducer and activator of transcription (STAT)26 and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-B)27,28,29 on the level and duration of transgene expression following i.v. The AAV vector delivers the desired gene to the target cell, for example to restore normal protein function (Tratschin et al., 1984; Naso et al., 2017). Similarly, Olaptesed pegol (NOXXON, 2020a; NOXXON Pharma) was evolved to target CXC chemokine ligand (CXCL12) for clinical treatment of brain cancer (glioblastoma/glioma). The PEG functionalization is crucial for this platform as it allows enough circulation time for the drug to be localized in the liver. a, b Whole-body bioluminescence imaging of female BALB/c mice (68 weeks) at different time points after i.v. These drugs are cost effective, 11:3523. doi: 10.1038/s41467-020-17409-9, McNamara, J. O., Andrechek, E. R., Wang, Y., Viles, K. D., Rempel, R. E., Gilboa, E., et al. Using this approach, Poseida Therapeutics has developed a cell therapy for multiple myeloma, P-BCMA-101, which is now in clinical trials. The Center for Biologics Evaluation and Research (CBER) is primarily responsible for evaluation and approval of vaccines, allergenic products, blood and blood products, plasma derivatives, cellular and gene therapy products. A 24h incubation was followed to allow transgene expression. 381, 16441652. The advantage of the self-replicating approach is that significantly higher amount of antigen can be expressed with lower doses of mRNA (Vogel et al., 2018). 50, 259293. Similarly, most clusters composed of anionic helper lipids had limited local transfection in the liver, but there was a unique cluster (FIII) in the 18PG group that achieved relatively high transfection efficiency. De Keersmaecker, B., Claerhout, S., Carrasco, J., Bar, I., Corthals, J., Wilgenhof, S., et al. Nat. In addition, higher levels of transgene expression in the spleen were observed for clusters CI (45.2%) and CII (30.0%). These nanoparticles degrade into non-toxic products and are safely excreted (Mller et al., 2016; Paris and Vallet-Reg, 2020). Int. ACS Comb. Clinical development of TargomiRs, a miRNA mimic-based treatment for patients with recurrent thoracic cancer. Safety and immunogenicity of two RNA-based Covid-19 vaccine candidates. Recent innovations in mRNA vaccines. Biodegradable polymeric nanoparticles based drug delivery systems. The Onpattro story and the clinical translation of nanomedicines containing nucleic acid-based drugs. 1, 147168. Fluorescent signal was also observed in other organs (Fig. In developing this multi-step composition screening process, we successfully identified the top performing LNP formulations from the designed library consisting of 1080 formulations for liver-targeted transfection through i.v. Pharmacol. doi: 10.1016/0378-1119(92)90121-5, Le, T. T., Andreadakis, Z., Kumar, A., Romn, R. G., Tollefsen, S., Saville, M., et al. (2005) observed that functionalized carbon nanotubes could deliver siRNA against lamin A/C to suppress the expression of this protein in HeLa cells. Furthermore, only 1014% of proteins have active binding sites that are druggable targets for small molecules (Hopkins and Groom, 2002). Schematic illustration of intravenous administration of mRNA encapsulated in lipid nanoparticles (LNP) to restore missing/defective protein in hepatic cells (An et al., 2017). Although there are no miRNA-based drugs on the market as of today, some promising candidates are currently in clinical trials. Multi-step screening of DNA/lipid nanoparticles and co-delivery with siRNA to enhance and prolong gene expression. Indian J. Pharm. Small 14:1702564. doi: 10.1002/smll.201702564, Lee, R. C., Feinbaum, R. L., and Ambros, V. (1993). CAS 5:e73. 1). 77, 13101321. Assembling OX40 aptamers on a molecular scaffold to create a receptor-activating aptamer. Kymriah is CD19-targeting CAR T cells immunotherapy to treat leukemia, lymphoma, and pediatric cancer in a single dose (approved in United States and Europe in 2017 and 2018 respectively). mRNA-based therapeuticsdeveloping a new class of drugs. Biotechnol., 18 March 2021, View all Recent advances with liposomes as pharmaceutical carriers. Nano Lett. The particles prepared were pipetted into EMEM medium at a final particle concentration of 1g pDNA/mL. RNA Therapeutics is a rapidly emerging field of biotherapeutics. MRT5005 encodes fully functional CFTR and is delivered to lung epithelial cells through nebulization (TranslateBio, 2019). Different methods to determine the encapsulation efficiency of protein in PLGA nanoparticles. FDA (2021a). Determination of the role of the human RNase H1 in the pharmacology of DNA-like antisense drugs. Biotechnol. ACS Nano. Part VI: Analytical and behavioural characterization of 1cyclopropanoyl d lysergic acid diethylamide (1CPLSD)", "Pharmacokinetics and subjective effects of 1P-LSD in humans after oral and intravenous administration", "Unifying Theories of Psychedelic Drug Effects", "Mammalian central nervous system trace amines. doi: 10.1128/JVI.77.14.7796-7803.2003, Hattori, Y., Hara, E., Shingu, Y., Minamiguchi, D., Nakamura, A., Arai, S., et al. ChemNanoMat 2, 220225. Science 260, 926932 (1993). doi: 10.1038/cdd.2014.134, Ulmer, J. Therefore, this suggests that physical properties of LNPs, such as particle size range and surface charge, may be considered as key screening factors to establish a down-selection workflow for in vivo gene delivery. Recent studies have reported that not only the choice of lipid components, but also the relative proportions of the lipid ingredients in the formulation, greatly influence in vivo transfection efficiency and tissue-specific delivery7,11,12,13,14. B Biointerf. Gene therapy arrives. While pre-incubation with mouse serum did not significantly impact cellular uptake level, the transfection efficiency of the best four LNPs was greatly improved; the percentage of GFP positive cells increased by 12 times after serum pre-incubation for FIII-7 (Fig. 77, 11221131. Desai, P. R., Marepally, S., Patel, A. R., Voshavar, C., Chaudhuri, A., Singh, M., et al. Such proteins could replace defective or missing proteins in the patient (Saraswat et al., 2009). doi: 10.1002/anie.201101554, Shin, H., Park, S.-J., Yim, Y., Kim, J., Choi, C., Won, C., et al. Cell Biol. Antisense Res. According to the journal Clinical Pharmacology & Therapeutics, vaporizing cannabis is a safe and more effective way (than smoking) of delivering THC to patients for medical uses. and H.-Q.M. Available online at: http://www.crisprtx.com/programs/pipeline (accessed December 27, 2019). Circ. Sci. Nanotech. Miravirsen dosing in chronic hepatitis C patients results in decreased microRNA-122 levels without affecting other microRNAs in plasma. [67][68] When VMAT2 is inhibited monoamine neurotransmitters such as dopamine cannot be released into the synapse via typical release mechanisms. The bowl and stem assembly of most bongs is removed briefly after the cannabis is burned, allowing clean air to circulate and clear the smoke chamber, ensuring no smoke dissipates without being properly consumed. As described below, this gene therapy is effective at restoring immune function in SCID. Currently, there are several cell-based therapies that have reached clinical trials and employ mRNA. Modernatx. A. Each humanized immunoglobulin chain will usually comprise, in addition to the doi: 10.1021/acsami.7b11132, Anraku, I., Harvey, T. J., Linedale, R., Gardner, J., Harrich, D., Suhrbier, A., et al. [71], Although several attempts have been made, starting in the 19th and 20th centuries, to define common phenomenological structures of the effects produced by classic psychedelics, a universally accepted taxonomy does not yet exist. Methods Clin. Bars refer to the MFI (Median fluorescence intensity) value on the left, the boxes refer to the metabolic activity on the right. A comparison of plasmid DNA and mRNA as vaccine technologies. Antisense oligonucleotides are short single-stranded DNA, phosphorothioate DNA, RNA analogs, conformationally restricted nucleosides (locked nucleic acids, LNA), or morpholino phosphorodiamidate oligonucleotides complementary to a certain region of RNA that they are meant to target. Cheng, Q. et al. Nat. 2b. Immunol. J. Med. and H.-Q.M.). Moreover, clusters that showed a moderate efficiency in vitro could have potent transfection efficiency in vivo. c, d Ex vivo imaging and quantitative luminescence measurement of the liver of BALB/c mice at 12h post-administration. doi: 10.1038/s41591-018-0327-9, Maguire, A. M., Russell, S., Wellman, J. 1, 347355. Nature 589, 189191. Rev. New Sea Serpent 22' Cuddy Cabin. (2006). Most recently, there has been a movement in nonscientific circles to recognize the ability of these substances to provoke mystical experiences and evoke feelings of spiritual significance. J. Immunol. 6). 2, pp. Based on these considerations, we did not put a limiting range for particle size or any of the other physical features, as the first set of factors for our screening workflow. Stjude (2020). doi: 10.1039/C5CS00199D, Molinaro, R., Corbo, C., Martinez, J. O., Taraballi, F., Evangelopoulos, M., Minardi, S., et al. Clinical Proof of concept for a novel hepatocyte-targeting GalNAc-siRNA conjugate. Pharm. doi: 10.1016/j.drudis.2018.02.012, Zhu, X., Yin, L., Theisen, M., Zhuo, J., Siddiqui, S., Levy, B., et al. Biomaterials 91, 7380. doi: 10.1016/j.ejpb.2013.07.002, Hanna, J., Hossain, G. S., and Kocerha, J. [75] Tryptamines are well documented to cause classic psychedelic states, such as increased empathy, visual distorsions (drifting, morphing, breathing, melting of various surfaces and objects), auditory hallucinations, ego dissolution or ego death with high enough dose, mystical and spiritual experiences, closed eye hallucinations and complete detachment from reality with a high enough dose. 15, 10371046. Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA, Yining Zhu,Ruochen Shen,Ivan Vuong,Yizong Hu,Jiayuan Kong,Sashank K. Reddy&Hai-Quan Mao, Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA, Yining Zhu,Ruochen Shen,Ivan Vuong,Zhi-Cheng Yao,Yizong Hu,Won June Cho,Jiayuan Kong,Sashank K. Reddy&Hai-Quan Mao, Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA, Yining Zhu,Ruochen Shen,Ivan Vuong,Zhi-Cheng Yao,Yizong Hu,Jiayuan Kong&Hai-Quan Mao, Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA, USA, Rebekah A. Reynolds,Melanie J. Shears&Sean C. Murphy, Center for Emerging and Re-emerging Infectious Diseases, University of Washington, Seattle, WA, USA, Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, USA, Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA, Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA, Department of Microbiology, University of Washington, Seattle, WA, USA, Seattle Malaria Clinical Trials Center, Fred Hutch Cancer Research Center, Seattle, WA, USA, You can also search for this author in