When co-delivering several antigen-encoding mRNAs, one challenge is to elicit potent specific immune responses to every antigen. 2015). 2020). For further stimulation and maturation, the DCs were transfected with mRNAs encoding specific antigens and maturation signals (Benteyn et al. The differences in the immune systems between human and other animal species may lead to distinct immune responses (Shay et al. 2016). J Immunother Cancer 7:38, Pardi N, Hogan MJ, Pelc RS et al (2017a) Zika virus protection by a single low-dose nucleoside-modified mRNA vaccination. Nat Rev Mater 2:17056, Hartmann AF, Senn MJ (1932) Studies in the metabolism of sodium r-lactate. 2016) routes. We would like to show you a description here but the site won’t allow us. Cell 170:273–83.e12, Riley RS, June CH, Langer R et al (2019) Delivery technologies for cancer immunotherapy. Mol Biol Cell 29:53–65, Reichmuth AM, Oberli MA, Jaklenec A et al (2016) mRNA vaccine delivery using lipid nanoparticles. Electroporation delivered into K562 cells two mRNAs each of which encoding half of an engineered IgG against a tumor-associated antigen, the tight-junction proteins claudin 6 (Stadler et al. Mol Ther 16:1833–1840, Kashem SW, Haniffa M, Kaplan DH (2017) Antigen-presenting cells in the skin. Proc Natl Acad Sci 109:14604–14609, Gerna G, Revello MG, Baldanti F et al (2017) The pentameric complex of human cytomegalovirus: cell tropism, virus dissemination, immune response and vaccine development. In general, the mRNA vaccines delivered by polymer materials showed therapeutic effects in preclinical studies. Proc Natl Acad Sci 110:2946, Sienkiewicz S, Palmunen JF (2017) Clinical nursing calculations, p xix, 591 p illustrations (some color); 28 cm. 2004; Li et al. Next, the mRNA-transfected DCs were validated with their phenotypes and functions, then re-introduced back to the patients to function as antigen-specific APCs (Benteyn et al. One of the three hemagglutinins encoded by the self-amplifying mRNAs failed to reduce influenza viral RNA copies and elicit adequate protection in both monovalent and trivalent formats. Therefore, clinical trials are of paramount importance for assessing the efficacy of mRNA vaccines in humans. (2019), ionizable lipid/DSPC/cholesterol/PEG-lipid: 50/10/38.5/1.5, mol/mol, Bahl et al. Moreover, a recent study reported that a cationic copolymer material co-delivering one mRNA encoding OVA and a CpG ssDNA adjuvant eliminated OVA-expressing lymphoma tumor from mice after either SC or IV administration (Haabeth et al. In the vaccination process, mRNA formulation and delivery strategies facilitate effective expression and presentation of antigens, and immune stimulation. 2007). 2015). 2014). On the other side, the activation of certain RNA sensors may inhibit mRNA translation in cell cytosol (Pardi et al. 2016; Joe et al. 1999). In this section, we focus on the technologies for formulating and delivering mRNA vaccines in carrier-mediated, naked, and DC-based forms. 2014; Alberer et al. 2020). Nat Biotechnol 37:1174–1185, Midoux P, Pichon C (2015) Lipid-based mRNA vaccine delivery systems. 2016). Journal of Materials Chemistry B 7:1824–1841, Kariko K, Buckstein M, Ni H et al (2005) Suppression of RNA recognition by Toll-like receptors: the impact of nucleoside modification and the evolutionary origin of RNA. Its vaccine efficacy in human awaits further evaluation by clinical trials. 2010; Bialkowski et al. Secondly, efficient intracellular delivery of mRNA is another challenge owing to the negative charge and large size of mRNA molecules. The advantage of VRPs arises from the efficient cytoplasmic delivery of RNA payload by viral vectors (Usme-Ciro et al. The administration route for mRNA vaccines plays an important role in determining vaccination efficacy (Eggert et al. Notably, the five subunits of hCMV PC need to be expressed and assembled into a complex by the same cell to be immunogenic (Macagno et al. Human Vaccines Immunotherapeutics 10:3146–3152, Reche P, Flower DR, Fridkis-Hareli M et al (2018) Peptide-Based Immunotherapeutics and Vaccines 2017. Vaccine 30:4341–4348, Oberli MA, Reichmuth AM, Dorkin JR et al (2016) Lipid nanoparticle assisted mRNA delivery for potent cancer immunotherapy. (2016), Gradel et al. 2014; Davis et al. These cyclic amino head groups directly bound the STING (stimulator of interferon genes) protein and triggered the downstream signaling pathway, leading to an elevated innate response. Nat Commun 8:14630, Pardi N, Hogan MJ, Porter FW et al (2018) mRNA vaccines—a new era in vaccinology. Thus, IM injection is the most widely used administration route for adjuvanted vaccines (Moyer et al. 2018). 2013). Therefore, their typical size is approximately 8–12 k nucleotides, larger than the conventional mRNA vaccine. Because of these advantages, mRNA vaccines have great potential to be manufactured and deployed in a timely manner in response to rapid infectious disease outbreaks. Researchers have investigated many methods to deliver mRNA vaccines. 2017). 2018; Kowalski et al. Adv Healthc Mater 6, Usme-Ciro JA, Campillo-Pedroza N, Almazán F et al (2013) Cytoplasmic RNA viruses as potential vehicles for the delivery of therapeutic small RNAs. Mol Ther 26:446–55, von Herrath MG, Bot A (2003) Immune responsiveness, tolerance and dsRNA: implications for traditional paradigms. acknowledges the Maximizing Investigators’ Research Award R35GM119679 from the National Institute of General Medical Sciences. Many more viral infections, bacterial infections, and various cancers were targeted using engineered VRP vaccines which were reviewed by Lundstrom (Lundstrom 2016). 2014), and in vivo in 0.8 Ringer’s lactate through intradermal (Van Lint et al. 2019b). The mechanism study suggested mRNA complex was taken up by phagocytosis and clathrin-dependent endocytosis followed by endosomal escape (Coolen et al. 2012; Fuchs et al. 2018) and immature dendritic cells (Kreiter et al. A comprehensive overview of recent advances in mRNA vaccine delivery may facilitate the future development of novel delivery strategies and effective mRNA vaccines. 2017). Methods Mol Biol 969:43–54, Weng Y, Li C, Yang T et al (2020) The challenge and prospect of mRNA therapeutics landscape. J Control Release 79:1–5, Papachristofilou A, Hipp MM, Klinkhardt U et al (2019) Phase Ib evaluation of a self-adjuvanted protamine formulated mRNA-based active cancer immunotherapy, BI1361849 (CV9202), combined with local radiation treatment in patients with stage IV non-small cell lung cancer. Similar to functional lipid-based carriers, polymers can also protect RNA from RNase-mediated degradation and facilitate intracellular delivery (Kowalski et al. Protamine is a cationic peptide used in many early studies for the delivery of mRNA vaccines. 2014). 2014). 2010; Diken et al. Among the mucosal administration routes, intranasal and intravaginal administrations were utilized to deliver mRNA vaccines (Lorenzi et al. J Nanopart Res 10:925–934, Jayaraman M, Ansell SM, Mui BL et al (2012) Maximizing the potency of siRNA lipid nanoparticles for hepatic gene silencing in vivo. : 1997) 32:498–507, Weissman D, Pardi N, Muramatsu H et al (2013) HPLC purification of in vitro transcribed long RNA. Front Immunol 9, Cullis PR, Hope MJ (2017) Lipid nanoparticle systems for enabling gene therapies. Sci Rep 6:22509, Billingsley MM, Singh N, Ravikumar P et al (2020) Ionizable lipid nanoparticle-mediated mRNA delivery for human CAR T cell engineering. Gene Ther 14:1175, Pun SH, Hoffman AS (2013) B.8—nucleic acid delivery. A short summary of this paper. 2017; Sebastian et al. Furthermore, protamine-CPP fusion protein combines cationic and cell-penetrating features. Cancer Immunol Res 7:B209, Sabnis S, Kumarasinghe ES, Salerno T et al (2018) A novel amino lipid series for mRNA delivery: improved endosomal escape and sustained pharmacology and safety in non-human primates. (2017a, b), Thran et al. Nature 547:222, Samsa MM, Dupuy LC, Beard CW et al (2019) Self-amplifying RNA vaccines for venezuelan equine encephalitis virus induce robust protective immunogenicity in mice. 2005; Fotin-Mleczek et al. 2018; Li et al. 1d). 2019). However, the total amount of the three mRNAs varied depending on specific applications and delivery routes. Cancer Res 70:9031–9040, Kubler H, Scheel B, Gnad-Vogt U et al (2015) Self-adjuvanted mRNA vaccination in advanced prostate cancer patients: a first-in-man phase I/IIa study. Подробности в лс, 'English Made Easy' Learning English through Pictures, Projects with Young Learners (Resource Books for Teachers). Even if each mRNA-encoded antigen triggers sufficient immune response when used alone, the co-delivery of several antigens may lead to competition in epitope presentation and diminished response. When delivered to the cytosol, self-amplifying mRNAs replicate themselves while expressing the designated proteins in a relatively large amount (Iavarone et al. 2010; Pardi et al. PEI was one widely used polymeric material for mRNA vaccine delivery. 2019). PEI formulations were often prepared by direct mixing PEI solution with RNA solution. Two or more independent antigen-coding mRNAs can be co-delivered to enhance and broaden immune responses. The protamine-mRNA complex is immunogenic through activation of TLR7, likely owing to its structural similarity with condensed viral RNA genome (Scheel et al. Content may be subject to copyright. 2014). Yet, multiple clinical trials for the RNActive mRNA vaccines had shown moderate efficacy, such as a weaker antibody titer than available vaccines in the clinic against rabies virus (Alberer et al. Nano Lett 20:1499–1509, Stadler CR, Bähr-Mahmud H, Celik L et al (2017) Elimination of large tumors in mice by mRNA-encoded bispecific antibodies. 2017). Mol Ther—Nucleic Acids 1:e37, Benteyn D, Heirman C, Bonehill A et al (2015) mRNA-based dendritic cell vaccines. J Control Release: Official J Control Release Soc 310:36–46, Thran M, Mukherjee J, Pönisch M et al (2017) mRNA mediates passive vaccination against infectious agents, toxins, and tumors. Select Readings Second Edition contains a range of high interest reading texts approved by experienced teachers. J Gene Med 6:1014–1022, Doyle G, McCuteheon J (2015) Intravenous medications by direct IV route. J Immunother Cancer 3:26, Leal L, Guardo AC, Morón-López S et al (2018) Phase I clinical trial of an intranodally administered mRNA-based therapeutic vaccine against HIV-1 infection. 2016; Joe et al. 2011; Tateshita et al. Vet Pathol 55:341–354, Selmi A, Vascotto F, Kautz-Neu K et al (2016) Uptake of synthetic naked RNA by skin-resident dendritic cells via macropinocytosis allows antigen expression and induction of T-cell responses in mice. in English) 55:13808–13812, Kallen K-J, Heidenreich R, Schnee M et al (2013) A novel, disruptive vaccination technology: self-adjuvanted RNActive(®) vaccines. One mRNA encoding an antigen was commonly mixed with the three mRNAs and administered simultaneously to initiate specific immunity. 2016). Vaccine 37:4204–4213, Martinon F, Krishnan S, Lenzen G et al (1993) Induction of virus-specific cytotoxic T lymphocytes in vivo by liposome-entrapped mRNA. The authors are experienced English language teachers with strong backgrounds in language analysis and language learning. Nat Med 5:823–7, Zeng C, Hou X, Yan J et al (2020) Leveraging mRNAs sequences to express SARS-CoV-2 antigens in vivo. Gene Ther 18:702–708, Dimitriadis GJ (1978) Translation of rabbit globin mRNA introduced by liposomes into mouse lymphocytes. Nat Biotechnol 31:653–658, Sahin U, Derhovanessian E, Miller M et al (2017) Personalized RNA mutanome vaccines mobilize poly-specific therapeutic immunity against cancer. In this chapter, we overview the recent progress and existing challenges in the formulation and delivery technologies of mRNA vaccines with perspectives for future development. ACS Nano 13:1655–1669, Verbeke R, Lentacker I, De Smedt SC et al (2019b) Three decades of messenger RNA vaccine development. 2018). 2014; Chahal et al. 2018). 2010; Wecker et al. J Control Release: Official J Control Release Soc 107:276–287, Hoang-Le D, Smeenk L, Anraku I et al (2009) A Kunjin replicon vector encoding granulocyte macrophage colony-stimulating factor for intra-tumoral gene therapy. 2020). 2016; Van Lint et al. In addition, DCs can be loaded with various forms of antigens and stimulatory signals and are highly amenable to such modifications (Pardi et al. The vascular and lymphatic vessels in this layer of skin also help transport mRNA vaccines and APCs to the draining lymph nodes to activate T and B cells (Kashem et al. J Control Release: Official J Control Release Soc 189:141–149, McCullough KC, Bassi I, Milona P et al (2014) Self-replicating replicon-RNA delivery to dendritic cells by chitosan-nanoparticles for translation in vitro and in vivo. 2019; Sebastian et al. Adv Mater 29:1606944, Fleeton MN, Chen M, Berglund P et al (2001) Self-replicative RNA vaccines elicit protection against influenza A virus, respiratory syncytial virus, and a tickborne encephalitis virus. 2004). J Immunother 34:1–15, Fotin-Mleczek M, Zanzinger K, Heidenreich R et al (2014) mRNA-based vaccines synergize with radiation therapy to eradicate established tumors. 2016; Sedic et al. 2016; Van Lint et al. The mouse group immunized with two co-delivered self-amplifying mRNAs showed similar immune response and protection as the group received the nucleoprotein self-amplifying mRNA alone. 2018). PLoS ONE 13:e0201464, Belliveau NM, Huft J, Lin PJC et al (2012) Microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of siRNA. (Gurpreet and Singh 2018). However, mRNA was translated poorly when in complex with protamine (Scheel et al. 2012; Fuchs et al. 2012; Schnee et al. : HP37-25F-PDF ISBN: 2371-5383 Pub. Intravaginal injection is another approach to deliver mRNA vaccines to the site of infection to express neutralizing antibodies. 2011). 2019). 2017), ethanol injection (Geall et al. 2017). (2001), Moyer et al. For example, delivery carriers, such as lipid-derived and polymer-derived materials, dramatically increased cellular uptake of RNAs, thus receiving tremendous attention in recent years (Reichmuth et al. Additionally, the lymph vessels may directly transport small-sized lipid nanoparticles to draining lymph nodes (Moyer et al. Important electrical characteristics, such as voltage, capacitance, and resistance, were adjusted to improve the delivery efficiency (Van Tendeloo et al. Similar to the nasal mucosa and NALT, the immature and activated APCs available in the lung can engulf and process the mRNA vaccine (Stehle et al. Mechanism of action studies suggested the GALA peptide facilitated the cell uptake and release of mRNA into the cytosol through binding to sialic acid groups on the DC surface (Lou et al. The results indicated that the hydrophilic head group in lipid materials might determine the acid dissociation constant (pKa) and influence the delivery efficiency (Sato et al. In one report, the intravaginal delivery of the mRNA vaccine encoding an anti-HIV antibody induced high levels of antibody expression in the reproductive tract of sheep and rhesus macaques (Lindsay et al. 2017). Various delivery strategies have been investigated to address these obstacles with different delivery materials, formulation methods, and routes of administrations. 2017). 2018). The other feature of the naked mRNA vaccine, especially those made of unmodified nucleotides, is its intrinsic immunogenicity, which serves as a double-edged sword. To increase injection volume and mRNA dose, patients received multiple ID injections at different sites per visit in a clinical trial (Sebastian et al. A similar observation was reported in another study in 2018 (Vogel et al. The mRNA dose necessary to induce sufficient immune response in mice and other animals might not be directly correlated to humans. Intranasal delivery can also apply mRNA to the lung through the trachea. After SC injection, the expressed antigen was detected in DCs (McCullough et al. 2004, 2005), limiting the expression of the encoded antigen and reducing its potential as an independent mRNA carrier. Exp Dermatol 19:19–28, Hajj KA, Whitehead KA (2017) Tools for translation: non-viral materials for therapeutic mRNA delivery.