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Nanoparticle‐Mediated DNA and Mrna Delivery REVIEW www.advhealthmat.de Next-Generation Vaccines: Nanoparticle-Mediated DNA and mRNA Delivery William Ho, Mingzhu Gao, Fengqiao Li, Zhongyu Li, Xue-Qing Zhang,* and Xiaoyang Xu* organism-based vaccines have wiped out or Nucleic acid vaccines are a method of immunization aiming to elicit immune nearly eradicated many once great killers responses akin to live attenuated vaccines. In this method, DNA or messenger of humanity, including smallpox, polio, RNA (mRNA) sequences are delivered to the body to generate proteins, which measles, mumps, rubella, diphtheria, per- [1–4] mimic disease antigens to stimulate the immune response. Advantages of tussis, and tetanus. However, the quick emergence of diseases such as SARS- nucleic acid vaccines include stimulation of both cell-mediated and humoral CoV-2, H1N1 as well as quickly evolving immunity, ease of design, rapid adaptability to changing pathogen strains, and deadly diseases like Ebola create a challenge customizable multiantigen vaccines. To combat the SARS-CoV-2 pandemic, for conventional vaccines such as live at- and many other diseases, nucleic acid vaccines appear to be a promising tenuated viral vaccines (LAV), and inacti- [5,6] method. However, aid is needed in delivering the fragile DNA/mRNA payload. vated/killed viral vaccines, which with Many delivery strategies have been developed to elicit effective immune the traditional vaccine development path- way may take on average over 10 years to stimulation, yet no nucleic acid vaccine has been FDA-approved for human develop, or with Ebola requiring an acceler- use. Nanoparticles (NPs) are one of the top candidates to mediate successful ated 5-year development,[7] and even more DNA/mRNA vaccine delivery due to their unique properties, including time needed to scale up manufacturing and unlimited possibilities for formulations, protective capacity, simultaneous stockpile for a large country. As a result, loading, and delivery potential of multiple DNA/mRNA vaccines. This review during the vaccine development, scale-up, and implementation many get sick and lose will summarize the many varieties of novel NP formulations for DNA and their lives. The most recent and relevant mRNA vaccine delivery as well as give the reader a brief synopsis of NP case for discussion is the ongoing SARS- vaccine clinical trials. Finally, the future perspectives and challenges for CoV-2 pandemic, which is considered one NP-mediated nucleic acid vaccines will be explored. of the most crucial global health calami- ties of the century. According to the report from the World Health Organization (WHO as of December 6, 2020), the current outbreak of SARS-CoV-2 1. Introduction has caused over 1.54 million deaths in more than 200 countries throughout the world, as well as a worldwide economic shutdown Since the conceptualization of vaccines and their implementa- of many countries leading to widespread disorder, continental tion on a governmental scale, live attenuated or killed whole lockdowns and fiscal uncertainty. With pandemics certain to re-emerge in the future, a modern- ized vaccination system must be developed to ensure that future W. Ho, F. Li, Z. Li, Prof. X. Xu pandemics are controlled rapidly with minimal loss of life and Department of Chemical and Materials Engineering New Jersey Institute of Technology minimal disruption of the economy. Conventional seasonal an- Newark, NJ 07102, USA nual vaccine formulations often fail to match strains in circula- E-mail: [email protected] tion due to antigenic drift of viral strains, especially when they oc- M. Gao, Prof. X.-Q. Zhang cur late in the flu season.[8–10] Vaccines for new viral strains like Engineering Research Center of Cell & Therapeutic Antibody SARS-CoV-2 are time-consuming to create and distribute; iden- Ministry of Education and School of Pharmacy tifying the virus, developing, testing, obtaining approval from Shanghai Jiao Tong University regulatory agencies and mass production take at least several 800 Dongchuan Road Shanghai 200240, P. R. China months to years, as evidenced in the 2009 H1N1 pandemic as E-mail: [email protected] well as the 2014–2016 outbreaks of Ebola. It is clear that a new Prof. X. Xu model of vaccine production should emerge to tackle this urgent Department of Biomedical Engineering problem. In the past decades, there have been attempts to replace New Jersey Institute of Technology 323 Dr Martin Luther King Jr Blvd Newark, NJ 07102, USA inactivated or live attenuated vaccines, through development of modern vaccine technologies such as virus-like particles, peptide- The ORCID identification number(s) for the author(s) of this article based vaccines, as well as nucleic acid-based vaccines.[11] These can be found under https://doi.org/10.1002/adhm.202001812 newer developments aimed to improve vaccine stability, safety, DOI: 10.1002/adhm.202001812 and cost.[12] Nucleic acid vaccines in particular are a recent and Adv. Healthcare Mater. 2021, 2001812 2001812 (1 of 17) © 2021 Wiley-VCH GmbH www.advancedsciencenews.com www.advhealthmat.de cost-effective development in the biomedical field which have at- 2. DNA and mRNA Present Many Advantages for tracted great amounts of attention. DNA and mRNA vaccines aim Vaccine Development to use host cell machinery to produce coded protein antigens, stimulating humoral and cell-mediated immunity through pro- DNA is the genetic material located within the nucleus, while duction of neutralizing antibodies and cytotoxic T lymphocytes mRNA is the intermediate which ferries the information from (CTLs). Therefore nucleic acid vaccines may be the key to an ef- the nuclear DNA to be translated into functional proteins in the fective vaccine, as they offer quick turnaround utilizing generic cytoplasm. They both participate in the cellular protein transla- DNA/mRNA manufacturing processes, ease of sequence mod- tion pathway. The double stranded DNA represents the gene it- ification to adapt to changing pathogen strains, and the ability self and is located in the nucleus, whereas single stranded mRNA to elicit both antibody and cytotoxic T-lymphocyte responses.[13] is the transcribed version of the gene which associates with the Further advantages of DNA/mRNA vaccines include the absence ribosomes to translate codons for protein production. Nucleic of living or killed organisms in the vaccine and the ability to acid vaccines have many advantages: they avoid issues associ- specifically direct immune responses toward only the coded anti- ated with recombinant protein vaccines such as improper pro- gens in the vaccine. tein folding or high protein purification cost, they do not display Additionally, administration of nucleic acid vaccines gener- the infectious risks associated with attenuated or inactivated vac- ates endogenous proteins displaying the native conformation cines produced from live infectious organisms, and they can acti- with posttranslational modifications like those found in natural vate both the humoral and cellular immune response, leading to pathogen infection.[14] Although plasmid DNA and mRNA vastly improved protective immune responses.[15] Furthermore, vaccines display these myriad advantages, and have been widely improved transfection efficiency and stability of mRNA can be evaluated in clinical trials, so far none have been licensed for achieved through various chemical modifications, leading to en- human use and it seems as of now current therapies fail to har- hanced immunity. Plasmid DNA (pDNA) sequences contain a ness the full potential of this promising therapeutic strategy.[5,15] promoter region, an intron, antigen sequence, and a polyA sig- This is largely because efficient delivery to cells remains elusive. nal. mRNAs are usually produced by in vitro transcription of a In the human body, nucleic acids are fragile and are degraded cDNA template such as pDNA with RNA polymerase.[16] Syn- rapidly due to endogenous nucleases. DNA/mRNA must also thetic mRNA comprises a protein-encoding open reading frame cross many cellular barriers to reach the cytoplasm (for mRNA (ORF) with a 5’ cap,” and a 3’ poly(A) tail. 5′ and 3′ untrans- vaccines) and nucleus (in the case of DNA vaccines). As a result, lated regions (UTRs) flanking the ORF will increase the transla- the immunogenicity of the nucleic acid vaccines which have tion and stability.[17] Chemical modifications may be made to the been developed is low. Finally, due to these reasons, the delivery DNA and mRNA to improve transfection and stability, including of DNA and mRNA vaccines has yet to be clinically validated in codon optimization, prefusion stabilization mutations and addi- humans through Phase III.[1,2] tions to support trimerization[18,19] Additionally, for DNA, pro- As viral vector delivery has the risk of increasing public moter selection and modifying the plasmid backbone (i.e., re- health concerns which may hinder widespread adoption, non- moving bacterial elements) can play a large role in increasing viral methods of gene delivery are being increasingly explored. the gene expression.[19–23] Finally, for mRNA the optimization of Nanoparticle-mediated delivery is one such nonviral delivery the 5’- and 3’- UTRs have been utilized to improve translational method which holds great promise for efficient delivery of efficacy[17,24] and the incorporation of modified nucleosides such nucleic acids and may represent the future of next generation as pseudouridine (Ψ) and 5-methylcytidine (5 mC) have been vaccines. NPs possess many advantages to facilitate vaccine de- shown to reduce immune recognition of the mRNA.[1,25–27] Self- livery; they protect the nucleic acid payload from degradation and amplifying mRNA has been recently developed as another nu- offer versatile formulation strategies with a variety of biomaterial cleic acid-based vaccine technology. The self-amplifying mRNAs options to overcome the barriers of cell internalization, improve are termed replicons, and are derived from RNA viruses where specific immune cell targeting through surface modifications the structural viral proteins are replaced with mRNA encoding and may utilize pH-sensitive materials to enhance endosomal antigens and RNA polymerases.
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