{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T07:27:42Z","timestamp":1790753262701,"version":"4.1.0"},"reference-count":67,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,2,10]],"date-time":"2021-02-10T00:00:00Z","timestamp":1612915200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>Vaccination is the most effective method to date to prevent viral diseases. It intends to mimic a naturally occurring infection while avoiding the disease, exposing our bodies to viral antigens to trigger an immune response that will protect us from future infections. Among different strategies for vaccine development, recombinant vaccines are one of the most efficient ones. Recombinant vaccines use safe viral vectors as vehicles and incorporate a transgenic antigen of the pathogen against which we intend to generate an immune response. These vaccines can be based on replication-deficient viruses or replication-competent viruses. While the most effective strategy involves replication-competent viruses, they must be attenuated to prevent any health hazard while guaranteeing a strong humoral and cellular immune response. Several attenuation strategies for adenoviral-based vaccine development have been contemplated over time. In this paper, we will review them and discuss novel approaches based on the principle that protein synthesis from individual genes can be modulated by codon usage bias manipulation. We will summarize vaccine approaches that consider recoding of viral proteins to produce adenoviral attenuation and recoding of the transgene antigens for both viral attenuation and efficient viral epitope expression.<\/jats:p>","DOI":"10.3389\/fmicb.2021.633946","type":"journal-article","created":{"date-parts":[[2021,2,10]],"date-time":"2021-02-10T17:29:38Z","timestamp":1612978178000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":21,"title":["Codon Usage and Adenovirus Fitness: Implications for Vaccine Development"],"prefix":"10.3389","volume":"12","author":[{"given":"Judit","family":"Gim\u00e9nez-Roig","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Estela","family":"N\u00fa\u00f1ez-Manch\u00f3n","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramon","family":"Alemany","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eneko","family":"Villanueva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Fillat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,2,10]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1111\/imr.12861","article-title":"Factors in B cell competition and immunodominance.","volume":"296","author":"Abbott","year":"2020","journal-title":"Immunol. Rev."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0031177","article-title":"Pre-clinical evaluation of a replication-competent recombinant adenovirus serotype 4 vaccine expressing influenza H5 hemagglutinin.","volume":"7","author":"Alexander","year":"2012","journal-title":"PLoS One"},{"key":"B3","doi-asserted-by":"publisher","first-page":"4527","DOI":"10.1093\/nar\/gku075","article-title":"The influence of CpG and UpA dinucleotide frequencies on RNA virus replication and characterization of the innate cellular pathways underlying virus attenuation and enhanced replication.","volume":"42","author":"Atkinson","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"B4","doi-asserted-by":"publisher","first-page":"320","DOI":"10.1126\/science.aab3886","article-title":"Protective efficacy of adenovirus\/protein vaccines against SIV challenges in rhesus monkeys.","volume":"349","author":"Barouch","year":"2015","journal-title":"Science"},{"key":"B5","doi-asserted-by":"publisher","first-page":"6179","DOI":"10.18632\/oncotarget.3350","article-title":"Late-phase miRNA-controlled oncolytic adenovirus for selective killing of cancer cells.","volume":"6","author":"Bofill-De Ros","year":"2015","journal-title":"Oncotarget"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21144821","article-title":"Non-Human Primate-Derived Adenoviruses for Future Use as Oncolytic Agents?","volume":"21","author":"Bots","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B7","doi-asserted-by":"publisher","first-page":"2069","DOI":"10.7205\/MILMED-D-17-00109","article-title":"Adenovirus 4\/7 Vaccine\u2019s Effect on Disease Rates Is Associated With Disappearance of Adenovirus on Building Surfaces at a Military Recruit Base.","volume":"182","author":"Broderick","year":"2017","journal-title":"Mil. Med."},{"key":"B8","doi-asserted-by":"publisher","first-page":"2744","DOI":"10.7150\/thno.40902","article-title":"Oligopeptide-modified poly(beta-amino ester)s-coated AdNuPARmE1A: Boosting the efficacy of intravenously administered therapeutic adenoviruses.","volume":"10","author":"Brugada-Vil\u00e0","year":"2020","journal-title":"Theranostics"},{"key":"B9","doi-asserted-by":"publisher","first-page":"283","DOI":"10.1016\/j.tig.2017.02.001","article-title":"Synonymous Codons: Choose Wisely for Expression.","volume":"33","author":"Brule","year":"2017","journal-title":"Trends Genet."},{"key":"B10","doi-asserted-by":"publisher","first-page":"341","DOI":"10.1016\/j.molcel.2016.01.008","article-title":"Synonymous Codons Direct Cotranslational Folding toward Different Protein Conformations.","volume":"61","author":"Buhr","year":"2016","journal-title":"Mol. Cell"},{"key":"B11","doi-asserted-by":"publisher","first-page":"9957","DOI":"10.1128\/JVI.00508-09","article-title":"Genetic inactivation of poliovirus infectivity by increasing the frequencies of CpG and UpA dinucleotides within and across synonymous capsid region codons.","volume":"83","author":"Burns","year":"2009","journal-title":"J. Virol."},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00039-20","article-title":"A Lassa Fever Live-Attenuated Vaccine Based on Codon Deoptimization of the Viral Glycoprotein Gene.","volume":"11","author":"Cai","year":"2020","journal-title":"MBio"},{"key":"B13","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1038\/nrg1770","article-title":"Hearing silence: non-neutral evolution at synonymous sites in mammals.","volume":"7","author":"Chamary","year":"2006","journal-title":"Nat. Rev. Genet."},{"key":"B14","doi-asserted-by":"publisher","first-page":"3531","DOI":"10.1002\/1873-3468.13695","article-title":"Viral and cellular interactions during adenovirus DNA replication.","volume":"593","author":"Charman","year":"2019","journal-title":"FEBS Lett."},{"key":"B15","doi-asserted-by":"publisher","first-page":"1784","DOI":"10.1126\/science.1155761","article-title":"Virus attenuation by genome-scale changes in codon pair bias.","volume":"320","author":"Coleman","year":"2008","journal-title":"Science"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.00909","article-title":"Factors Which Contribute to the Immunogenicity of Non-replicating Adenoviral Vectored Vaccines.","volume":"11","author":"Coughlan","year":"2020","journal-title":"Front. Immunol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"669","DOI":"10.1128\/JVI.02184-14","article-title":"Amplified and persistent immune responses generated by single-cycle replicating adenovirus vaccines.","volume":"89","author":"Crosby","year":"2015","journal-title":"J. Virol."},{"key":"B18","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1093\/dnares\/12.2.91","article-title":"Codon and amino acid usage in two major human pathogens of genus Bartonella\u2013optimization between replicational-transcriptional selection, translational control and cost minimization.","volume":"12","author":"Das","year":"2005","journal-title":"DNA Res."},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-37249-4","article-title":"Molecular Evolution of Human Adenovirus (HAdV) Species C.","volume":"9","author":"Dhingra","year":"2019","journal-title":"Sci. Rep."},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-020-0849-9","article-title":"Deep splicing plasticity of the human adenovirus type 5 transcriptome drives virus evolution.","volume":"3","author":"Donovan-Banfield","year":"2020","journal-title":"Commun. Biol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1016\/s0959-437x(02)00353-2","article-title":"Evolution of synonymous codon usage in metazoans.","volume":"12","author":"Duret","year":"2002","journal-title":"Curr. Opin. Genet. Dev."},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-18077-5","article-title":"An adenovirus-vectored COVID-19 vaccine confers protection from SARS-COV-2 challenge in rhesus macaques.","volume":"11","author":"Feng","year":"2020","journal-title":"Nat. Commun."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1038\/s41426-018-0102-5","article-title":"An adenovirus serotype 2-vectored ebolavirus vaccine generates robust antibody and cell-mediated immune responses in mice and rhesus macaques.","volume":"7","author":"Feng","year":"2018","journal-title":"Emerg. Microbes Infect."},{"key":"B24","doi-asserted-by":"publisher","first-page":"467","DOI":"10.1016\/S0140-6736(20)31604-4","article-title":"Safety and immunogenicity of the ChAdOx1 nCoV-19 vaccine against SARS-CoV-2: a preliminary report of a phase 1\/2, single-blind, randomised controlled trial.","volume":"396","author":"Folegatti","year":"2020","journal-title":"Lancet"},{"key":"B25","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1172\/JCI60202","article-title":"Human adenovirus-specific T cells modulate HIV-specific T cell responses to an Ad5-vectored HIV-1 vaccine.","volume":"122","author":"Frahm","year":"2012","journal-title":"J. Clin. Invest."},{"key":"B26","doi-asserted-by":"publisher","first-page":"679","DOI":"10.1016\/j.cell.2016.05.070","article-title":"Adjacent Codons Act in Concert to Modulate Translation Efficiency in Yeast.","volume":"166","author":"Gamble","year":"2016","journal-title":"Cell"},{"key":"B27","doi-asserted-by":"publisher","first-page":"3609","DOI":"10.1002\/1873-3468.13691","article-title":"State-of-the-art human adenovirus vectorology for therapeutic approaches.","volume":"593","author":"Gao","year":"2019","journal-title":"FEBS Lett."},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines8020293","article-title":"Prospects of Replication-Deficient Adenovirus Based Vaccine Development against SARS-CoV-2.","volume":"8","author":"Garofalo","year":"2020","journal-title":"Vaccines"},{"key":"B29","doi-asserted-by":"publisher","first-page":"r49","DOI":"10.1093\/nar\/8.1.197-c","article-title":"Codon catalog usage and the genome hypothesis.","volume":"8","author":"Grantham","year":"1980","journal-title":"Nucleic Acids Res."},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.107586","article-title":"Mechanism of Virus Attenuation by Codon Pair Deoptimization.","volume":"31","author":"Groenke","year":"2020","journal-title":"Cell Rep."},{"key":"B31","doi-asserted-by":"publisher","first-page":"238","DOI":"10.1016\/S1473-3099(12)70345-6","article-title":"Safety and immunogenicity of an oral, replicating adenovirus serotype 4 vector vaccine for H5N1 influenza: a randomised, double-blind, placebo-controlled, phase 1 study.","volume":"13","author":"Gurwith","year":"2013","journal-title":"Lancet Infect. Dis."},{"key":"B32","doi-asserted-by":"publisher","first-page":"3699","DOI":"10.1073\/pnas.86.10.3699","article-title":"Nonrandom utilization of codon pairs in Escherichia coli.","volume":"86","author":"Gutman","year":"1989","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B33","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1038\/sj.mt.6300024","article-title":"Type I interferon inhibits antibody responses induced by a chimpanzee adenovirus vector.","volume":"15","author":"Hensley","year":"2007","journal-title":"Mol. Ther."},{"key":"B34","doi-asserted-by":"publisher","first-page":"287","DOI":"10.1146\/annurev.genet.42.110807.091442","article-title":"Selection on codon bias.","volume":"42","author":"Hershberg","year":"2008","journal-title":"Annu. Rev. Genet."},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.15252\/embr.201948220","article-title":"Codon bias confers stability to human mRNAs.","volume":"20","author":"Hia","year":"2019","journal-title":"EMBO Rep."},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1186\/s42490-019-0016-x","article-title":"Overcoming the limitations of locally administered oncolytic virotherapy.","volume":"1","author":"Hong","year":"2019","journal-title":"BMC Biomed. Eng."},{"key":"B37","doi-asserted-by":"publisher","first-page":"1077","DOI":"10.1056\/NEJM195412302512701","article-title":"Adenoidal-pharyngeal-conjunctival agents: a newly recognized group of common viruses of the respiratory system.","volume":"251","author":"Huebner","year":"1954","journal-title":"N. Engl. J. Med."},{"key":"B38","doi-asserted-by":"publisher","first-page":"2957","DOI":"10.1534\/g3.117.041020","article-title":"Reduced Protein Expression in a Virus Attenuated by Codon Deoptimization.","volume":"7","author":"Jack","year":"2017","journal-title":"G3"},{"key":"B39","doi-asserted-by":"publisher","first-page":"648","DOI":"10.1002\/jgm.1629","article-title":"High-capacity adenoviral vectors circumvent the limitations of \u0394E1 and \u0394E1\/\u0394E3 adenovirus vectors to induce multispecific transgene product-directed CD8 T-cell responses.","volume":"13","author":"Kron","year":"2011","journal-title":"J. Gene Med."},{"key":"B40","doi-asserted-by":"publisher","first-page":"2963","DOI":"10.1016\/j.vaccine.2013.04.035","article-title":"A phase 3, randomized, double-blind, placebo-controlled study of the safety and efficacy of the live, oral adenovirus type 4 and type 7 vaccine, in U.S. military recruits.","volume":"31","author":"Kuschner","year":"2013","journal-title":"Vaccine"},{"key":"B41","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1586\/14760584.2016.1124762","article-title":"Molecular mechanisms for enhanced DNA vaccine immunogenicity.","volume":"15","author":"Li","year":"2016","journal-title":"Expert Rev. Vaccines"},{"key":"B42","doi-asserted-by":"publisher","first-page":"441","DOI":"10.1128\/CMR.00116-13","article-title":"Adenovirus infections in immunocompetent and immunocompromised patients.","volume":"27","author":"Lion","year":"2014","journal-title":"Clin. Microbiol. Rev."},{"key":"B43","doi-asserted-by":"publisher","first-page":"887","DOI":"10.1016\/S0140-6736(20)31866-3","article-title":"Safety and immunogenicity of an rAd26 and rAd5 vector-based heterologous prime-boost COVID-19 vaccine in two formulations: two open, non-randomised phase 1\/2 studies from Russia.","volume":"396","author":"Logunov","year":"2020","journal-title":"Lancet"},{"key":"B44","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1089\/hum.2013.235","article-title":"Adenovirus-based vaccines for fighting infectious diseases and cancer: progress in the field.","volume":"25","author":"Majhen","year":"2014","journal-title":"Hum. Gene Ther."},{"key":"B45","doi-asserted-by":"publisher","first-page":"10506","DOI":"10.1093\/nar\/gkz831","article-title":"Synonymous genome recoding: a tool to explore microbial biology and new therapeutic strategies.","volume":"47","author":"Mart\u00ednez","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"B46","doi-asserted-by":"publisher","first-page":"1166","DOI":"10.1128\/CVI.00510-15","article-title":"Attenuation of Replication-Competent Adenovirus Serotype 26 Vaccines by Vectorization.","volume":"22","author":"Maxfield","year":"2015","journal-title":"Clin. Vaccine Immunol."},{"key":"B47","doi-asserted-by":"publisher","first-page":"583","DOI":"10.1038\/s41586-020-2607-z","article-title":"Single-shot Ad26 vaccine protects against SARS-CoV-2 in rhesus macaques.","volume":"586","author":"Mercado","year":"2020","journal-title":"Nature"},{"key":"B48","doi-asserted-by":"publisher","first-page":"9687","DOI":"10.1128\/JVI.00738-06","article-title":"Reduction of the rate of poliovirus protein synthesis through large-scale codon deoptimization causes attenuation of viral virulence by lowering specific infectivity.","volume":"80","author":"Mueller","year":"2006","journal-title":"J. Virol."},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1101\/2020.06.19.161026","article-title":"Transgene codon usage drives viral fitness and therapeutic efficacy in oncolytic adenoviruses.","volume":"2020","author":"N\u00fa\u00f1ez-Manch\u00f3n","year":"2020","journal-title":"bioRxiv"},{"key":"B50","doi-asserted-by":"publisher","first-page":"929","DOI":"10.1093\/nar\/22.6.929","article-title":"Divergence in codon usage of Lactobacillus species.","volume":"22","author":"Pouwels","year":"1994","journal-title":"Nucleic Acids Res."},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1002603","article-title":"Balanced codon usage optimizes eukaryotic translational efficiency.","volume":"8","author":"Qian","year":"2012","journal-title":"PLoS Genet."},{"key":"B52","doi-asserted-by":"publisher","first-page":"1129","DOI":"10.1172\/JCI78280","article-title":"Antigen expression determines adenoviral vaccine potency independent of IFN and STING signaling.","volume":"125","author":"Quinn","year":"2015","journal-title":"J. Clin. Invest."},{"key":"B53","doi-asserted-by":"publisher","first-page":"546","DOI":"10.1016\/j.copbio.2007.10.010","article-title":"Replicating and non-replicating viral vectors for vaccine development.","volume":"18","author":"Robert-Guroff","year":"2007","journal-title":"Curr. Opin. Biotechnol."},{"key":"B54","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1016\/j.molimm.2018.08.011","article-title":"Selection of immunodominant epitopes during antigen processing is hierarchical.","volume":"113","author":"Sadegh-Nasseri","year":"2019","journal-title":"Mol. Immunol."},{"key":"B55","doi-asserted-by":"publisher","first-page":"1609","DOI":"10.1038\/mt.2008.141","article-title":"The immunogenicity of adenovirus vectors limits the multispecificity of CD8 T-cell responses to vector-encoded transgenic antigens.","volume":"16","author":"Schirmbeck","year":"2008","journal-title":"Mol. Ther."},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01184-17","article-title":"Immunodominance of Adenovirus-Derived CD8(+) T Cell Epitopes Interferes with the Induction of Transgene-Specific Immunity in Adenovirus-Based Immunization.","volume":"91","author":"Sch\u00f6ne","year":"2017","journal-title":"J. Virol."},{"key":"B57","doi-asserted-by":"publisher","first-page":"5701","DOI":"10.1128\/JVI.00354-11","article-title":"Using the whole-genome sequence to characterize and name human adenoviruses.","volume":"85","author":"Seto","year":"2011","journal-title":"J. Virol."},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0030302","article-title":"Immunogenic comparison of chimeric adenovirus 5\/35 vector carrying optimized human immunodeficiency virus clade C genes and various promoters.","volume":"7","author":"Shoji","year":"2012","journal-title":"PLoS One"},{"key":"B59","doi-asserted-by":"publisher","first-page":"E3633","DOI":"10.1073\/pnas.1507339112","article-title":"Attenuation of dengue (and other RNA viruses) with codon pair recoding can be explained by increased CpG\/UpA dinucleotide frequencies.","volume":"112","author":"Simmonds","year":"2015","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B60","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2012.00068","article-title":"Louis pasteur, the father of immunology?","volume":"3","author":"Smith","year":"2012","journal-title":"Front. Immunol."},{"key":"B61","doi-asserted-by":"publisher","first-page":"616","DOI":"10.1016\/j.ymthe.2004.07.013","article-title":"Adenoviruses as vaccine vectors.","volume":"10","author":"Tatsis","year":"2004","journal-title":"Mol. Ther."},{"key":"B62","doi-asserted-by":"publisher","DOI":"10.1186\/s12967-016-1042-2","article-title":"Discovery of immunodominant T-cell epitopes reveals penton protein as a second immunodominant target in human adenovirus infection.","volume":"14","author":"Tischer","year":"2016","journal-title":"J. Transl. Med."},{"key":"B63","doi-asserted-by":"publisher","DOI":"10.1038\/srep27546","article-title":"Codon optimization of the adenoviral fiber negatively impacts structural protein expression and viral fitness.","volume":"6","author":"Villanueva","year":"2016","journal-title":"Sci. Rep."},{"key":"B64","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines6030081","article-title":"Progress in Adenoviral Capsid-Display Vaccines.","volume":"6","author":"Vujadinovic","year":"2018","journal-title":"Biomedicines"},{"key":"B65","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-17972-1","article-title":"A single dose of an adenovirus-vectored vaccine provides protection against SARS-CoV-2 challenge.","volume":"11","author":"Wu","year":"2020","journal-title":"Nat. Commun."},{"key":"B66","doi-asserted-by":"publisher","first-page":"4972","DOI":"10.1128\/JVI.73.6.4972-4982.1999","article-title":"Papillomavirus capsid protein expression level depends on the match between codon usage and tRNA availability.","volume":"73","author":"Zhou","year":"1999","journal-title":"J. Virol."},{"key":"B67","doi-asserted-by":"publisher","first-page":"479","DOI":"10.1016\/S0140-6736(20)31605-6","article-title":"Immunogenicity and safety of a recombinant adenovirus type-5-vectored COVID-19 vaccine in healthy adults aged 18 years or older: a randomised, double-blind, placebo-controlled, phase 2 trial.","volume":"396","author":"Zhu","year":"2020","journal-title":"Lancet"}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2021.633946\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,2,10]],"date-time":"2021-02-10T17:29:40Z","timestamp":1612978180000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2021.633946\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,2,10]]},"references-count":67,"alternative-id":["10.3389\/fmicb.2021.633946"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2021.633946","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,2,10]]},"article-number":"633946"}}