{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:29:08Z","timestamp":1760146148696,"version":"build-2065373602"},"reference-count":46,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2024,10,3]],"date-time":"2024-10-03T00:00:00Z","timestamp":1727913600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI","award":["JP 17K08464"],"award-info":[{"award-number":["JP 17K08464"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Numerous gene therapy drugs for cancer have received global approval, yet their efficacy against solid tumors remains inadequate. Our previous research indicated that the fiber protein, a component of the adenovirus capsid, can propagate from infected cells to neighboring cells that express the adenovirus receptor. We hypothesize that merging this fiber protein with an anti-cancer protein could enable the anti-cancer protein to disseminate around the transfected cells, presenting a novel approach to cancer gene therapy. In our study, we discovered that the knob region of the adenovirus type 5 fiber protein is the smallest unit capable of spreading to adjacent cells in a receptor-specific manner. We also showed that the recombinant knob protein infiltrates cells after dispersing to surrounding cells. To assess the potential of the knob protein to augment gene therapy for solid tumors in mice, we expressed a fusion gene of the A subunit of cytotoxic cholera toxin and the knob region in mouse tumors. We found that this fusion protein only inhibited tumor growth in receptor-expressing mouse melanomas, and this inhibitory effect persisted even in mice with anti-knob antibodies. Our study\u2019s findings propose a novel cancer gene therapy strategy that enhances therapeutic effects by specifically delivering therapeutic proteins, expressed from in vivo administered genes, to target molecules. This outcome offers a fresh perspective on gene therapy for solid cancers, and we anticipate that knob proteins will serve as a platform for this method.<\/jats:p>","DOI":"10.3390\/ijms251910679","type":"journal-article","created":{"date-parts":[[2024,10,3]],"date-time":"2024-10-03T10:31:13Z","timestamp":1727951473000},"page":"10679","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Utilizing Adenovirus Knob Proteins as Carriers in Cancer Gene Therapy Amidst the Presence of Anti-Knob Antibodies"],"prefix":"10.3390","volume":"25","author":[{"given":"Naoya","family":"Koizumi","sequence":"first","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takamasa","family":"Hirai","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"},{"name":"Division of Cell-Based Therapeutic Products, National Institute of Health Sciences, Kawasaki 210-9501, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junpei","family":"Kano","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna","family":"Sato","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yurika","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arisa","family":"Sasaki","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tetsuya","family":"Nomura","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naoki","family":"Utoguchi","sequence":"additional","affiliation":[{"name":"Laboratory of Pharmaceutics and Biopharmaceutics, Showa Pharmaceutical University, Tokyo 194-8543, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3425291","DOI":"10.1155\/2019\/3425291","article-title":"Efficiency of CAR-T Therapy for Treatment of Solid Tumor in Clinical Trials: A Meta-Analysis","volume":"2019","author":"Hou","year":"2019","journal-title":"Dis. Markers"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"216871","DOI":"10.1016\/j.canlet.2024.216871","article-title":"The dilemmas and possible solutions for CAR-T cell therapy application in solid tumors","volume":"591","author":"Wang","year":"2024","journal-title":"Cancer Lett."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Rahman, M.M., and McFadden, G. (2021). Oncolytic Viruses: Newest Frontier for Cancer Immunotherapy. Cancers, 13.","DOI":"10.3390\/cancers13215452"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.omto.2021.05.002","article-title":"Current Strategies in Engaging Oncolytic Viruses with Antitumor Immunity","volume":"22","author":"Boagni","year":"2021","journal-title":"Mol. Ther. Oncolytics"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"950","DOI":"10.1016\/j.vaccine.2009.10.145","article-title":"International Epidemiology of Human Pre-Existing Adenovirus (Ad) Type-5, Type-6, Type-26 and Type-36 Neutralizing Antibodies: Correlates of High Ad5 Titers and Implications for Potential HIV Vaccine Trials","volume":"28","author":"Mast","year":"2010","journal-title":"Vaccine"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"8263","DOI":"10.1128\/JVI.77.15.8263-8271.2003","article-title":"Replication-Deficient Human Adenovirus Type 35 Vectors for Gene Transfer and Vaccination: Efficient Human Cell Infection and Bypass of Preexisting Adenovirus Immunity","volume":"77","author":"Vogels","year":"2003","journal-title":"J. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1016\/S0042-6822(03)00161-2","article-title":"Development of Adenovirus Serotype 35 as a Gene Transfer Vector","volume":"311","author":"Ganesh","year":"2003","journal-title":"Virology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5203","DOI":"10.1016\/j.vaccine.2011.05.025","article-title":"International Seroepidemiology of Adenovirus Serotypes 5, 26, 35, and 48 in Pediatric and Adult Populations","volume":"29","author":"Barouch","year":"2011","journal-title":"Vaccine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3837","DOI":"10.1016\/j.vaccine.2011.03.042","article-title":"Epidemiology of Adenovirus Type 5 Neutralizing Antibodies in Healthy People and AIDS Patients in Guangzhou, Southern China","volume":"29","author":"Sun","year":"2011","journal-title":"Vaccine"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1128\/JVI.01878-08","article-title":"Effect of Neutralizing Sera on Factor X-Mediated Adenovirus Serotype 5 Gene Transfer","volume":"83","author":"Parker","year":"2009","journal-title":"J. Virol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"4654","DOI":"10.1128\/JVI.02696-06","article-title":"Comparative Seroprevalence and Immunogenicity of Six Rare Serotype Recombinant Adenovirus Vaccine Vectors from Subgroups B and D","volume":"81","author":"Abbink","year":"2007","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2719","DOI":"10.1007\/s13346-023-01362-3","article-title":"Technological Advances in the Use of Viral and Non-Viral Vectors for Delivering Genetic and Non-Genetic Cargos for Cancer Therapy","volume":"13","author":"Dogbey","year":"2023","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1038\/s41571-024-00883-1","article-title":"Nucleic Acid-Based Drugs for Patients with Solid Tumours","volume":"21","author":"Huayamares","year":"2024","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1056\/NEJMoa2035389","article-title":"Efficacy and Safety of the MRNA-1273 SARS-CoV-2 Vaccine","volume":"384","author":"Baden","year":"2021","journal-title":"N. Engl. J. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2603","DOI":"10.1056\/NEJMoa2034577","article-title":"Safety and Efficacy of the BNT162b2 MRNA Covid-19 Vaccine","volume":"383","author":"Polack","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1657","DOI":"10.15585\/mmwr.mm695152e2","article-title":"The Advisory Committee on Immunization Practices\u2019 Updated Interim Recommendation for Allocation of COVID-19 Vaccine-United States, December 2020","volume":"69","author":"Dooling","year":"2021","journal-title":"MMWR Morb. Mortal. Wkly. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Hirai, T., Sato, A., Koizumi, N., Kurioka, Y., Suzuki, Y., Kano, J., Yamakawa, M., Nomura, T., Fujii, M., and Sakurai, F. (2021). The Infectivity of Progeny Adenovirus in the Presence of Neutralizing Antibody. J. Gen. Virol., 102.","DOI":"10.1099\/jgv.0.001590"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1007\/s00430-019-00645-2","article-title":"Human Adenovirus Binding to Host Cell Receptors: A Structural View","volume":"209","author":"Stasiak","year":"2020","journal-title":"Med. Microbiol. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1248\/bpbreports.2.6_113","article-title":"Adenovirus Fiber Can Distribute Itself to the Cell Surface without Membrane Damage","volume":"2","author":"Sato","year":"2019","journal-title":"BPB Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1568","DOI":"10.1126\/science.286.5444.1568","article-title":"Identification of a Conserved Receptor-Binding Site on the Fiber Proteins of CAR-Recognizing Adenoviridae","volume":"286","author":"Roelvink","year":"1999","journal-title":"Science"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1320","DOI":"10.1126\/science.275.5304.1320","article-title":"Isolation of a Common Receptor for Coxsackie B Viruses and Adenoviruses 2 and 5","volume":"275","author":"Bergelson","year":"1997","journal-title":"Science"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1089\/104303403765255165","article-title":"Adenovirus Serotype 5 Fiber Shaft Influences In Vivo Gene Transfer in Mice","volume":"14","author":"Smith","year":"2003","journal-title":"Hum. Gene Ther."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.1089\/104303403322542248","article-title":"Receptor Interactions Involved in Adenoviral-Mediated Gene Delivery After Systemic Administration in Non-Human Primates","volume":"14","author":"Smith","year":"2003","journal-title":"Hum. Gene Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/0092-8674(93)90231-E","article-title":"Integrins Alpha v Beta 3 and Alpha v Beta 5 Promote Adenovirus Internalization but Not Virus Attachment","volume":"73","author":"Wickham","year":"1993","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1089\/hum.2006.17.264","article-title":"Modified Adenoviral Vectors Ablated for Coxsackievirus-Adenovirus Receptor, Alphav Integrin, and Heparan Sulfate Binding Reduce in Vivo Tissue Transduction and Toxicity","volume":"17","author":"Koizumi","year":"2006","journal-title":"Hum. Gene Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"13062","DOI":"10.1128\/JVI.77.24.13062-13072.2003","article-title":"Reduction of Natural Adenovirus Tropism to Mouse Liverby Fiber-Shaft Exchange in Combination with Both CAR-And\u03b1v Integrin-BindingAblation","volume":"77","author":"Koizumi","year":"2003","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Watanabe-Takahashi, M., Tamada, M., Senda, M., Hibino, M., Shimizu, E., Okuta, A., Miyazawa, A., Senda, T., and Nishikawa, K. (2021). Identification of a Peptide Motif That Potently Inhibits Two Functionally Distinct Subunits of Shiga Toxin. Commun. Biol., 4.","DOI":"10.1038\/s42003-021-02068-3"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.jconrel.2011.06.010","article-title":"Design and Cellular Internalization of Genetically Engineered Polypeptide Nanoparticles Displaying Adenovirus Knob Domain","volume":"155","author":"Sun","year":"2011","journal-title":"J. Control. Release"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1159\/000151532","article-title":"Expression and Purification of a Recombinant Adenovirus Fiber Knob in a Baculovirus System","volume":"51","author":"Porchia","year":"2008","journal-title":"Intervirology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"13081","DOI":"10.1073\/pnas.1401437111","article-title":"Nonlytic Viral Spread Enhanced by Autophagy Components","volume":"111","author":"Bird","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/bs.aivir.2020.09.002","article-title":"Viral Cell-to-Cell Spread: Conventional and Non-Conventional Ways","volume":"108","author":"Dutch","year":"2020","journal-title":"Adv. Virus Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1083\/jcb.200503059","article-title":"Actin- and Myosin-Driven Movement of Viruses along Filopodia Precedes Their Entry into Cells","volume":"170","author":"Lehmann","year":"2005","journal-title":"J. Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1038\/ncb1544","article-title":"Retroviruses Can Establish Filopodial Bridges for Efficient Cell-to-Cell Transmission","volume":"9","author":"Sherer","year":"2007","journal-title":"Nat. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Robinson, S.M., Tsueng, G., Sin, J., Mangale, V., Rahawi, S., McIntyre, L.L., Williams, W., Kha, N., Cruz, C., and Hancock, B.M. (2014). Coxsackievirus B Exits the Host Cell in Shed Microvesicles Displaying Autophagosomal Markers. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004045"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"15191","DOI":"10.1073\/pnas.261452898","article-title":"The Coxsackievirus and Adenovirus Receptor Is a Transmembrane Component of the Tight Junction","volume":"98","author":"Cohen","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.jconrel.2016.05.010","article-title":"Exploring the Relationship between Anti-PEG IgM Behaviors and PEGylated Nanoparticles and Its Significance for Accelerated Blood Clearance","volume":"234","author":"Shiraishi","year":"2016","journal-title":"J. Control. Release"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"21560","DOI":"10.1038\/s41598-022-26030-3","article-title":"Effects of Pre-Existing Anti-Adenovirus Antibodies on Transgene Expression Levels and Therapeutic Efficacies of Arming Oncolytic Adenovirus","volume":"12","author":"Ono","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"12315","DOI":"10.1038\/s41598-018-30947-z","article-title":"Antibodies against Adenovirus Fiber and Penton Base Proteins Inhibit Adenovirus Vector-Mediated Transduction in the Liver Following Systemic Administration","volume":"8","author":"Tomita","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1007\/s10735-006-9055-4","article-title":"Coxsackievirus and Adenovirus Receptor Expression in Non-Malignant Lung Tissues and Clinical Lung Cancers","volume":"37","author":"Wang","year":"2006","journal-title":"J. Mol. Histol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1002\/jgm.348","article-title":"Generation of Fiber-Modified Adenovirus Vectors Containing Heterologous Peptides in Both the HI Loop and C Terminus of the Fiber Knob","volume":"5","author":"Koizumi","year":"2003","journal-title":"J. Gene Med."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"730","DOI":"10.1038\/sj.gt.3301453","article-title":"A Simplified System for Constructing Recombinant Adenoviral Vectors Containing Heterologous Peptides in the HI Loop of Their Fiber Knob","volume":"8","author":"Mizuguchi","year":"2001","journal-title":"Gene Ther."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"8221","DOI":"10.1128\/jvi.71.11.8221-8229.1997","article-title":"Increased in Vitro and in Vivo Gene Transfer by Adenovirus Vectors Containing Chimeric Fiber Proteins","volume":"71","author":"Wickham","year":"1997","journal-title":"J. Virol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"7107","DOI":"10.1128\/JVI.75.15.7107-7113.2001","article-title":"Phage Display of Adenovirus Type 5 Fiber Knob as a Tool for Specific Ligand Selection and Validation","volume":"75","author":"Pereboev","year":"2001","journal-title":"J. Virol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1534","DOI":"10.1182\/blood-2009-04-215855","article-title":"A Highly Efficient Short Hairpin RNA Potently Down-Regulates CCR5 Expression in Systemic Lymphoid Organs in the Hu-BLT Mouse Model","volume":"115","author":"Shimizu","year":"2010","journal-title":"Blood"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2577","DOI":"10.1089\/hum.1998.9.17-2577","article-title":"Efficient Construction of a Recombinant Adenovirus Vector by an Improved in Vitro Ligation Method","volume":"9","author":"Mizuguchi","year":"1998","journal-title":"Hum. Gene Ther."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/S0168-3659(02)00059-7","article-title":"Strength Evaluation of Transcriptional Regulatory Elements for Transgene Expression by Adenovirus Vector","volume":"81","author":"Xu","year":"2002","journal-title":"J. Control. Release"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/19\/10679\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:10:09Z","timestamp":1760112609000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/19\/10679"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,3]]},"references-count":46,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2024,10]]}},"alternative-id":["ijms251910679"],"URL":"https:\/\/doi.org\/10.3390\/ijms251910679","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2024,10,3]]}}}