Adenovirus-Neutralizing and Infection-Promoting Activities Measured in Serum of Human Brain Cancer Patients Treated with Oncolytic Adenovirus Ad5-∆24.RGD
Abstract
1. Introduction
2. Results
2.1. Neutralizing Antibody Titers in Patients with Glioblastoma Undergoing Treatment with Oncolytic Adenovirus Ad5-∆24.RGD
2.2. Detection of a Blood Component That Promotes RGD-Mediated Adenovirus Infection
2.3. Partial Characterization of the Blood Component That Promotes RGD-Mediated Adenovirus Infection
3. Discussion
4. Materials and Methods
4.1. Collection and Preparation of Human Serum, Plasma and Cerebrospinal Fluid Samples
4.2. Adenovirus Neutralization Assay
4.3. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Farrera-Sal, M.; Moya-Borrego, L.; Bazan-Peregrino, M.; Alemany, R. Evolving Status of Clinical Immunotherapy with Oncolytic Adenovirus. Clin. Cancer Res. 2021, 27, 2979–2988. [Google Scholar] [CrossRef] [PubMed]
- Mantwill, K.; Klein, F.G.; Wang, D.; Hindupur, S.V.; Ehrenfeld, M.; Holm, P.S.; Nawroth, R. Concepts in Oncolytic Adenovirus Therapy. Int. J. Mol. Sci. 2021, 22, 10522. [Google Scholar] [CrossRef]
- Qiu, Q.; Xu, Z.; Tian, J.; Moitra, R.; Gunti, S.; Notkins, A.L.; Byrnes, A.P. Impact of natural IgM concentration on gene therapy with adenovirus type 5 vectors. J. Virol. 2015, 89, 3412–3416. [Google Scholar] [CrossRef]
- Xu, Z.; Tian, J.; Smith, J.S.; Byrnes, A.P. Clearance of adenovirus by Kupffer cells is mediated by scavenger receptors, natural antibodies, and complement. J. Virol. 2008, 82, 11705–11713. [Google Scholar] [CrossRef]
- Vogels, R.; Zuijdgeest, D.; van Rijnsoever, R.; Hartkoorn, E.; Damen, I.; de Bethune, M.P.; Kostense, S.; Penders, G.; Helmus, N.; Koudstaal, W.; et al. Replication-deficient human adenovirus type 35 vectors for gene transfer and vaccination: Efficient human cell infection and bypass of preexisting adenovirus immunity. J. Virol. 2003, 77, 8263–8271. [Google Scholar] [CrossRef] [PubMed]
- Tsai, V.; Johnson, D.E.; Rahman, A.; Wen, S.F.; LaFace, D.; Philopena, J.; Nery, J.; Zepeda, M.; Maneval, D.C.; Demers, G.W.; et al. Impact of human neutralizing antibodies on antitumor efficacy of an oncolytic adenovirus in a murine model. Clin. Cancer Res. 2004, 10, 7199–7206. [Google Scholar] [CrossRef]
- Ono, R.; Nishimae, F.; Wakida, T.; Sakurai, F.; Mizuguchi, H. Effects of pre-existing anti-adenovirus antibodies on transgene expression levels and therapeutic efficacies of arming oncolytic adenovirus. Sci. Rep. 2022, 12, 21560. [Google Scholar] [CrossRef]
- Holterman, L.; Vogels, R.; van der Vlugt, R.; Sieuwerts, M.; Grimbergen, J.; Kaspers, J.; Geelen, E.; van der Helm, E.; Lemckert, A.; Gillissen, G.; et al. Novel replication-incompetent vector derived from adenovirus type 11 (Ad11) for vaccination and gene therapy: Low seroprevalence and non-cross-reactivity with Ad5. J. Virol. 2004, 78, 13207–13215. [Google Scholar] [CrossRef] [PubMed]
- Roy, S.; Shirley, P.S.; McClelland, A.; Kaleko, M. Circumvention of immunity to the adenovirus major coat protein hexon. J. Virol. 1998, 72, 6875–6879. [Google Scholar] [CrossRef] [PubMed]
- Wu, H.; Dmitriev, I.; Kashentseva, E.; Seki, T.; Wang, M.; Curiel, D.T. Construction and characterization of adenovirus serotype 5 packaged by serotype 3 hexon. J. Virol. 2002, 76, 12775–12782. [Google Scholar] [CrossRef]
- Bots, S.T.F.; Kemp, V.; Cramer, S.J.; van den Wollenberg, D.J.M.; Hornsveld, M.; Lamfers, M.L.M.; van der Pluijm, G.; Hoeben, R.C. Nonhuman Primate Adenoviruses of the Human Adenovirus B Species Are Potent and Broadly Acting Oncolytic Vector Candidates. Hum. Gene Ther. 2022, 33, 275–289. [Google Scholar] [CrossRef] [PubMed]
- Cheng, T.; Song, Y.; Zhang, Y.; Zhang, C.; Yin, J.; Chi, Y.; Zhou, D. A novel oncolytic adenovirus based on simian adenovirus serotype 24. Oncotarget 2017, 8, 26871–26885. [Google Scholar] [CrossRef]
- Hemminki, A.; Kanerva, A.; Kremer, E.J.; Bauerschmitz, G.J.; Smith, B.F.; Liu, B.; Wang, M.; Desmond, R.A.; Keriel, A.; Barnett, B.; et al. A canine conditionally replicating adenovirus for evaluating oncolytic virotherapy in a syngeneic animal model. Mol. Ther. 2003, 7, 163–173. [Google Scholar] [CrossRef]
- Chen, J.; Gao, P.; Yuan, S.; Li, R.; Ni, A.; Chu, L.; Ding, L.; Sun, Y.; Liu, X.Y.; Duan, Y. Oncolytic Adenovirus Complexes Coated with Lipids and Calcium Phosphate for Cancer Gene Therapy. ACS Nano 2016, 10, 11548–11560. [Google Scholar] [CrossRef] [PubMed]
- Croyle, M.A.; Chirmule, N.; Zhang, Y.; Wilson, J.M. “Stealth” adenoviruses blunt cell-mediated and humoral immune responses against the virus and allow for significant gene expression upon readministration in the lung. J. Virol. 2001, 75, 4792–4801. [Google Scholar] [CrossRef]
- Rojas, L.A.; Condezo, G.N.; Moreno, R.; Fajardo, C.A.; Arias-Badia, M.; San Martin, C.; Alemany, R. Albumin-binding adenoviruses circumvent pre-existing neutralizing antibodies upon systemic delivery. J. Control Release 2016, 237, 78–88. [Google Scholar] [CrossRef]
- Lv, P.; Liu, X.; Chen, X.; Liu, C.; Zhang, Y.; Chu, C.; Wang, J.; Wang, X.; Chen, X.; Liu, G. Genetically Engineered Cell Membrane Nanovesicles for Oncolytic Adenovirus Delivery: A Versatile Platform for Cancer Virotherapy. Nano Lett. 2019, 19, 2993–3001. [Google Scholar] [CrossRef] [PubMed]
- Gahery-Segard, H.; Farace, F.; Godfrin, D.; Gaston, J.; Lengagne, R.; Tursz, T.; Boulanger, P.; Guillet, J.G. Immune response to recombinant capsid proteins of adenovirus in humans: Antifiber and anti-penton base antibodies have a synergistic effect on neutralizing activity. J. Virol. 1998, 72, 2388–2397. [Google Scholar] [CrossRef]
- Hong, S.S.; Habib, N.A.; Franqueville, L.; Jensen, S.; Boulanger, P.A. Identification of adenovirus (ad) penton base neutralizing epitopes by use of sera from patients who had received conditionally replicative ad (addl1520) for treatment of liver tumors. J. Virol. 2003, 77, 10366–10375. [Google Scholar] [CrossRef]
- Sumida, S.M.; Truitt, D.M.; Lemckert, A.A.; Vogels, R.; Custers, J.H.; Addo, M.M.; Lockman, S.; Peter, T.; Peyerl, F.W.; Kishko, M.G.; et al. Neutralizing antibodies to adenovirus serotype 5 vaccine vectors are directed primarily against the adenovirus hexon protein. J. Immunol. 2005, 174, 7179–7185. [Google Scholar] [CrossRef] [PubMed]
- Koski, A.; Kangasniemi, L.; Escutenaire, S.; Pesonen, S.; Cerullo, V.; Diaconu, I.; Nokisalmi, P.; Raki, M.; Rajecki, M.; Guse, K.; et al. Treatment of cancer patients with a serotype 5/3 chimeric oncolytic adenovirus expressing GMCSF. Mol. Ther. 2010, 18, 1874–1884. [Google Scholar] [CrossRef] [PubMed]
- Kimball, K.J.; Preuss, M.A.; Barnes, M.N.; Wang, M.; Siegal, G.P.; Wan, W.; Kuo, H.; Saddekni, S.; Stockard, C.R.; Grizzle, W.E.; et al. A phase I study of a tropism-modified conditionally replicative adenovirus for recurrent malignant gynecologic diseases. Clin. Cancer Res. 2010, 16, 5277–5287. [Google Scholar] [CrossRef]
- Lang, F.F.; Conrad, C.; Gomez-Manzano, C.; Yung, W.K.A.; Sawaya, R.; Weinberg, J.S.; Prabhu, S.S.; Rao, G.; Fuller, G.N.; Aldape, K.D.; et al. Phase I Study of DNX-2401 (Delta-24-RGD) Oncolytic Adenovirus: Replication and Immunotherapeutic Effects in Recurrent Malignant Glioma. J. Clin. Oncol. 2018, 36, 1419–1427. [Google Scholar] [CrossRef] [PubMed]
- van Putten, E.H.P.; Kleijn, A.; van Beusechem, V.W.; Noske, D.; Lamers, C.H.J.; de Goede, A.L.; Idema, S.; Hoefnagel, D.; Kloezeman, J.J.; Fueyo, J.; et al. Convection Enhanced Delivery of the Oncolytic Adenovirus Delta24-RGD in Patients with Recurrent GBM: A Phase I Clinical Trial Including Correlative Studies. Clin. Cancer Res. 2022, 28, 1572–1585. [Google Scholar] [CrossRef] [PubMed]
- Garcia, M.; Moreno, R.; Gil-Martin, M.; Cascallo, M.; de Olza, M.O.; Cuadra, C.; Piulats, J.M.; Navarro, V.; Domenech, M.; Alemany, R.; et al. A Phase 1 Trial of Oncolytic Adenovirus ICOVIR-5 Administered Intravenously to Cutaneous and Uveal Melanoma Patients. Hum. Gene Ther. 2019, 30, 352–364. [Google Scholar] [CrossRef] [PubMed]
- Fares, J.; Ahmed, A.U.; Ulasov, I.V.; Sonabend, A.M.; Miska, J.; Lee-Chang, C.; Balyasnikova, I.V.; Chandler, J.P.; Portnow, J.; Tate, M.C.; et al. Neural stem cell delivery of an oncolytic adenovirus in newly diagnosed malignant glioma: A first-in-human, phase 1, dose-escalation trial. Lancet Oncol. 2021, 22, 1103–1114. [Google Scholar] [CrossRef] [PubMed]
- Wang, M.; Hemminki, A.; Siegal, G.P.; Barnes, M.N.; Dmitriev, I.; Krasnykh, V.; Liu, B.; Curiel, D.T.; Alvarez, R.D. Adenoviruses with an RGD-4C modification of the fiber knob elicit a neutralizing antibody response but continue to allow enhanced gene delivery. Gynecol. Oncol. 2005, 96, 341–348. [Google Scholar] [CrossRef]
- Sarkioja, M.; Pesonen, S.; Raki, M.; Hakkarainen, T.; Salo, J.; Ahonen, M.T.; Kanerva, A.; Hemminki, A. Changing the adenovirus fiber for retaining gene delivery efficacy in the presence of neutralizing antibodies. Gene Ther. 2008, 15, 921–929. [Google Scholar] [CrossRef] [PubMed]
- Raki, M.; Sarkioja, M.; Escutenaire, S.; Kangasniemi, L.; Haavisto, E.; Kanerva, A.; Cerullo, V.; Joensuu, T.; Oksanen, M.; Pesonen, S.; et al. Switching the fiber knob of oncolytic adenoviruses to avoid neutralizing antibodies in human cancer patients. J. Gene Med. 2011, 13, 253–261. [Google Scholar] [CrossRef]
- Ruoslahti, E. RGD and other recognition sequences for integrins. Annu. Rev. Cell Dev. Biol. 1996, 12, 697–715. [Google Scholar] [CrossRef]
- Herz, J.; Gerard, R.D. Adenovirus-mediated transfer of low density lipoprotein receptor gene acutely accelerates cholesterol clearance in normal mice. Proc. Natl. Acad. Sci. USA 1993, 90, 2812–2816. [Google Scholar] [CrossRef] [PubMed]
- Dmitriev, I.; Krasnykh, V.; Miller, C.R.; Wang, M.; Kashentseva, E.; Mikheeva, G.; Belousova, N.; Curiel, D.T. An adenovirus vector with genetically modified fibers demonstrates expanded tropism via utilization of a coxsackievirus and adenovirus receptor-independent cell entry mechanism. J. Virol. 1998, 72, 9706–9713. [Google Scholar] [CrossRef] [PubMed]
- Cichon, G.; Boeckh-Herwig, S.; Schmidt, H.H.; Wehnes, E.; Muller, T.; Pring-Akerblom, P.; Burger, R. Complement activation by recombinant adenoviruses. Gene Ther. 2001, 8, 1794–1800. [Google Scholar] [CrossRef] [PubMed]
- Bottermann, M.; Foss, S.; Caddy, S.L.; Clift, D.; van Tienen, L.M.; Vaysburd, M.; Cruickshank, J.; O’Connell, K.; Clark, J.; Mayes, K.; et al. Complement C4 Prevents Viral Infection through Capsid Inactivation. Cell Host Microbe 2019, 25, 617–629 e617. [Google Scholar] [CrossRef] [PubMed]
- Tam, J.C.; Bidgood, S.R.; McEwan, W.A.; James, L.C. Intracellular sensing of complement C3 activates cell autonomous immunity. Science 2014, 345, 1256070. [Google Scholar] [CrossRef] [PubMed]
- Simon, J.; Muller, J.; Ghazaryan, A.; Morsbach, S.; Mailander, V.; Landfester, K. Protein denaturation caused by heat inactivation detrimentally affects biomolecular corona formation and cellular uptake. Nanoscale 2018, 10, 21096–21105. [Google Scholar] [CrossRef]
- Soltis, R.D.; Hasz, D.; Morris, M.J.; Wilson, I.D. The effect of heat inactivation of serum on aggregation of immunoglobulins. Immunology 1979, 36, 37–45. [Google Scholar]
- Lesniak, A.; Campbell, A.; Monopoli, M.P.; Lynch, I.; Salvati, A.; Dawson, K.A. Serum heat inactivation affects protein corona composition and nanoparticle uptake. Biomaterials 2010, 31, 9511–9518. [Google Scholar] [CrossRef] [PubMed]
- Parker, A.L.; Waddington, S.N.; Nicol, C.G.; Shayakhmetov, D.M.; Buckley, S.M.; Denby, L.; Kemball-Cook, G.; Ni, S.; Lieber, A.; McVey, J.H.; et al. Multiple vitamin K-dependent coagulation zymogens promote adenovirus-mediated gene delivery to hepatocytes. Blood 2006, 108, 2554–2561. [Google Scholar] [CrossRef] [PubMed]
- Shin, D.H.; Jiang, H.; Gillard, A.G.; Kim, D.; Fan, X.; Singh, S.K.; Nguyen, T.T.; Sohoni, S.S.; Lopez-Rivas, A.R.; Parthasarathy, A.; et al. Chimeric oncolytic adenovirus evades neutralizing antibodies from human patients and exhibits enhanced anti-glioma efficacy in immunized mice. Mol. Ther. 2024, 32, 722–733. [Google Scholar] [CrossRef] [PubMed]
- Sprangers, M.C.; Lakhai, W.; Koudstaal, W.; Verhoeven, M.; Koel, B.F.; Vogels, R.; Goudsmit, J.; Havenga, M.J.; Kostense, S. Quantifying adenovirus-neutralizing antibodies by luciferase transgene detection: Addressing preexisting immunity to vaccine and gene therapy vectors. J. Clin. Microbiol. 2003, 41, 5046–5052. [Google Scholar] [CrossRef] [PubMed]
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van der Meulen-Muileman, I.H.; Amado-Azevedo, J.; Lamfers, M.L.M.; Kleijn, A.; Idema, S.; Noske, D.P.; Dirven, C.M.F.; van Beusechem, V.W. Adenovirus-Neutralizing and Infection-Promoting Activities Measured in Serum of Human Brain Cancer Patients Treated with Oncolytic Adenovirus Ad5-∆24.RGD. Int. J. Mol. Sci. 2025, 26, 854. https://doi.org/10.3390/ijms26020854
van der Meulen-Muileman IH, Amado-Azevedo J, Lamfers MLM, Kleijn A, Idema S, Noske DP, Dirven CMF, van Beusechem VW. Adenovirus-Neutralizing and Infection-Promoting Activities Measured in Serum of Human Brain Cancer Patients Treated with Oncolytic Adenovirus Ad5-∆24.RGD. International Journal of Molecular Sciences. 2025; 26(2):854. https://doi.org/10.3390/ijms26020854
Chicago/Turabian Stylevan der Meulen-Muileman, Ida H., Joana Amado-Azevedo, Martine L. M. Lamfers, Anne Kleijn, Sander Idema, David P. Noske, Clemens M. F. Dirven, and Victor W. van Beusechem. 2025. "Adenovirus-Neutralizing and Infection-Promoting Activities Measured in Serum of Human Brain Cancer Patients Treated with Oncolytic Adenovirus Ad5-∆24.RGD" International Journal of Molecular Sciences 26, no. 2: 854. https://doi.org/10.3390/ijms26020854
APA Stylevan der Meulen-Muileman, I. H., Amado-Azevedo, J., Lamfers, M. L. M., Kleijn, A., Idema, S., Noske, D. P., Dirven, C. M. F., & van Beusechem, V. W. (2025). Adenovirus-Neutralizing and Infection-Promoting Activities Measured in Serum of Human Brain Cancer Patients Treated with Oncolytic Adenovirus Ad5-∆24.RGD. International Journal of Molecular Sciences, 26(2), 854. https://doi.org/10.3390/ijms26020854