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Distinct Classes of Proteasome-Modulating Agents Cooperatively Augment Recombinant Adeno-Associated Virus Type 2 and Type 5-Mediated Transduction from the Apical Surfaces of Human Airway Epithelia
Open accessAbstract
Tripeptidyl aldehyde proteasome inhibitors have been shown to effectively increase viral capsid ubiquitination and transduction of recombinant adeno-associated virus type 2 (rAAV-2) and rAAV-5 serotypes. In the present study we have characterized a second class of proteasome-modulating agents (anthracycline derivatives) for their ability to induce rAAV transduction. The anthracycline derivatives doxorubicin and aclarubicin were chosen for analysis because they have been shown to interact with the proteasome through a mechanism distinct from that of tripeptidyl aldehydes. Our studies demonstrated that doxorubicin and aclarubicin also significantly augmented rAAV transduction in airway cell lines, polarized human airway epithelia, and mouse lungs. Both tripeptidyl aldehyde and anthracycline proteasome-modulating agents similarly augmented nuclear accumulation of rAAV in A549 and IB3 airway cell lines. However, these two cell types demonstrated cell specificity in the ability of N -acetyl- l -leucyl- l -leucyl- l -norleucine (LLnL) or doxorubicin to augment rAAV transduction. Interestingly, the combined administration of LLnL and doxorubicin resulted in substantially increased transduction (>2,000-fold) following apical infection of human polarized epithelia with either rAAV-2 or rAAV-5. In summary, the cell type specificity of LLnL and doxorubicin to induce rAAV transduction, together with the ability of these compounds to synergistically enhance rAAV transduction in polarized airway epithelial induction, suggests that these two classes of compounds likely modulate different proteasome functions that affect rAAV transduction. Findings from this study provide new insights into how modulation of proteasome function can be effectively used to augment rAAV transduction in airway epithelia for gene therapy of cystic fibrosis.
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“Use of proteasome inhibitors such as MG-132 and LLnL have been shown to increase transduction, providing evidence that after endosome escape, AAV must elude proteasomes in the cytoplasm while traversing to the nucleus (Fig. 8IV and V) (11, 51).”
From Packaging Capacity of Adeno-Associated Virus Serotypes: Impact of Larger Genomes on Infectivity and Postentry Steps (2005) · cites this paper as [51]
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“Studies have been carried out by a number of groups (11, 22, 50, 51) illustrating that drugs/chemicals such as bafilomycin A, brefeldin A, and MG-132 augment rAAV transduction by acting on the level of endosome acidification, early-to-late endosome transition, and proteasome activity, respectively.”
From Packaging Capacity of Adeno-Associated Virus Serotypes: Impact of Larger Genomes on Infectivity and Postentry Steps (2005) · cites this paper as [51]
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“Proteasome inhibitors have been described in numerous studies to increase the transduction of rAAV postentry of the cell, suggesting that proteasomes are a barrier for intracellular trafficking (11, 51).”
From Packaging Capacity of Adeno-Associated Virus Serotypes: Impact of Larger Genomes on Infectivity and Postentry Steps (2005) · cites this paper as [51]
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“Proteasome inhibitors have been described in numerous studies to increase the transduction of rAAV postentry of the cell, suggesting that proteasomes are a barrier for intracellular trafficking (11, 51).”
From Packaging Capacity of Adeno-Associated Virus Serotypes: Impact of Larger Genomes on Infectivity and Postentry Steps (2005) · cites this paper as [51]
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Cites
McCarty, D., Monahan, P., & Samulski, R. (2001) · Gene Therapy · doi:10.1038/sj.gt.3301514 · Cited by 735
Infectious entry pathway of adeno-associated virus and adeno-associated virus vectors
Bartlett, J. S., Wilcher, R., & Samulski, R. J. (2000) · Journal of Virology · doi:10.1128/jvi.74.6.2777-2785.2000 · Cited by 532 · cited 1×
“Studies evaluating rAAV trafficking with the fluorescent and radiolabeled virus have demonstrated that once endocytosed, the virus moves relatively slowly to the nucleus and accumulates in a perinuclear compartment (4, 14, 19, 20, 37).”
BackgroundEndosomal processing limits gene transfer to polarized airway epithelia by adeno-associated virus
Duan, D., Yue, Y., Yan, Z., Yang, J., & Engelhardt, J. F. (2000) · Journal of Clinical Investigation · doi:10.1172/jci8317 · Cited by 391
Self-citationYan, Z., Zak, R., Luxton, G. W. G., Ritchie, T. C., Bantel-Schaal, U., & Engelhardt, J. F. (2002) · Journal of Virology · doi:10.1128/jvi.76.5.2043-2053.2002 · Cited by 222
Self-citationHansen, J., Qing, K., & Srivastava, A. (2001) · Journal of Virology · doi:10.1128/jvi.75.9.4080-4090.2001 · Cited by 129 · cited 1×
“Studies evaluating rAAV trafficking with the fluorescent and radiolabeled virus have demonstrated that once endocytosed, the virus moves relatively slowly to the nucleus and accumulates in a perinuclear compartment (4, 14, 19, 20, 37).”
Background
Research works printed but not linked: 34
- 10.1089/104303401753153956 · DOI
- 10.1172/JCI0215780 · DOI
- 10.1128/JVI.73.7.6085-6088.1999 · DOI
- Berns K. I. and C. Giraud. 1996. Adeno-associated virus (AAV) vectors in gene therapy. Springer Berlin Germany. · DOI
- 10.1023/A:1008323200102 · DOI
- Carter B. J. H. B. Burnstein and R. W. Peluso. AAV vectors for gene therapy in press. Marcel Dekker Inc. New York N.Y.
- Casper, E. S., R. J. Gralla, and C. W. Young. 1981. Clinical phase I study of aclacinomycin A by evaluation of an intermittent intravenous administration schedule. Cancer Res.41:2417-2420.
- 10.1073/pnas.97.7.3428 · DOI · Paper page
- 10.1128/JVI.77.13.7361-7366.2003 · DOI
- Dong, J., M. Naito, T. Tatsuta, H. Seimiya, O. Johdo, and T. Tsuruo. 1995. Difference between the resistance mechanisms of aclacinomycin- and adriamycin-resistant P388 cell lines. Oncol. Res.7:245-252.
- 10.1089/hum.1998.9.18-2761 · DOI
- Duan, D., Y. Zhang, and J. F. Engelhardt. 1998. Gene delivery to the airway, p. 13.9.1-13.9.34. In N. C. Dracopoli, J. L. Haines, B. R. Korf, D. T. Moir, C. C. Morton, C. E. Seidman, J. G. Seidman, and D. R. Smith (ed.), Current protocols in human genetics, John Wiley & Sons, Inc., New York, N.Y.
- 10.1074/jbc.271.28.16455 · DOI
- 10.2165/00003088-200342050-00002 · DOI
- 10.1073/pnas.182412299 · DOI · Paper page
- 10.1128/JVI.74.2.992-996.2000 · DOI
- 10.1089/104303402320139014 · DOI
- 10.1016/0163-7258(90)90088-J · DOI
- 10.1074/jbc.274.6.3363 · DOI
- Kiyomiya, K., M. Kurebe, H. Nakagawa, and S. Matsuo. 2002. The role of the proteasome in apoptosis induced by anthracycline anticancer agents. Int. J. Oncol.20:1205-1209.
- 10.1016/S0024-3205(98)00151-9 · DOI
- 10.1016/S0962-8924(98)01346-4 · DOI · Paper page
- 10.1074/jbc.272.47.29899 · DOI
- Muller, I., D. Niethammer, and G. Bruchelt. 1998. Anthracycline-derived chemotherapeutics in apoptosis and free radical cytotoxicity. Int. J. Mol. Med.1:491-494.
- 10.1128/JVI.76.24.12634-12645.2002 · DOI
- 10.1056/NEJM199809243391307 · DOI
- 10.1053/ctrv.2002.0262 · DOI
- 10.1021/bi00154a014 · DOI
- 10.1097/00005537-199902000-00017 · DOI
- 10.1089/hum.1998.9.6-889 · DOI
- 10.1089/104303402760128577 · DOI
- 10.1128/JVI.76.22.11505-11517.2002 · DOI
- 10.1016/S0076-6879(02)46065-X · DOI
- 10.1128/JVI.74.8.3852-3858.2000 · DOI
Cited by
Sorted by citations
Items 101–122 of 122
PEG-mediated transduction of rAAV as a platform for spatially confined and efficient gene delivery
Zou, L., Wang, J., Fāng, Y., & Tian, H. (2022) · Biomaterials Research · doi:10.1186/s40824-022-00322-1 · Cited by 7
“In previous studies, a series of compounds have been used as rAAV transduction enhancer, including topoisomerase and proteasome inhibitors, epipodophyllotoxins, inducers of DNA damage, effectors of epigenetic modification, and anthracyclines [26, 40, 41].”
BackgroundEffect of cytoskeleton inhibitors on deadhesion kinetics of HepG2 cells on biomimetic surface.
Feng, Z.-Q., Chen, B., Tang, S.-C., Liao, K., Chen, W.-N., & Chan, V. (2010) · Colloids and Surfaces B: Biointerfaces · doi:10.1016/j.colsurfb.2009.08.010 · Cited by 7
Ning, K., Zhang, X. J., Feng, Z., Hao, S., Küz, Ç. A., Cheng, F., Park, S. Y., McFarlin, S., Engelhardt, J. F., Yan, Z., et al. (2023) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2023.101115 · Cited by 8
“41 In this study, we investigated the DDR activation following rAAV2.”
BackgroundMethodInfluentialSelf-citationRecent Advancements in AAV-Vectored Immunoprophylaxis in the Nonhuman Primate Model
Campbell, E. S. B., Goens, M. M., Cao, W., Thompson, B., Susta, L., Banadyga, L., & Wootton, S. K. (2023) · Biomedicines · doi:10.3390/biomedicines11082223 · Cited by 9
Cystic fibrosis: potential options for gene-directed therapies
Moraes, T., & Downey, G. (2004) · Drug Discovery Today Therapeutic Strategies · doi:10.1016/j.ddstr.2004.11.009 · Cited by 4
Targeted Gene Insertion for Functional CFTR Restoration in Airway Epithelium
Barillà, C., Suzuki, S., Rab, A., Sorscher, E. J., & Davis, B. R. (2022) · Frontiers in Genome Editing · doi:10.3389/fgeed.2022.847645 · Cited by 6
“Treatment of human airway epithelium by transiently disrupting tight junctions or inhibiting proteasomal processing facilitates AAV transduction (Coyne et al., 2000; Duan et al., 2000; Yan et al., 2004; Ellis et al., 2013).”
BackgroundLong Period Modeling SARS-CoV-2 Infection of in Vitro Cultured Polarized Human Airway Epithelium
Hao, S.-Y., Ning, K., Kuz, C. A., Vorhies, K., Yan, Z.-Y., & Qiu, J.-M. (2020) · bioRxiv · doi:10.1101/2020.08.27.271130 · Cited by 3
Self-citationPreclinical Study Design for rAAV
Flotte, T. R., Conlon, T. J., & Mueller, C. (2011) · Methods in molecular biology · doi:10.1007/978-1-61779-370-7_14 · Cited by 3
Mechanisms of AAV neutralization by human alpha-defensins
Porter, J. M., Hulce, K. R., Oswald, M. S., Busuttil, K., Emmanuel, S. N., Bennett, A. D., McKenna, R., & Smith, J. G. (2025) · PLoS Pathogens · doi:10.1371/journal.ppat.1013283 · Cited by 2
“These mechanisms are not mutually exclusive, although we note that all samples are treated with the proteosome inhibitor doxorubicin, which has been shown to increase AAV transduction efficiency [90].”
Bleomycin promotes rAAV2 transduction via DNA-PKcs/Artemis-mediated DNA break repair pathways
Huang, X., Wang, X., Sun, Y., Li, L., Li, A., Xu, W., Xie, X., & Diao, Y. (2023) · Virology · doi:10.1016/j.virol.2023.109959 · Cited by 2
Rajapaksha, I. G., Angus, P., & Herath, C. B. (2019) · IntechOpen eBooks · doi:10.5772/intechopen.80317 · Cited by 5
“Also, the application of different classes of proteasome inhibitors such as tripeptidyl aldehydes and N-acetyl-l-leucyl-l-leucyl-l-norleucinal (LLnL) and the anthracycline compounds such as doxorubicin increases the rate of viral translocation to the nucleus [32].”
BackgroundA systemically delivered AAV-CFTR gene therapy for cystic fibrosisPreprint
Plasschaert, L. W., Stutz, C., Otarola, E., Ruggeri, L., Misiolek, R. V., Nuccio, A. G., Shang, J., Choo-Wing, R., Taddese, B., Decock, A., et al. (2025) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2025.03.20.642115
Hao, S.-Y., Zhang, X., Ning, K., Feng, Z., Park, S. Y., Kuz, C. A., McFarlin, S., Richart, D., Cheng, F., Zhang, E. Y., et al. (2023) · bioRxiv · doi:10.1101/2023.09.27.559795 · Cited by 1
“288 Doxorubicin (Dox), an FDA approved drug for cancer therapy, has been used as a 289 potent agent to augment AAV-mediated transgene expression, especially in airway epithelia, 290 functioning as a proteasome inhibitor to prevent the degradation of ubiquitinated AAV in the 291 cytoplasm and facilitate vector nuclear…”
MethodResultSelf-citationKienle, E. (2014) · heiDOK (Heidelberg University) · doi:10.11588/heidok.00017851 · Cited by 1
“To some extent, AAV capsids are subject to ubiquitination and degradation, as evidenced by findings that inhibition of the proteasome enhances cell transduction with several AAV serotypes [121], [129]–[134].”
BackgroundMethodpH-responsive virus-drug conjugates for targeted PD-L1 silencing and enhanced cancer immunotherapy
Tseng, S.-J., Kempson, I., Ho, Y., Yang, P.-C., & Liao, Z.-X. (2026) · Journal of Nanobiotechnology · doi:10.1186/s12951-026-04347-9
“Consistent with the action of other DNA-damaging agents [27, 28], Dox primarily promotes the formation of stable, transcription-ready dsDNA episomes from the viral genome [6, 29].”
Modulation of AAV transduction and integration targeting by topoisomerase poisons
Kasimsetty, A., Hwang, Y., Everett, J. K., McFarland, A. G., Zolnoski, S. A., Lu, T., Roche, A. M., Martínez‐García, P. M., Sabatino, D. E., & Bushman, F. D. (2024) · Molecular Therapy — Methods & Clinical Development · doi:10.1016/j.omtm.2024.101364 · Cited by 2
“15,16 DOX was reported to enhance AAV transduction 17 in several different tissues and animal models.”
Gong, H., Yuan, N., Shen, Z., Tang, C., Shipp, S. D., Qian, L., Lu, Y., Andolina, I. M., Zhang, S., Wu, J., et al. (2020) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2020.06.08.139667 · Cited by 1
“Furthermore, doxorubicin may also promote viral nuclear entry through modulation of proteasome function (Yan et al., 2004).”
BackgroundInfluentialYu, C.-Y., Wang, B., Wang, Z., Cao, Z., Li, J., & Xiao, X. (2006) · Molecular Therapy · doi:10.1016/j.ymthe.2006.08.556
Strategies for improving adeno-associated viral infection of airway epithelial cells
Dickey, D. D. (2012) · doi:10.17077/etd.m1aiqczb
“studies that utilized proteasomal inhibitors to augment AAV transduction (143).”
Identification of ebola glycoprotein mutants that exhibit increased transduction efficiency
Sandersfeld, L. M. (2009) · doi:10.17077/etd.7awu17s3
“Finally, AAV vectors have been shown to be degraded by bronchial secretions in injured or diseased lungs of CF patients, mostly due to proteasome degradation (96, 103, 107).”
Background
Metadata
- Authors
- Yan, Z., Zak, R., Zhang, Y., Ding, W., Godwin, S., Munson, K., Peluso, R., & Engelhardt, J. F.
- Affiliations
- University of Iowa (US)
- Department of Anatomy and Cell Biology
- Targeted Genetics Corporation, Seattle, Washington 98101
- Corresponding author
- Engelhardt, John F.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2004-03-15
- Volume
- 78
- Issue
- 6
- Pages
- 2863-2874
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC353734
- OpenAlex
- W1982346550
- Cited by
- 138
- References
- 0
- Citations by year
- 2026: 3 · 2025: 2 · 2024: 7 · 2023: 7 · 2022: 4 · 2021: 3 · 2020: 8 · 2019: 7 · 2018: 6 · 2017: 1 · 2016: 5 · 2015: 3 · 2014: 4 · 2013: 5 · 2012: 6
- FWCI
- 6.78
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of Virology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Heart, Lung, and Blood Institute (R01HL058340, R01 HL58340)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK54759, P30DK054759)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Keywords
- proteasome inhibitors, adeno-associated virus type 2, doxorubicin, aclarubicin, Lawrence Livermore National Laboratory
- MeSH terms
- Dependovirus, Animals, Cell Line, Cysteine Endopeptidases, Cystic Fibrosis, Doxorubicin, Epithelial Cells, Humans, Leupeptins, Lung, Mice, Inbred C57BL, Multienzyme Complexes, Parvoviridae Infections, Recombination, Genetic, Transduction, Genetic, Aclarubicin, Genetic Therapy, Cysteine Proteinase Inhibitors, Cell Polarity, Proteasome Endopeptidase Complex, Mice
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