Liver Transduction with Recombinant Adeno-Associated Virus Is Primarily Restricted by Capsid Serotype Not Vector Genotype
Open accessAbstract
We and others have recently reported highly efficient liver gene transfer with adeno-associated virus 8 (AAV-8) pseudotypes, i.e., AAV-2 genomes packaged into AAV-8 capsids. Here we studied whether liver transduction could be further enhanced by using viral DNA packaging sequences (inverted terminal repeats [ITRs]) derived from AAV genotypes other than 2. To this end, we generated two sets of vector constructs carrying expression cassettes embedding a gfp gene or the human factor IX ( hfIX ) gene flanked by ITRs from AAV genotypes 1 through 6. Initial in vitro analyses of gfp vector DNA replication, encapsidation, and cell transduction revealed a surprisingly high degree of interchangeability among the six genotypes. For subsequent in vivo studies, we cross-packaged the six hfIX variants into AAV-8 and infused mice via the portal vein with doses of 5 × 10 10 to 1.8 × 10 12 particles. Notably, all vectors expressed comparably high plasma hFIX levels within a dose cohort over the following 6 months, concurrent with the finding of equivalent vector DNA copy numbers per cell. Partial hepatectomies resulted in ∼80% drops of hFIX levels and vector DNA copy numbers in all groups, indicating genotype-independent persistence of predominantly episomal vector DNA. Southern blot analyses of total liver DNA in fact confirmed the presence of identical and mostly nonintegrated molecular vector forms for all genotypes. We conclude that, unlike serotypes, AAV genotypes are not critical for efficient hepatocyte transduction and can be freely substituted. This corroborates our current model for AAV vector persistence in the liver and provides useful information for the future design and application of recombinant AAV.
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AAV2 Rep proteins can replicate and cross-package AAV genomes with ITRs from serotypes 1, 3, 4, and 6 into non-AAV2 capsids.
“Even with these differences, the AAV2 Reps are capable of replicating and cross-packaging genomes from serotypes 1, 3, 4, and 6 into numerous, non-AAV2 capsids.9,10”
From Adeno-Associated Virus Serotype-Specific Inverted Terminal Repeat Sequence Role in Vector Transgene Expression (2020) · cites this paper as [10]
In the context of a strong ubiquitous promoter, ITR sequences would likely have no effect on overall transgene production.
“In the context of a strong ubiquitous promoter, these ITR sequences would likely have no effect on overall transgene production, as previously shown,10”
From Adeno-Associated Virus Serotype-Specific Inverted Terminal Repeat Sequence Role in Vector Transgene Expression (2020) · cites this paper as [10]
Cites
Ferrari, F. K., Samulski, T., Shenk, T., & Samulski, R. J. (1996) · Journal of Virology · doi:10.1128/jvi.70.5.3227-3234.1996 · Cited by 761 · see citing works
Gene therapy vectors based on adeno-associated virus type 1
Xiao, W., Chirmule, N., Berta, S. C., McCullough, B., Gao, G., & Wilson, J. M. (1999) · Journal of Virology · doi:10.1128/jvi.73.5.3994-4003.1999 · Cited by 541 · see citing works
Grimm, D., Kay, M. A., & Kleinschmidt, J. A. (2003) · Molecular Therapy · doi:10.1016/s1525-0016(03)00095-9 · Cited by 344 · see citing works
Self-citationSamulski, R. J., Srivastava, A., Berns, K. I., & Muzyczka, N. (1983) · Cell · doi:10.1016/0092-8674(83)90342-2 · Cited by 297 · see citing works
Adeno-associated virus vectors integrate at chromosome breakage sites
Miller, D. G., Petek, L. M., & Russell, D. W. (2004) · Nature Genetics · doi:10.1038/ng1380 · Cited by 266 · see citing works
Bleker, S., Sonntag, F., & Kleinschmidt, J. A. (2005) · Journal of Virology · doi:10.1128/jvi.79.4.2528-2540.2005 · Cited by 200 · cited 1× · see citing works
“Since the eight plasmids mainly differed in VP1 expression, we speculate that, similar to AAV-2 (2, 22, 31), the phospholipase 2A domain in the VP1 N terminus is crucial for AAV-8 particle infectivity.”
BackgroundGrimm, D., Kern, A., Pawlita, M., Ferrari, F. K., Samulski, R. J., & Kleinschmidt, J. A. (1999) · Gene Therapy · doi:10.1038/sj.gt.3300946 · Cited by 195 · see citing works
Self-citationLarge‐scale molecular characterization of adeno‐associated virus vector integration in mouse liver
Nakai, H., Wu, X., Fuess, S., Storm, T. A., Munroe, D., Montini, E., Burgess, S. M., Grompe, M., & Kay, M. A. (2005) · Journal of Virology · doi:10.1128/jvi.79.6.3606-3614.2005 · Cited by 190 · see citing works
Self-citationWang, X. S., Ponnazhagan, S., & Srivastava, A. (1996) · Journal of Virology · doi:10.1128/jvi.70.3.1668-1677.1996 · Cited by 104 · see citing works
Structural Analysis of Adeno-Associated Virus Transduction Circular Intermediates
Duan, D., Yan, Z., Yue, Y., & Engelhardt, J. F. (1999) · Virology · doi:10.1006/viro.1999.9821 · Cited by 100 · see citing works
Wang, X. S., Qing, K., Ponnazhagan, S., & Srivastava, A. (1997) · Journal of Virology · doi:10.1128/jvi.71.4.3077-3082.1997 · Cited by 61 · see citing works
Nakai, H., Montini, E., Fuess, S., Storm, T. A., Meuse, L., Finegold, M., Grompe, M., & Kay, M. A. (2003) · Molecular Therapy · doi:10.1016/s1525-0016(02)00023-0 · Cited by 52 · see citing works
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Analysis of AAV Serotypes 1–9 Mediated Gene Expression and Tropism in Mice After Systemic Injection
Zincarelli, C., Soltys, S., Rengo, G., & Rabinowitz, J. E. (2008) · Molecular Therapy · doi:10.1038/mt.2008.76 · Cited by 1,490 · see citing works
Grimm, D., Lee, J. S., Wang, L., Desai, T., Akache, B., Storm, T. A., & Kay, M. A. (2008) · Journal of Virology · doi:10.1128/jvi.00254-08 · Cited by 696 · see citing works
Self-citationHuman Immune Responses to Adeno-Associated Virus (AAV) Vectors
Ronzitti, G., Gross, D.-A., & Mingozzi, F. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.00670 · Cited by 357 · see citing works
Basic Biology of Adeno-Associated Virus (AAV) Vectors Used in Gene Therapy
Balakrishnan, B., & Jayandharan, G. (2014) · Current Gene Therapy · doi:10.2174/1566523214666140302193709 · Cited by 225 · see citing works
Qiao, C., Yuan, Z., Li, J., He, B., Zheng, H., Mayer, C., Li, J., & Xiao, X. (2011) · Gene Therapy · doi:10.1038/gt.2010.157 · Cited by 163 · see citing works
Tailoring the AAV vector capsid for gene therapy
Vandenberghe, L. H., Wilson, J. M., & Gao, G. (2009) · Gene Therapy · doi:10.1038/gt.2008.170 · Cited by 155 · see citing works
Next-generation AAV vectors—Do not judge a virus (only) by its cover
Domenger, C., & Grimm, D. (2019) · Human Molecular Genetics · doi:10.1093/hmg/ddz148 · Cited by 147 · see citing works
Self-citationAdeno‐associated virus gene repair corrects a mouse model of hereditary tyrosinemia in vivo†‡
Paulk, N. K., Wursthorn, K., Wang, Z., Finegold, M. J., Kay, M. A., & Grompe, M. (2010) · Hepatology · doi:10.1002/hep.23481 · Cited by 135 · see citing works
Self-citationEarley, L. F., Conatser, L. M., Lue, V. M., Dobbins, A. L., Li, C., Hirsch, M. L., & Samulski, R. J. (2020) · Human Gene Therapy · doi:10.1089/hum.2019.274 · Cited by 116 · see citing works
Pañeda, A., Vanrell, L., Mauleon, I., Crettaz, J. S., Berraondo, P., Timmermans, E. J., Beattie, S. G., Twisk, J., van Deventer, S., Prieto, J., et al. (2009) · Human Gene Therapy · doi:10.1089/hum.2009.031 · Cited by 109 · see citing works
Schnödt, M., & Büning, H. (2017) · Human Gene Therapy Methods · doi:10.1089/hgtb.2016.188 · Cited by 77 · see citing works
Rational engineering of a functional CpG-free ITR for AAV gene therapy
Pan, X., Yue, Y., Boftsi, M., Wasala, L. P., Tran, N. T., Zhang, K., Pintel, D. J., Tai, P. W. L., & Duan, D. (2022) · Gene Therapy · doi:10.1038/s41434-021-00296-0 · Cited by 53 · see citing works
Hu, C., & Lipshutz, G. S. (2012) · Gene Therapy · doi:10.1038/gt.2011.200 · Cited by 48 · see citing works
- Authors
- Grimm, D., Pandey, K., Nakai, H., Storm, T. A., & Kay, M. A.
- Affiliations
- Stanford Medicine (US)
- Corresponding author
- Kay, Mark A.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2006-01
- Volume
- 80
- Issue
- 1
- Pages
- 426-439
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC1317553
- OpenAlex
- W2158777932
- Cited by
- 118
- References
- 12
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 6 · 2023: 5 · 2022: 8 · 2021: 5 · 2020: 4 · 2019: 8 · 2018: 4 · 2017: 9 · 2016: 3 · 2015: 7 · 2014: 8 · 2013: 4 · 2012: 4
- FWCI
- 3.78
- 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 (HL66948, U01HL066948)
- 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
- Health Sciences › Medicine › Hematology › Hemophilia Treatment and Research
- Keywords
- adeno-associated virus, inverted terminal repeats, liver gene transfer, hepatocyte transduction, factor IX, partial hepatectomy, portal vein infusion
- MeSH terms
- Dependovirus, Animals, Capsid, DNA, Viral, Genetic Vectors, HeLa Cells, Humans, Liver, Mice, Inbred C57BL, Recombination, Genetic, Transduction, Genetic, Mice
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