Abstract
Stargardt disease (STGD1), due to mutations in the large ABCA4 gene, is the most common inherited macular degeneration in humans. Attempts at developing gene therapy approaches for treatment of STGD1 are currently ongoing. Among all the vectors available for gene therapy of inherited retinal diseases, those based on adeno-associated viruses (AAV) are the most promising given the efficacy shown in various animal models and their excellent safety profile in humans, as confirmed in many ongoing clinical trials. However, one of the main obstacles for the use of AAV is their limited effective packaging capacity of about 5 kb. Taking advantage of the AAV genome’s ability to concatemerize , others and we have recently developed dual AAV vectors to overcome this limit. We tested dual AAV vectors for ABCA4 delivery, and found that they transduce efficiently both mouse and pig photoreceptors , and rescue the Abca4−/− mouse retinal phenotype, indicating their potential for gene therapy of STGD1. This chapter details how we designed dual AAV vectors for the delivery of the ABCA4 gene and describes the techniques that can be explored to evaluate dual AAV transduction efficiency in vitro and in the retina, and their efficacy in the mouse model of STGD1.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Molday RS, Zhang K (2010) Defective lipid transport and biosynthesis in recessive and dominant Stargardt macular degeneration. Prog Lipid Res 49:476–492
Allikmets R, Singh N, Sun H et al (1997) A photoreceptor cell-specific ATP-binding transporter gene (ABCR) is mutated in recessive Stargardt macular dystrophy. Nat Genet 15:236–246
Trapani I, Puppo A, Auricchio A (2014) Vector platforms for gene therapy of inherited retinopathies. Prog Retin Eye Res 43:108–128
Dalkara D, Goureau O, Marazova K et al (2016) Let there be light: gene and cell therapy for blindness. Hum Gene Ther 27:134–147
Trapani I, Banfi S, Simonelli F et al (2015) Gene therapy of inherited retinal degenerations: prospects and challenges. Hum Gene Ther 26:193–200
Carvalho LS, Vandenberghe LH (2015) Promising and delivering gene therapies for vision loss. Vis Res 111:124–133
Colella P, Auricchio A (2012) Gene therapy of inherited retinopathies: a long and successful road from viral vectors to patients. Hum Gene Ther 23:796–807
Lipinski DM, Thake M, MacLaren RE (2013) Clinical applications of retinal gene therapy. Prog Retin Eye Res 32:22–47
Trapani I, Colella P, Sommella A et al (2014) Effective delivery of large genes to the retina by dual AAV vectors. EMBO Mol Med 6:194–211
Dyka FM, Boye SL, Chiodo VA et al (2014) Dual adeno-associated virus vectors result in efficient in vitro and in vivo expression of an oversized gene, MYO7A. Hum Gene Ther Methods 25:166–177
Lopes VS, Boye SE, Louie CM et al (2013) Retinal gene therapy with a large MYO7A cDNA using adeno-associated virus. Gene Ther 20:824–833
Colella P, Trapani I, Cesi G et al (2014) Efficient gene delivery to the cone-enriched pig retina by dual AAV vectors. Gene Ther 21:450–456
Yan Z, Zhang Y, Duan D et al (2000) Trans-splicing vectors expand the utility of adeno-associated virus for gene therapy. Proc Natl Acad Sci U S A 97:6716–6721
Duan D, Yue Y, Engelhardt JF (2001) Expanding AAV packaging capacity with trans-splicing or overlapping vectors: a quantitative comparison. Mol Ther 4:383–391
Ghosh A, Yue Y, Lai Y et al (2008) A hybrid vector system expands adeno-associated viral vector packaging capacity in a transgene-independent manner. Mol Ther 16:124–130
Trapani I, Colella P, Sommella A et al (2013) Effective delivery of large genes to the retina by dual AAV vectors. EMBO Mol Med 6:194–211
Ghosh A, Yue Y, Duan D (2011) Efficient transgene reconstitution with hybrid dual AAV vectors carrying the minimized bridging sequences. Hum Gene Ther 22:77–83
Auricchio A, Hildinger M, O'Connor E et al (2001) Isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column. Hum Gene Ther 12:71–76
Zhang Y, Chirmule N, Gao G et al (2000) CD40 ligand-dependent activation of cytotoxic T lymphocytes by adeno-associated virus vectors in vivo: role of immature dendritic cells. J Virol 74:8003–8010
Liang FQ, Anand V, Maguire AM et al (2001) Intraocular delivery of recombinant virus. Methods Mol Med 47:125–139
Mussolino C, della Corte M, Rossi S et al (2011) AAV-mediated photoreceptor transduction of the pig cone-enriched retina. Gene Ther 18:637–645
Weng J, Mata NL, Azarian SM et al (1999) Insights into the function of Rim protein in photoreceptors and etiology of Stargardt's disease from the phenotype in abcr knockout mice. Cell 98:13–23
Doria M, Ferrara A, Auricchio A (2013) AAV2/8 vectors purified from culture medium with a simple and rapid protocol transduce murine liver, muscle, and retina efficiently. Hum Gene Ther Methods. 24:392–398
Dong X, Tian W, Wang G et al (2010) Establishment of an AAV reverse infection-based array. PLoS One 5:e13479
Allocca M, Mussolino C, Garcia-Hoyos M et al (2007) Novel adeno-associated virus serotypes efficiently transduce murine photoreceptors. J Virol 81:11372–11380
Mattapallil MJ, Wawrousek EF, Chan CC et al (2012) The Rd8 mutation of the Crb1 gene is present in vendor lines of C57BL/6N mice and embryonic stem cells, and confounds ocular induced mutant phenotypes. Invest Ophthalmol Vis Sci 53:2921–2927
Gargiulo A, Bonetti C, Montefusco S et al (2009) AAV-mediated tyrosinase gene transfer restores melanogenesis and retinal function in a model of oculo-cutaneous albinism type I (OCA1). Mol Ther 17:1347–1354
Acknowledgments
Thanks to Raffaele Castello (Scientific Office, TIGEM, Pozzuoli, Italy) for the critical reading of this manuscript. Funding: The work was supported by the following: the European Research Council/ERC Grant agreement no282085 RetGeneTx; the European Community’s Seventh Framework Programme (FP7/2007–2013) under Grant agreement no 242013 Treatrush; the NIH (grant R24 EY019861-01A); the Italian Telethon Foundation (grant TGM11MT1).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Trapani, I. (2018). Dual AAV Vectors for Stargardt Disease. In: Boon, C., Wijnholds, J. (eds) Retinal Gene Therapy. Methods in Molecular Biology, vol 1715. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7522-8_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7522-8_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7521-1
Online ISBN: 978-1-4939-7522-8
eBook Packages: Springer Protocols