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Mol Ther. 2018 Feb 7;26(2):524-541. doi: 10.1016/j.ymthe.2017.11.019. Epub 2017 Dec 5.

Triple Vectors Expand AAV Transfer Capacity in the Retina.

Author information

1
Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli 80078, Italy.
2
Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli 80078, Italy; Armenise/Harvard Laboratory of Integrative Genomics, TIGEM, Pozzuoli 80078, Italy.
3
Eye Clinic, Multidisciplinary Department of Medical, Surgical and Dental Sciences, Second University of Naples, Naples 80121, Italy.
4
The Jackson Laboratory, Bar Harbor, ME 04609, USA.
5
Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli 80078, Italy; Medical Genetics, Department of Advanced Biomedicine, Federico II University, Naples 80131, Italy. Electronic address: auricchio@tigem.it.

Abstract

Retinal gene transfer with adeno-associated viral (AAV) vectors holds great promise for the treatment of inherited retinal degenerations (IRDs). One limit of AAV is its transfer capacity of about 5 kb, which can be expanded to about 9 kb, using dual AAV vectors. This strategy would still not suffice for treatment of IRDs such as Usher syndrome type 1D or Alström syndrome type I (ALMS) due to mutations in CDH23 or ALMS1, respectively. To overcome this limitation, we generated triple AAV vectors, with a maximal transfer capacity of about 14 kb. Transcriptomic analysis following triple AAV transduction showed the expected full-length products along a number of aberrant transcripts. However, only the full-length transcripts are efficiently translated in vivo. We additionally showed that approximately 4% of mouse photoreceptors are transduced by triple AAV vectors and showed correct localization of recombinant ALMS1. The low-photoreceptor transduction levels might justify the modest and transient improvement we observe in the retina of a mouse model of ALMS. However, the levels of transduction mediated by triple AAV vectors in pig retina reached 40% of those observed with single vectors, and this bodes well for further improving the efficiency of triple AAV vectors in the retina.

KEYWORDS:

AAV; gene therapy; large gene; retina; triple AAV

PMID:
29292161
PMCID:
PMC5835116
DOI:
10.1016/j.ymthe.2017.11.019
[Indexed for MEDLINE]
Free PMC Article

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