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Gene Ther. 2016 Jan;23(1):86-94. doi: 10.1038/gt.2015.75. Epub 2015 Aug 20.

Evaluation of helper-dependent canine adenovirus vectors in a 3D human CNS model.

Author information

1
iBET-Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
2
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
3
The Francis Crick Institute, Lincoln's Inn Fields Laboratory, London, UK.
4
Dipartimento di Biologia e Biotecnologie 'Charles Darwin', Università di Roma La Sapienza, Rome, Italy.
5
Institut de Génétique Moléculaire de Montpellier, Montpellier, France.
6
Université Montpellier, Montpellier, France.
7
Istituto Pasteur Fondazione Cenci Bolognetti, Università di Roma La Sapienza, Rome, Italy.
8
Istituto di Biologia e Patologia Molecolari del CNR, Università di Roma La Sapienza, Rome, Italy.
9
Sobell Department of Motor Neuroscience and Movement Disorders, Institute of Neurology, University College London, London, UK.

Abstract

Gene therapy is a promising approach with enormous potential for treatment of neurodegenerative disorders. Viral vectors derived from canine adenovirus type 2 (CAV-2) present attractive features for gene delivery strategies in the human brain, by preferentially transducing neurons, are capable of efficient axonal transport to afferent brain structures, have a 30-kb cloning capacity and have low innate and induced immunogenicity in preclinical tests. For clinical translation, in-depth preclinical evaluation of efficacy and safety in a human setting is primordial. Stem cell-derived human neural cells have a great potential as complementary tools by bridging the gap between animal models, which often diverge considerably from human phenotype, and clinical trials. Herein, we explore helper-dependent CAV-2 (hd-CAV-2) efficacy and safety for gene delivery in a human stem cell-derived 3D neural in vitro model. Assessment of hd-CAV-2 vector efficacy was performed at different multiplicities of infection, by evaluating transgene expression and impact on cell viability, ultrastructural cellular organization and neuronal gene expression. Under optimized conditions, hd-CAV-2 transduction led to stable long-term transgene expression with minimal toxicity. hd-CAV-2 preferentially transduced neurons, whereas human adenovirus type 5 (HAdV5) showed increased tropism toward glial cells. This work demonstrates, in a physiologically relevant 3D model, that hd-CAV-2 vectors are efficient tools for gene delivery to human neurons, with stable long-term transgene expression and minimal cytotoxicity.

PMID:
26181626
PMCID:
PMC5367519
DOI:
10.1038/gt.2015.75
[Indexed for MEDLINE]
Free PMC Article

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