title: Evaluation of helper-dependent canine adenovirus vectors in a 3D human CNS model.
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.
authors.affiliation: iBET-Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: D
authors.initials: D
authors.lastname: Simão
authors.affiliation: iBET-Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: C
authors.initials: C
authors.lastname: Pinto
authors.affiliation: iBET-Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: P
authors.initials: P
authors.lastname: Fernandes
authors.affiliation: The Francis Crick Institute, Lincoln's Inn Fields Laboratory, London, UK.
authors.forename: C J
authors.initials: CJ
authors.lastname: Peddie
authors.affiliation: Dipartimento di Biologia e Biotecnologie 'Charles Darwin', Università di Roma La Sapienza, Rome, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Piersanti
authors.affiliation: The Francis Crick Institute, Lincoln's Inn Fields Laboratory, London, UK.
authors.forename: L M
authors.initials: LM
authors.lastname: Collinson
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, Montpellier, France.|Université Montpellier, Montpellier, France.
authors.forename: S
authors.initials: S
authors.lastname: Salinas
authors.affiliation: Dipartimento di Biologia e Biotecnologie 'Charles Darwin', Università di Roma La Sapienza, Rome, Italy.|Istituto Pasteur Fondazione Cenci Bolognetti, Università di Roma La Sapienza, Rome, Italy.|Istituto di Biologia e Patologia Molecolari del CNR, Università di Roma La Sapienza, Rome, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Saggio
authors.affiliation: Sobell Department of Motor Neuroscience and Movement Disorders, Institute of Neurology, University College London, London, UK.
authors.forename: G
authors.initials: G
authors.lastname: Schiavo
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, Montpellier, France.|Université Montpellier, Montpellier, France.
authors.forename: E J
authors.initials: EJ
authors.lastname: Kremer
authors.affiliation: iBET-Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: C
authors.initials: C
authors.lastname: Brito
authors.affiliation: iBET-Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: P M
authors.initials: PM
authors.lastname: Alves
mesh_terms: D006516:Adenoviruses, Canine / Q000235:genetics*; D000260:Adenoviruses, Human / Q000235:genetics; D000818:Animals; D001370:Axonal Transport; D002454:Cell Differentiation; D002470:Cell Survival; D002490:Central Nervous System / Q000378:metabolism*; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes; D056189:Viral Tropism