title: Transcriptional Response of Human Neurospheres to Helper-Dependent CAV-2 Vectors Involves the Modulation of DNA Damage Response, Microtubule and Centromere Gene Groups.
abstract: Brain gene transfer using viral vectors will likely become a therapeutic option for several disorders. Helper-dependent (HD) canine adenovirus type 2 vectors (CAV-2) are well suited for this goal. These vectors are poorly immunogenic, efficiently transduce neurons, are retrogradely transported to afferent structures in the brain and lead to long-term transgene expression. CAV-2 vectors are being exploited to unravel behavior, cognition, neural networks, axonal transport and therapy for orphan diseases. With the goal of better understanding and characterizing HD-CAV-2 for brain therapy, we analyzed the transcriptomic modulation induced by HD-CAV-2 in human differentiated neurospheres derived from midbrain progenitors. This 3D model system mimics several aspects of the dynamic nature of human brain. We found that differentiated neurospheres are readily transduced by HD-CAV-2 and that transduction generates two main transcriptional responses: a DNA damage response and alteration of centromeric and microtubule probes. Future investigations on the biochemistry of processes highlighted by probe modulations will help defining the implication of HD-CAV-2 and CAR receptor binding in enchaining these functional pathways. We suggest here that the modulation of DNA damage genes is related to viral DNA, while the alteration of centromeric and microtubule probes is possibly enchained by the interaction of the HD-CAV-2 fibre with CAR.
authors.affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Piersanti
authors.affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy.
authors.forename: Romina
authors.initials: R
authors.lastname: Burla
authors.affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy; Pasteur Institute, Cenci Bolognetti Foundation, Rome, Italy.
authors.forename: Valerio
authors.initials: V
authors.lastname: Licursi
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157, Oeiras, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Brito
authors.affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy.
authors.forename: Mattia
authors.initials: M
authors.lastname: La Torre
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157, Oeiras, Portugal.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157, Oeiras, Portugal.
authors.forename: Daniel
authors.initials: D
authors.lastname: Simao
authors.affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Mottini
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS UMR 5535, Montpellier, France; Université de Montpellier, Montpellier, France.
authors.forename: Sara
authors.initials: S
authors.lastname: Salinas
authors.affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy; Pasteur Institute, Cenci Bolognetti Foundation, Rome, Italy.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Negri
authors.affiliation: Department of Biomedical Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Tagliafico
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS UMR 5535, Montpellier, France; Université de Montpellier, Montpellier, France.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Kremer
authors.affiliation: Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, Rome, Italy; Pasteur Institute, Cenci Bolognetti Foundation, Rome, Italy; Institute of Molecular Biology and Pathology, CNR, Rome, Italy.
authors.forename: Isabella
authors.initials: I
authors.lastname: Saggio
mesh_terms: D006516:Adenoviruses, Canine / Q000502:physiology*; D000818:Animals; D001921:Brain / Q000378:metabolism; D002478:Cells, Cultured; D002503:Centromere / Q000235:genetics* / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics*; D004285:Dogs; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D008870:Microtubules / Q000235:genetics* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism*; D019076:Transgenes / Q000502:physiology*