title: Widespread and efficient transduction of spinal cord and brain following neonatal AAV injection and potential disease modifying effect in ALS mice.
abstract: The architecture of the spinal cord makes efficient delivery of recombinant adeno-associated virus (rAAV) vectors throughout the neuraxis challenging. We describe a paradigm in which small amounts of virus delivered intraspinally to newborn mice result in robust rAAV-mediated transgene expression in the spinal cord. We compared the efficacy of rAAV2/1, 2/5, 2/8, and 2/9 encoding EGFP delivered to the hindlimb muscle (IM), cisterna magna (ICM), or lumbar spinal cord (IS) of neonatal pups. IS injection of all four capsids resulted in robust transduction of the spinal cord with rAAV2/5, 2/8, and 2/9 vectors appearing to be transported to brain. ICM injection resulted in widespread expression of EGFP in the brain, and upper spinal cord. IM injection resulted in robust muscle expression, with only rAAV2/8 and 2/9 transducing spinal motor and sensory neurons. As proof of concept, we use the IS paradigm to express murine Interleukin (IL)-10 in the spinal cord of the SOD1-G93A transgenic mouse model of amyotrophic lateral sclerosis. We show that expression of IL-10 in the spinal axis of SOD1-G93A mice altered the immune milieu and significantly prolonged survival. These data establish an efficient paradigm for somatic transgene delivery of therapeutic biologics to the spinal cord of mice.
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Jacob I
authors.initials: JI
authors.lastname: Ayers
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Fromholt
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Olga
authors.initials: O
authors.lastname: Sinyavskaya
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Zoe
authors.initials: Z
authors.lastname: Siemienski
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Awilda M
authors.initials: AM
authors.lastname: Rosario
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Li
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Keith W
authors.initials: KW
authors.lastname: Crosby
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Pedro E
authors.initials: PE
authors.lastname: Cruz
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Nadia M
authors.initials: NM
authors.lastname: DiNunno
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Janus
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Carolina
authors.initials: C
authors.lastname: Ceballos-Diaz
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Borchelt
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Todd E
authors.initials: TE
authors.lastname: Golde
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Paramita
authors.initials: P
authors.lastname: Chakrabarty
authors.affiliation: Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease and McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, Florida, USA.
authors.forename: Yona
authors.initials: Y
authors.lastname: Levites
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D002213:Capsid / Q000378:metabolism; D002946:Cisterna Magna / Q000378:metabolism / Q000473:pathology; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007278:Injections, Spinal; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology; D013116:Spinal Cord / Q000378:metabolism* / Q000473:pathology; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: C556062:IL10 protein, mouse; C485184:enhanced green fluorescent protein; D016753:Interleukin-10; D049452:Green Fluorescent Proteins; C109415:SOD1 G93A protein; D013482:Superoxide Dismutase