title: High cerebrospinal fluid levels of interleukin-10 attained by AAV in dogs.
abstract: Intrathecal (IT) gene transfer using adeno-associated virus (AAV) may be clinically promising as a treatment for chronic pain if it can produce sufficiently high levels of a transgene product in the cerebrospinal fluid (CSF). Although this strategy was developed in rodents, no studies investigating CSF levels of an analgesic or antiallodynic protein delivered by IT AAV have been performed in large animals. Interleukin-10 (IL-10) is an antiallodynic cytokine for which target therapeutic levels have been established in rats. The present study tested IT AAV8 encoding either human IL-10 (hIL-10) or enhanced green fluorescent protein (EGFP) in a dog model of IT drug delivery. AAV8/hIL-10 at a dose of 3.5 × 10(12) genome copies induced high hIL-10 levels in the CSF, exceeding the target concentration previously found to be antiallodynic in rodents by >1000-fold. AAV8/EGFP targeted the primary sensory and motor neurons and the meninges. hIL-10, a xenogeneic protein in dogs, induced anti-hIL-10 antibodies detectable in the CSF and serum of dogs. The high hIL-10 levels demonstrate the efficacy of AAV for delivery of secreted transgenes into the IT space of large animals, suggesting a strong case for further development toward clinical testing.
authors.affiliation: Departments of Anesthesiology and Oncology, Mayo Clinic, Rochester, MN, USA.
authors.forename: J
authors.initials: J
authors.lastname: Pleticha
authors.affiliation: Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA.
authors.forename: S A
authors.initials: SA
authors.lastname: Malkmus
authors.affiliation: Departments of Anesthesiology and Oncology, Mayo Clinic, Rochester, MN, USA.
authors.forename: L F
authors.initials: LF
authors.lastname: Heilmann
authors.affiliation: Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA.
authors.forename: S L
authors.initials: SL
authors.lastname: Veesart
authors.affiliation: Departments of Anesthesiology and Oncology, Mayo Clinic, Rochester, MN, USA.
authors.forename: R
authors.initials: R
authors.lastname: Rezek
authors.affiliation: Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Q
authors.initials: Q
authors.lastname: Xu
authors.affiliation: Department of Anesthesiology, University of California, San Diego, La Jolla, CA, USA.
authors.forename: T L
authors.initials: TL
authors.lastname: Yaksh
authors.affiliation: Departments of Anesthesiology and Oncology, Mayo Clinic, Rochester, MN, USA.
authors.forename: A S
authors.initials: AS
authors.lastname: Beutler
mesh_terms: D000818:Animals; D059350:Chronic Pain / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics*; D004285:Dogs; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007278:Injections, Spinal; D016753:Interleukin-10 / Q000134:cerebrospinal fluid* / Q000235:genetics / Q000276:immunology; D008297:Male; D014161:Transduction, Genetic; D056189:Viral Tropism
chemical_list: C508609:IL10 protein, human; C485184:enhanced green fluorescent protein; D016753:Interleukin-10; D049452:Green Fluorescent Proteins