title: Long-term reversal of cholinergic neuronal decline in aged non-human primates by lentiviral NGF gene delivery.
abstract: Spontaneous atrophy of basal forebrain cholinergic neurons occurs with aging in the non-human primate brain. Short-term reversal of this atrophy has been reported following ex vivo nerve growth factor (NGF) gene delivery, but long-term effects of in vivo NGF gene delivery in the aged primate brain have not to date been examined. We tested the hypothesis that long-term lentiviral NGF intraparenchymal gene delivery would reverse age-related cholinergic decline, without induction of adverse effects previously observed following sustained intracerebroventricular growth factor protein exposure. Three aged rhesus monkeys underwent intraparenchymal lentiviral NGF gene delivery to the cholinergic basal forebrain. 1 year later, cholinergic neuronal numbers were quantified stereologically and compared to findings in four controls, non-treated aged monkeys and four young adult monkeys. Safety was assessed on several variables related to growth factor exposure. We now report that lentiviral gene delivery of NGF to the aged primate basal forebrain sustains gene expression for at least 1 year, and significantly restores cholinergic neuronal markers to levels of young monkeys. Aging resulted in a significant 17% reduction (p<0.05) in the number of neurons labeled for the cholinergic marker p75 among basal forebrain neurons. Lentiviral NGF gene delivery induced significant (p<0.05) and nearly complete recovery of p75-labeled neuronal numbers in aged subjects to levels observed in young monkeys. Similarly, the size of cholinergic neurons in aged monkeys was significantly reduced by 16% compared to young subjects (p<0.05), and lentiviral NGF delivery to aged subjects induced complete recovery of neuronal size. Intraparenchymal NGF gene delivery over a one-year period did not result in systemic leakage of NGF, activation of inflammatory markers in the brain, pain, weight loss, Schwann cell migration, or formation of anti-NGF antibodies. These findings indicate that extended trophic support to neurons in the non-human primate brain reverses age-related neuronal atrophy. These findings also support the safety and feasibility of lentiviral NGF gene transfer for potential testing in human clinical trials to protect degenerating cholinergic neurons in Alzheimer's disease.
authors.affiliation: Department of Neurosciences-0626, University of California-San Diego, La Jolla, California 92093, USA.
authors.forename: Alan H
authors.initials: AH
authors.lastname: Nagahara
authors.forename: Tim
authors.initials: T
authors.lastname: Bernot
authors.forename: Rod
authors.initials: R
authors.lastname: Moseanko
authors.forename: Laurie
authors.initials: L
authors.lastname: Brignolo
authors.forename: Armin
authors.initials: A
authors.lastname: Blesch
authors.forename: James M
authors.initials: JM
authors.lastname: Conner
authors.forename: Anthony
authors.initials: A
authors.lastname: Ramirez
authors.forename: Mehdi
authors.initials: M
authors.lastname: Gasmi
authors.forename: Mark H
authors.initials: MH
authors.lastname: Tuszynski
mesh_terms: D000109:Acetylcholine / Q000378:metabolism*; D000375:Aging / Q000473:pathology*; D000704:Analysis of Variance; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001284:Atrophy; D002452:Cell Count / Q000379:methods; D048429:Cell Size; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000378:metabolism; D000882:Haplorhini; D006801:Humans; D016086:Lentivirus / Q000502:physiology; D008279:Magnetic Resonance Imaging; D008297:Male; D020932:Nerve Growth Factor / Q000096:biosynthesis / Q000097:blood / Q000235:genetics / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects* / Q000502:physiology*; D016548:Prosencephalon / Q000473:pathology*; D020800:Receptor, Nerve Growth Factor / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D020800:Receptor, Nerve Growth Factor; D049452:Green Fluorescent Proteins; D020932:Nerve Growth Factor; D000109:Acetylcholine