Marco A. Passini,1 Jie Bu,1 Jonathan A. Fidler,1 Joseph W. Foley,1 James C. Dodge,1 Robin J. Ziegler,1 Wendy W. Yang,1. Jennifer Clarke,1 Tatyana V. Taksir ...
NEUROLOGIC sequence-specific binding platform for cellular splicing factors, such as SR proteins. Plasmid-based and recombinant adeno-associated virus vectors were developed that expressed bi-functional RNAs that stimulated SMN2 exon 7 inclusion and full-length SMN protein in patient fibroblasts. These experiments provide a mechanism to modulate splicing from a variety of genetic contexts and directly demonstrate a novel therapeutic approach for SMA.
416. Gene Therapy for Parkinsons Disease by In Vivo Plasmid Transfer of Human Hepatocyte Growth Factor in Primate Model Akihiko Ishida, Hiromi Koike, Ryuichi Morishita. 1 Division of Clinical Gene Therapy, School of Medicine, Osaka University, Suita, Osaka, Japan. Parkinson’s disease (PD) is a neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra. The ultimate treatment of PD is to protect and repair dopaminergic neurons from degenerative process. As neurotrophic factors have been shown to support the survival and enhance the function of dopaminergic neurons, gene therapy using neurotrophic factors such as glial cell line-derived neurotrophic factor becames center of interests. In the present study, we focused on hepatocyte growth factor (HGF) as a novel neurotrophic and angiogenic growth factor that is a well-known potent pleiotrophic cytokine exhibiting mitogenic, motogenic, and morphogenic activities in a variety of cells. Prevention study: we have extended the preclinical exploration to primate model of PD. Seven days after stereotaxic transfection of human HGF plasmid or lacZ plasmid into the unilateral striatum, infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) into the right internal carotid artery of the monkey produces toxininduced injury to the right nigro-striatal pathway. After MPTP infusion for 4 weeks, control animals exhibited stable moderate parkinsonian features. However, primates transfecterd with HGF plasmid showed normal states. PD model primates transfected with lacZ plasmid demonstrated the apomorphine and amphetamineinduced rotational asymmetry. On the other hand, transfection of human HGF plasmid resulted in a significant inhibition of abnormal rotation for 6 months. Microdialysis demonstrated that concentrations of dopamine in striatum and substantia nigra were higher level compared with PD model and lacZ plasmid transfected animals. PET imaging demonstrated remarkable and persistent increase of [11C]-L-dopa uptake and CFT binding in HGF plasmid injected striatum, while [11C]-raclopride did not change at the HGF plasmid terated site. Overall, the present study demonstrated that over-expression of human HGF prevented neuronal death in PD model. Repair study: Primate models which infused MPTP into the right internal carotid artery ,exhibited stable moderate parkinsonian features. PD primates were stereotaxic transfection of human HGF plasmid or lacZ plasmid was performed into the unilateral striatum. Primates transfecterd with HGF plasmid into striatum showed to improved normal states gradually. PD model primates transfected with lacZ plasmid demonstrated the apomorphine and amphetamineinduced rotational asymmetry. On the other hand, transfection of human HGF plasmid resulted in a almost inhibition of abnormal rotation for over 6 months. Doparminergic neurons of PD model primates transfected with human HGF plasmid into striatum were repaired significantly as assessed by immunohistchemistry. The present study demonstrated that over-expression of human HGF repaired neuronal degeneration and improved of symptoms in PD model. These two studies results indicate that HGF gene therapy may represent a promising candidate treatment for PD. S160
417. Combination Brain and Systemic Injections of AAV Results in Whole Body Therapy and Extension of Lifespan in the Niemann-Pick Mouse Marco A. Passini,1 Jie Bu,1 Jonathan A. Fidler,1 Joseph W. Foley,1 James C. Dodge,1 Robin J. Ziegler,1 Wendy W. Yang,1 Jennifer Clarke,1 Tatyana V. Taksir,1 Denise A. Griffiths,1 Michael A. Zhao,1 Catherine R. O’Riordan,1 Lamya S. Shihabuddin,1 Edward H. Schuchman,2 Seng H. Cheng.1 1 Neuroscience, Genzyme Corporation, Framingham, MA; 2 Human Genetics, Mount Sinai School of Medicine, New York, NY. The majority of lysosomal storage diseases contain both CNS and visceral pathology. Many experimental designs use either intracranial injection of viral vectors to treat the neurodegenerative phenotype of the disease, or systemic delivery to treat the viscera. In this study, we tested whether a combination of brain and systemic injections of adeno-associated virus (AAV) vectors could increase lifespan, provide whole body reversal of pathology, and improve motor and cognitive function in the acid sphingomyelinase knock out (ASMKO) mouse model of Niemann-Pick disease. ASMKO mice at 4 weeks of age were injected systemically with AAV8hASM, and subsequently, the same animals were injected into the brain with AAV2-hASM. All the mice treated by combination injections survived to 54 weeks of age. This was a significant improvement over untreated ASMKO mice, animals that had been treated by systemic injection only or by brain injection only, which had median life spans of 34 weeks, 45 weeks, and 43 weeks, respectively. Thus the brain alone injections did not protect animals from dying. Animals treated by the combination therapy also displayed normal weight gain, and significant functional recovery on the rotarod (motor task) and the Barnes maze (cognitive task) throughout the time course of the study. These data support the contention that combination therapy can improve the quality of life before and within the time frame of extended survival.
418. AAV-Mediated sFLT-1 Gene Therapy for Ocular Neovascularization: Longevity, Toxicity and Efficacy Results from Mice and Monkeys E. P. Rakoczy,1,2 C. M. Lai,1,2 W. Y. Shen,1,2 M. Brankov,1,2 N. Barnett,2,3 S. Y. Lee,4 S. G. Leaver,2 I. Yeo,4 C. L. Ang,4 I. J. Constable.1,2 1 Centre for Ophthalmology and Visual Science, The University of Westerna Australia, Perth, Australia; 2Molecular Ophthalmology, The Lions Eye Institute, Perth, Australia; 3Vision, Touch and Hearing Research Centre, The University of Queensland, Brisbane, QLD, Australia; 4Singapore National Eye Centre, Singapore. Purpose: To investigate the efficacy, longevity and toxicity of adeno-associated virus (AAV)-mediated expression of sFlt-1 in the mouse model of retinal neovascularization (trVEGF029) and in the laser induced monkey model of choroidal neovascularization (CNV). Methods: rAAV.sFLT was injected subretinally into trVEGF029 mouse (n=26) and monkey (n=4) eyes. 16 months post-injection of rAAV CNV was induced in monkey eyes by laser photocoagulation. Fluorescence angiography, histology and immunological techniques were used to measure the efficacy, longevity and toxicity of the viral construct. Results: Transgene expression was maintained for at least 8 and 17 months post-injection in mice and monkeys, respectively. The expression of sFlt-1 was associated with the longterm (8 months) regression of neovascular vessels in 85% of trVEGF029 eyes. The majority of the treated trVEGF029 eyes (75%) retained high numbers of photoreceptors. AAV-mediated expression of sFlt-1 prevented the development of laser photocoagulation-induced CNV in all treated monkey eyes. There Molecular Therapy Volume 13, Supplement 1, May 2006 Copyright The American Society of Gene Therapy
