Zinc finger protein transcription factors (ZFP TFs) can be engineered to regulate expression of any endogenous target gene, and thus represent a novel ...
CENTRAL NERVOUS SYSTEM II tissue(s). These results suggest that NVV-based IL-10 gene therapy may be a potent addition to the treatments available to neuropathic pain patients.
531. Activation of VEGF by Engineered Zinc Finger Transcriptional Activators: A Novel Therapeutic Approach to Diabetic Neuropathy Carolyn L. Dent,1 Gabriel Lau,1 Edward J. Rebar,1 Lei Zhang,1 YaLi Lee,1 Andrew C. Jamieson,1 Casey C. Case,1 Tyler Martin,1 Philip D. Gregory.1 1 Sangamo Biosciences Inc., Richmond, CA. Zinc finger protein transcription factors (ZFP TFs) can be engineered to regulate expression of any endogenous target gene, and thus represent a novel approach to the treatment of disease. In the area of diabetic neuropathy, for example, where no current treatment options are available, ZFP TF technology offers a unique solution to an unmet medical need. Diabetic neuropathy is a progressive nerve degeneration that occurs in over half of patients with diabetes (1). In a phase I/II clinical trial of VEGF165 gene therapy for critical limb ischemia, in addition to improvements in angiogenesis and blood flow, anecdotal neurological improvements were also observed in patients with diabetes(2). Moreover, VEGF165 has demonstrated clear direct neurotrophic and neuroprotective effects on neurons and Schwann cells in culture. In a relevant animal model extensive nerve damage such as loss of axons and demyelination was reversed when the animals were treated with VEGF165 delivered as a plasmid gene therapy either at the time of injury or 10 days post-injury (3). A direct stimulatory effect on the migration and survival of isolated Schwann cells in culture was also demonstrated in this work. Others have shown that VEGF directly stimulates the outgrowth of neurites from DRG and sciatic nerve explants, and has a protective effect against death as a result of serum starvation in neuronal cell lines (4,5). Up-regulation of VEGF by a ZFP TF thus appears to be a potential strategy for the treatment of diabetic neuropathy. We have designed a ZFP TF that induces expression of the endogenous VEGF gene, and thus of all the splice variants thought to be necessary for normal cellular function. We have shown that we can up-regulate the endogenous VEGF gene in multiple cell lines, including neurons, using a ZFP TF. VEGF produced this way can then protect neuronal cells from damage occurring as a result of diabetic neuropathy. (1) Feldman E (2003) J. Clin. Invest. 111:431-433 (2) Simovic MD et al (2001) Arch. Neurol. 58:761-768 (3) Schratzberger P et al (2000) Nature Med. 6:405-413 (4) Sondell M et al (1999) J. Neurosci. 19:5731-5740 (5) Jin KL et al (2000) J. Mol. Neurosci. 14:197-203
532. Cervical Spinal Cord Delivery of a Lentiviral Vector in SOD-1 Transgenic Mice Diana Tanase,1 Qingshan Teng,1 Ajit A. Krishnaney,1 James K. Liu,1 Mary E. Garrity-Moses,1 Nicholas Mazarakis,2 Lucy E. Walmsley,2 Mimoun Azzouz,2 Nicholas M. Boulis.1 1 Neurological Surgery, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH; 2Oxford BioMedica, Medawar Centre, Oxford, United Kingdom. Objective: Recent work has suggested that injection of lentiviral vectors might constitute means of therapeutic gene delivery to diseased motor neurons in ALS. However, as implied by the name, Amyotrophic Lateral Sclerosis, this disorder results in sclerosis of the spinal cord. In addition, axonal transport is altered in these animals. The present study attempts to characterize the behavioral impact as well as the distribution of gene expression in the CNS Molecular Therapy Volume 9, Supplement 1, May 2004 Copyright The American Society of Gene Therapy
following direct spinal cord injection of the rabies G protein pseudotyped lentiviral vector, EIAV.LacZ, in control and SOD-1 transgenic mice. Methods: The BBB locomotor score, grip strength meter, and rotarod assays were used to assess disease progression and the behavioral effects of cervical spinal cord microinjection of the viral vector RabG.EIAV.LacZ in SOD-1 transgenic and wild-type control mice. Peripheral and central nervous systems staining for the reporter gene β-galactosidase were histologically quantified after microinjection of RabG.EIAV.LacZ into cervical spinal cord, brachial plexus or deltoid muscle of mice. Results: Direct cervical spinal cord microinjection of RabG.EIAV.lacZ does not affect motor performance as compared to vehicle injection as determined by three different methods. Vector injection and transgene expression does not accelerate disease progression. In addition, direct cervical spinal cord RabG.EIAV.lacZ injection results in extensive CNS uptake in SOD-1 transgenic mice, statistically similar to wild type mice (p>0.05). Direct brachial plexus injection results in significantly higher CNS uptake than intramuscular injections (p
