257. VEGF Gene Transfered Bone Marrow Stromal Cells by Replication-Deficiet Herpes Simplex Virus Type-1 (HSV-1) Vector Can Improve the Neurological Deficits and Reduce the Infaction in Rat Ischemic Brain

Recent animal studies demonstrated that some kind of growth factors can ameliorate neurological deficits and bone marrow stromal cells (MSCs) transplantation also can improve neurological function after ischemic stroke. MSCs, including the primitive pluriopotent mesenchmal stem cell, are attractive targets for cell and gene therapy. When long-term stable expression of a potentially therapeutic transgene is desired, as is the case for the treatment of cerebrovascular diseases, it may be appropriate to employ some viral vectors. The main advantage of the gene-engineered MSCs autotransplantation is that inadvertent gene transfer into other cell, including antigen- presenting cells, can be virtually excluded and the immunological interference can be get around, which is not the case in most in vivo gene therapy settings. The purpose of this study is to make sure if VEGF gene transfered MSC by using a replication-deficient HSV-1 vector can achieve further therapeutic effect against ischemic cerebral diseases than native MSC.Material and method:For ex vivo gene transduction to rat MSCs, we used a recombinant replication-deficient HSV-1 vector "1764/4-/pR19". This vector is disabled by the deletion of ICP4 and ICP34.5 and the inactivation of VP16 and contains the pR19 expression cassette driving a gene of interest under the control of fusion promoters,latency associated transcript promoter, cytomegalovirus enhancer, and woodchuck posttranscriptional regulatory elements. We had confirmed that this vector had a high ability of efficient gene transduction into MSCs with little cytotoxicity. We prepared the VEGF gene transfered MSCs ex vivo at MOI 5, and they were directly transplanted into the rat lesioned brain, 24 hours after transient middle cerebral artery occlusion (MCAO). As controls, we set the native MSCs group and PBS group. We used the modified neurological severity score (mNSS) to evaluate the neurological deficits and performed the 2,3,5-triphenyltetrazolium chloride (TTC) staining to check the infarct volume.Results:VEGF gene-transferred MSCs group showed the remarkable improvement of neurological symptoms and apparent reducing of infarct volume in comparison with native MSCs group and PBS group. We will present the detail data of this study.