MARROW STROMAL CELLS IN OSTEOGENESIS IMPERFECTA MODEL

描述(申请人提供)：成骨不全(OI) 1/10,000-1/20,000活产儿的发病率，经常被作为结构蛋白显性阴性条件的典范疾病。有效 对这些疾病的治疗将需要直接或有针对性的替换 野生型等位基因突变。这项研究计划将探索 OI患者有可能通过自己的方式治疗 骨祖细胞通过基因工程从他们的骨髓中提取的细胞 称为间充质干细胞或骨髓基质细胞(MSCs)。这个 骨髓基质细胞将被用来检验骨髓基质的假设 来自OI患者的细胞可以通过基因工程来纠正 导致这种疾病的I型胶原突变的有害影响。 这一提议的具体目的是：(1)获得同源重组 COL1A1基因在人骨髓间充质干细胞中的表达，以确定缓解 用COL1A1野生型等位基因替换突变等位基因引起的OI症状 (2)获得高表达COL1A1基因的hMSCs克隆。 测定野生型与突变型COL1A1蛋白的比例对 基因工程人骨髓间充质干细胞的修复潜能及归巢研究 基因工程MSCs的潜力将通过注射细胞来确定 成骨折模型和对照组小鼠。

DESCRIPTION (provided by applicant): Osteogenesis Imperfecta (OI) that has an incidence of 1/10,000 - 1/20,000 live births, has frequently served as a model disorder for dominant negative conditions of structural proteins. Effective therapy for these disorders will require direct or targeted replacement of a mutated gene with wild-type allele. This research proposal will explore the possibility that patients with OI can potentially be treated with their own osteoprogenitors by gene-engineering the cells from their bone marrow that are referred to as mesenchymal stem cells or marrow stromal cells (MSCs). The marrow stromal cells will be used to test the hypothesis that marrow stromal cells from the patients with OI can be gene-engineered to correct the deleterious effects of mutations in type I collagen that produce the disease. The Specific Aims for this proposal are: (1) To obtain homologous recombination of the COL1A1 gene in human MSCs to determine the possibility of alleviating the OI symptoms by replacing the mutant allele with wild type allele of COL1A1 (2) To obtain over-expression clones of hMSCs that express COL1A1 cDNA to determine the effect of ratio of wild-type to mutant COL1A1 protein (3) To study the repair potential and homing of gene engineered hMSCs; the repair potential of gene engineered MSCs will be determined by injecting the cells into the fracture model and control mice.