Regional Gene Therapy to Enhance Bone Repair

增强骨修复的区域基因治疗

• 批准号: 9238335
• 负责人: JAY R. LIEBERMAN
• 金额: $ 60.48万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9238335
• 关键词: Acute; Adverse effects; Animal Model; Apoptosis; Autologous; Biodistribution; Biological; Bone Marrow; Bone Marrow Cells; Bone Morphogenetic Proteins; Bone Regeneration; Bone Transplantation; Cell Count; Cells; Cessation of life; Clinical; Clinical Data; Complementary DNA; Data; Defect; Dose; Edema; Environment; FDA approved; Fracture; Funding; Gene Transfer; Genes; Goals; Gold; Harvest; Heterotopic Ossification; Histologic; Human; Immune response; Implant; Knowledge; Laboratories; Lentivirus Vector; Longitudinal Studies; Methods; Modeling; Morbidity - disease rate; Mus; Nude Rats; Organ; Orthopedics; Patients; Procedures; Proteins; Pseudarthrosis; Recombinants; Regimen; Replacement Arthroplasty; Research; Risk; Safety; Site; Spinal Fusion; Stem cells; Surgeon; TSTA; Testing; Tibial Fractures; Tissue Engineering; Toxic effect; Treatment Protocols; Vertebral column; Viral Genome; Work; base; bone; bone cell; bone loss; bone morphogenetic protein 2; bone quality; cellular transduction; comparative efficacy; experimental study; gene therapy; genetic manipulation; genotoxicity; healing; hip replacement arthroplasty; implantation; overexpression; pre-clinical; programs; scale up; soft tissue; success; suicide gene; tissue culture; vector

Project Summary The goal of this proposal is to assess the bone repair potential of human bone marrow cells transduced with a lentiviral vector containing the cDNA for BMP-2. There are a number of difficult bone repair scenarios for which there is no consistently satisfactory solution including: fracture nonunion, acute fractures with extensive bone loss, revision total joint arthroplasty and pseudarthrosis of the spine. Traditionally, autologous bone graft has been the gold standard but there is a limited supply of this bone and there are concerns regarding the morbidity associated with graft harvest. Recombinant bone morphogenetic proteins (BMP) are FDA approved for use in spinal fusion and treatment of fresh tibial fractures. However, BMPs have had mixed success in humans and are associated with side effects including soft tissue edema and heterotopic ossification. Orthopaedic surgeons have been searching for alternative tissue engineering strategies to enhance bone repair. Our plan is to develop regional gene therapy using transduced bone marrow cells as one aspect of a comprehensive tissue engineering strategy to enhance bone repair. It has been demonstrated in animal models that gene therapy with BMP producing bone marrow cells created via lentiviral gene transfer has the potential to treat either medium or large bone defects or when the biological environment of the host is severely compromised. In this proposal we plan to test our hypothesis that human bone marrow cells transduced with a lentiviral vector to overexpress BMP-2 have the potential to enhance bone repair in humans. Human bone marrow cells will be harvested from the intramedullary canals of patients undergoing total hip arthroplasty. The cells will be transduced with a lentiviral vector (LV-TSTA-iC9/BMP-2) that contains the cDNA for BMP-2 and an inducible suicide gene (iCaspase 9) and then implanted in a critical sized femoral defect model in nude rats. We propose the following aims to accomplish the goal including: 1) compare the efficacy of regional gene therapy after bone repair with freshly isolated or cultured expanded human bone marrow cells; 2) establish a “cellular dose” of transduced cells that can be scaled up for use in humans; and 3) determine the local and systemic biodistribution of BMP transduced human cells and assess for organ toxicity, vector genotoxicity and heterotopic ossification. The data obtained from this proposal will provide critical information with respect to adapting this regional gene therapy strategy as a treatment regimen in humans.

项目摘要 这项提案的目的是评估人类骨髓细胞的骨修复潜力 用含有BMP-2的cDNA的慢病毒载体转导。有许多 没有一致满意的解决方案的困难骨修复情况包括： 骨折不愈合，急性骨折伴广泛骨丢失，翻修型全关节置换术， 脊柱假关节。传统上，自体骨移植一直是金标准， 这种骨的供应有限， 移植物收获。重组骨形态发生蛋白（BMP）是FDA批准使用 脊柱融合术和新鲜胫骨骨折的治疗。然而，BMP的成功与否好坏参半 并且与包括软组织水肿和异位的副作用有关 骨化整形外科医生一直在寻找替代组织工程 增强骨修复的策略。我们的计划是开发区域性基因疗法， 转导的骨髓细胞作为综合组织工程策略的一个方面， 增强骨修复。 在动物模型中已经证明，用产生骨髓的BMP进行基因治疗， 通过慢病毒基因转移产生的细胞具有治疗中等或大骨的潜力 缺陷或当宿主的生物环境严重受损时。本提案中 我们计划验证我们的假设，即用慢病毒载体转导的人骨髓细胞 过表达BMP-2具有增强人类骨修复的潜力。人骨 将从接受全髋关节置换术的患者的髓内管中收获骨髓细胞 关节成形术将用慢病毒载体（LV-TSTA-iC 9/BMP-2）转导细胞， 含有BMP-2的cDNA和诱导性自杀基因（iCaspase 9）， 在裸大鼠的临界尺寸股骨缺损模型中。我们提出以下目标， 完成的目标包括：1）比较骨转移后区域基因治疗的疗效 用新鲜分离的或培养的扩增的人骨髓细胞修复; 2）建立 可以按比例放大用于人类的转导细胞的“细胞剂量”;以及 BMP转导人细胞的局部和全身生物分布以及对器官的评估 毒性、载体遗传毒性和异位骨化。从该提案中获得的数据 将提供关于适应这种区域基因治疗策略的关键信息， 治疗方案。