Regional Gene Therapy to Enhance Bone Repair

增强骨修复的区域基因治疗

• 批准号: 9899198
• 负责人: JAY R. LIEBERMAN
• 金额: $ 44.2万
• 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/9899198
• 关键词: Acute; 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; 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; side effect; soft tissue; stem cells; 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.

项目概要 该提案的目标是评估人类骨髓细胞的骨修复潜力 用含有 BMP-2 cDNA 的慢病毒载体转导的载体有多种。 没有一致令人满意的解决方案的困难骨修复场景包括： 骨折不愈合、急性骨折伴大量骨质流失、全关节置换术和 脊柱假关节传统上是自体骨移植的黄金标准，但 这种骨头的供应有限，人们担心相关的发病率 重组骨形态发生蛋白 (BMP) 已获得 FDA 批准使用。 然而，BMP 在脊柱融合和新鲜胫骨骨折治疗方面取得了不同程度的成功。 在人类中，与软组织水肿和异位等副作用有关 骨外科医生一直在寻找替代的组织工程。 我们的计划是开发区域基因疗法以增强骨修复。 转导骨髓细胞作为综合组织工程策略的一个方面 增强骨骼修复。 动物模型已证明，使用 BMP 进行基因治疗可产生骨髓 通过慢病毒基因转移产生的细胞有可能治疗中型或大型骨 缺陷或当宿主的生物环境受到严重损害时。 我们计划检验我们的假设，即用慢病毒载体转导的人骨髓细胞 过度表达 BMP-2 有可能增强人类骨骼的修复能力。 将从接受全髋关节置换术的患者的髓内管中采集骨髓细胞 关节成形术将用慢病毒载体（LV-TSTA-iC9/BMP-2）转导细胞。 含有 BMP-2 的 cDNA 和诱导性自杀基因 (iCaspase 9)，然后植入 在裸鼠的临界尺寸股骨缺损模型中，我们提出以下目标： 完成目标包括：1）比较骨后区域基因治疗的疗效 用新鲜分离或培养的扩增的人骨髓细胞进行修复；2) 建立 可以扩大用于人类的转导细胞的“细胞剂量”；以及 3) 确定 BMP 转导的人体细胞的局部和全身生物分布并评估器官 毒性、载体遗传毒性和异位骨化从该提案中获得的数据。 将提供有关调整该区域基因治疗策略作为 人类的治疗方案。