AAV-Mediated Gene Therapy for Metabolic Bone Disease

AAV 介导的代谢性骨病基因治疗

基本信息

批准号：
7676053
负责人：
Selvarangan Ponnazhagan
金额：
$ 26.77万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-15 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7676053
关键词：
Adverse effects Affect Anabolic Agents Area Autologous Transplantation BMP2 gene Biodistribution Bone Density Bone Development Bone Marrow Bone Morphogenetic Proteins Bone Resorption Bone remodeling Cell Transplants Cells Collagen Type I Defect Dependovirus Development Disease Ectopic Expression Engraftment Etiology Event Fracture Functional disorder Future Gene Transfer Genetic Half-Life Hematologic Neoplasms Histology Home environment Homing Hormonal Human Image Immunocompetent Immunohistochemistry In Vitro Integrins Lead Luc Gene Mediating Mesenchymal Stem Cells Metabolic Metabolic Bone Diseases Metabolic Diseases Methods Mus Mutation Osteoblasts Osteoclasts Osteogenesis Osteogenesis Imperfecta Osteoporosis Outcome Study Pathogenicity Pharmaceutical Preparations Pharmacotherapy Production Proliferating Property Proteins Recombinant adeno-associated virus (rAAV)Recombinants Reporter Signal Transduction Source Spinal Fusion Stromal Cells Testing Therapeutic Effect Tissues Transgenes Viral Genes adeno-associated viral vector age related base bone bone mass bone morphogenetic protein 2 cancer cell gene therapy human disease immunogenic immunogenicity improved in vivo mouse model novel novel therapeutics older patient osteoblast differentiation osteogenic osteopontin osteoprogenitor cell pre-clinical prevent promoter research clinical testing self-renewal senescence spine bone structure therapeutic gene transgene expression vector

项目摘要

DESCRIPTION (provided by applicant): Adeno-associated virus (AAV) vectors are ideal for the long-term treatment of metabolic defects. Unique features including non-pathogenicity, low-immunogenicity and stable expression have positively resulted in successful preclinical and clinical evaluation of this vector. One of the potential, yet, unexplored areas of gene therapy using rAAV is metabolic bone defects characterized by a reduction in the mass of bone per unit volume as in osteoporosis or insufficient mechanisms for anabolic bone remodeling as in spinal fusion and fracture. Although, currently available hormonal and drug therapies for osteopenia aim to prevent further bone destruction by osteoclasts, therapies directed towards increasing bone mass by increasing the event of osteogenesis will be greatly beneficial. Treatments to increase bone density by anabolic agents are limited due to ineffective delivery methods and a short half-life of the drugs and purified proteins. Thus, novel methods to induce sustained in vivo osteogenesis should improve the pathophysiology of the disease. Our earlier studies established long-term efficacy of rAAV-transduced mesenchymal stem cells (MSC) to selectively engraft to bone, repopulate and express a transgene in a mouse model. Preliminary studies pertaining to this application indicated that recombinant AAV-2 (rAAV) transduces human and murine MSC and osteoprogenitors in high-efficiency and that rAAV-mediated transfer of bone morphogenetic protein-2 (BMP-2) leads to their differentiation into osteoblast lineage. Thus, we hypothesize that autologous transplantation of culture-expanded MSC, transduced with rAAV encoding BMP-2 under the control of osteoprogenitor-specific promoters, will result in osteoblast enrichment and increased bone mass. In the current proposal, we will evaluate this hypothesis to: 1) Determine the engraftment of ex vivo cultured MSC and optimize enrichment of homing to bone by ectopic expression of a bone homing signal, 2) Determine bone-specific expression of BMP-2 and its osteogenic significance in vivo and 3) Determine the effects of AAV-mediated gene therapy in osteopenic mice models in vivo. A successful outcome of these studies may form the basis for future development of gene therapy approaches for osteoporosis and other osteopenic diseases in humans.

描述（由申请人提供）：腺相关病毒（AAV）向量是长期治疗代谢缺陷的理想选择。独特的特征，包括非致病性，低免疫原性和稳定表达，从而成功地导致了该载体的临床前和临床评估。使用RAAV的基因治疗的潜力之一是代谢骨缺损，其特征在于骨质疏松症（如骨质疏松症）的骨骼质量减少或在脊柱融合和骨折中的合成代谢骨重塑的机制不足。尽管目前可用于骨质减少症的荷尔蒙和药物疗法旨在防止骨细胞造成进一步的骨骼破坏，但通过增加骨生成的事件，针对增加骨骼的疗法将非常有益。由于无效的递送方法以及药物和纯化的蛋白质的短期半衰期，合成代谢剂增加骨密度的处理受到限制。因此，诱导持续体内成骨的新方法应改善该疾病的病理生理。我们的早期研究确定了RAAV转导的间充质干细胞（MSC）的长期疗效，以选择性地植入骨骼，重新填充并在小鼠模型中表达转基因。与该应用有关的初步研究表明，重组AAV-2（RAAV）在高效效率方面转导人和鼠MSC以及骨基因生成剂，并且RAAV介导的骨形态发生蛋白-2（BMP-2）的转移导致其分化为骨细胞系。因此，我们假设用RAAV编码BMP-2在骨基因生激素特异性启动子的控制下对培养膨胀的MSC进行自体移植，将导致成骨细胞富集和增加的骨骼质量。在当前的建议中，我们将评估以下假设：1）确定过体培养的MSC的植入，并通过异位表达骨骼信号的异位表达来优化对骨骼的富集，2）确定BMP-2的骨特异性表达及其在Vivo中的成肌意义的骨骼特异性表达，并确定Vivo中的肌动态意义，并确定AAV介导的基因疗法的效果。这些研究的成功结果可能是人类基因治疗方法未来开发基因治疗方法的基础。