Gene therapy methods in bone and joint disorders
Gene therapy methods in bone and joint disorders
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DOI：
10.1080/17453690610046512
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发表时间：
2007-01
影响因子：
3.7
通讯作者：
M. Ulrich‐Vinther
中科院分区：
文献类型：
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作者：
M. Ulrich‐Vinther
基因治疗是一种将新基因引入细胞以通过恢复或增加基因表达来治疗疾病的技术。已经鉴定了许多具有促进运动系统中组织再生能力的生长因子和其他蛋白质，但是它们的临床应用经常受到递送问题的阻碍。原则上，这些问题可以通过递送相关基因来克服，因为治疗物质由此可以由疾病部位的局部细胞直接持续产生。关节软骨的愈合关节软骨细胞接受使用各种基因递送方法的转导。在遗传修饰后，它们能够以生物学相关水平持续表达转基因产物。我们的研究已经证明，AAV载体是一种有效的工具，基因传递到关节软骨细胞在体外以及在体内。为此，我们已经证明了AAV载体介导的TGF β 1过表达刺激软骨愈合。磨损碎屑诱导的骨质溶解：RANKL系统可能是治疗假体周围无菌性松动的关键治疗靶点。我们研究了使用AAV载体的OPG基因转移是否对骨科磨损碎片诱导的骨丢失具有保护作用。在破骨细胞生成和骨片吸收测定中，通过消耗破骨细胞生成和减少骨吸收证明了转基因OPG的生物活性。使用体内碎片诱导的骨吸收模型，我们证明了在接受AAV-OPG基因治疗的动物中完全抑制骨质溶解。骨折愈合与骨质疏松症未来OPG治疗骨质疏松症的成功高度依赖于其对骨折愈合和重塑的影响。使用体内骨折愈合模型，我们的研究表明，AAV-OPG基因治疗与正常骨愈合不冲突，与OPG的高剂量静脉内治疗相反。然而，AAV-OPG治疗抑制了骨折线处真正皮质骨的重塑和整合。结构性同种异体骨的愈合由于缺乏成骨和重塑能力，结构性同种异体骨移植物经常发生断裂。为了克服这些局限性，我们利用了用介导体内基因转移的AAV-caALK 2载体包被的同种异体移植物。我们表明，AAV载体能够转导骨折骨痂中的邻近炎性细胞和成骨细胞，并且通过AAV-caALK 2涂层递送的BMP信号直接在同种异体移植物的皮质表面上诱导骨形成。结论本研究可被视为复杂骨科疾病基因治疗选择发展的初步步骤。因此，我们的研究代表了以下原理证明：rAAV载体促进体外有效基因转移至具有矫形相关性的一系列细胞，并且在手术时用rAAV载体的单次施用体内靶向体细胞组织可能足以长期表达治疗性蛋白质。通过rAAV载体的转基因递送的未来成功的关键是不存在对载体或基因产物的免疫应答。此外，具有调控基因表达的rAAV载体的开发需要在未来的研究中进一步关注。
BACKGROUND Gene therapy is a technique that draws on the introduction of new genes into cells for the purpose of treating disease by restoring or adding gene expression. Numerous growth factors and other proteins with the ability to promote the regeneration of tissues in the locomotive system have been identified, but their clinical use is often hindered by delivery problems. In principle, these problems can be overcome by delivering the relevant genes, as the therapeutic substances thereby can be persistently produced directly by local cells at the site of diseases. HEALING OF ARTICULAR CARTILAGE Articular chondrocytes are receptive to transduction using various gene delivery methods. Following genetic modification, they are capable of sustained expression of transgene products at biologically relevant levels. Our research has proved the AAV vector to be an effective tool for gene delivery to articular chondrocytes in vitro as well as in vivo. To this end, we have demonstrated that the AAV vector mediated TGFbeta1-overexpression stimulates cartilage anabolism. WEAR DEBRIS-INDUCED OSTEOLYSIS: The RANKL system may be a key therapeutic target in treatment of aseptic periprosthetic loosening. We investigated whether gene transfer of OPG using an AAV vector has protective effects against orthopaedic wear debris-induced bone loss. In osteoclastogenesis and in bone wafer resorption assays, the bioactivity of the transgene OPG was proven by depletion of osteoclastogenesis and reduced bone resorption. Using an in vivo model of debris-induced bone resorption, we demonstrated complete inhibition of osteolysis in animals receiving AAV-OPG gene therapy. FRACTURE HEALING IN RELATION TO OSTEOPOROSIS The success of future OPG treatment of osteoporosis is highly dependent on its effects on fracture healing and remodelling. Using an in vivo fracture healing model, our studies demonstrated that AAV-OPG gene therapy did not conflict with normal bone healing, in contrast to high-dosage intravenous treatment with OPG. However, AAV-OPG therapy depressed remodelling and integration of the genuine cortical bone at the fracture line. STRUCTURAL BONE ALLOGRAFT HEALING Structural bone allografts often fracture due to their lack of osteogenic and remodelling potiential. To overcome these limitations, we utilized allografts coated with AAV-caALK2 vector that mediated in vivo gene transfer. We showed that the AAV vector was capable of transducing adjacent inflammatory cells and osteoblasts in the fracture callus and that BMP signals delivered via AAV-caALK2 coating induced bone formation directly on the cortical surface of the allograft. CONCLUSION The presented research may be seen as initial steps towards development of gene therapeutic treatment options for complex orthopaedic diseases. As such, our studies represent proof-of-principle that the rAAV vector promotes efficient gene transfer in vitro to a spectrum of cells with orthopaedic relevance, and that in vivo targeting of somatic tissue with a single administration of a rAAV vector at the time of surgery could be sufficient for long-term expression of therapeutic proteins. Essential to the future success of transgene delivery by rAAV vectors is the absence of an immune response to either the vector or the gene product. Furthermore, development of rAAV vectors with regulatory gene expression needs further attention in future research.