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Bone Induction by Human Bone Morphogenetic Protein Expressing Adenoviral Vector

表达腺病毒载体的人骨形态发生蛋白的骨诱导

基本信息

批准号：
12307048
负责人：
IIZUKA Tadahiko
金额：
$ 27.49万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

Large quantity of bone morphogenetic protein (BMP) is required with primates to get the bone induction for which to become stable when BMP is used under the condition of the protein. This is a big problem for clinical application. There is a possibility to solve this problem by using the BMP gene transfer method with an adenoviral vector because the infection efficiency of the adenovirus is especially high with primates.We constructed a human bone morphogenetic protein-2 (BMP-2)-expressing adenoviral vector. C2C12 cells were infected in vitro with the vital vector. An efficient gene transfer to the C2C12 and BMP-2 expression in the cells infected with the viral vector were confirmed. C2C12 cells transfected with the BMP-2 gene by the vector showed intracellular accumulation alkaline phosphatase, and also produced and secreted osteocalcim into the culture medium, demonstrating that in vitro transfer of the BMP-2 gene to these cells had caused the transformation of myoblasts to the osteo … More blast lineage. Then we examined the effectiveness of BMP-2 gene transfer by the viral vector in vivo. Osteoinduction was seen in muscle when the viral vector was injected immunosuppression. We also evaluated the time course of osteoinduction by the viral vector in immunocompetent and immunosuppressed rats. In vivo bone formation at 14 and 21 days after injection was only seen in the immunosuppressed rats, while most of the cells infected with the viral vector were eliminated by the host immune system soon after injection in the immunocompetent rats. Therefore, we evaluated osteoinduction by the viral vector using biomaterial (atelopeptide collagen) to mask the immune response or local application of immunosuppressant in immunocompetent rats.Our results suggested that gene therapy with the BMP-2-expressing adenoviral vector could be performed under transient immunosuppression, with local application of immunosuppressant, or with effective masking of the host immune response and may be useful for bone reconstruction. Less

灵长类动物需要大量的骨形态发生蛋白（BMP）才能获得骨诱导，在该蛋白的条件下使用BMP才能使骨诱导变得稳定。这对于临床应用来说是一个很大的问题。由于腺病毒对灵长类动物的感染效率特别高，因此，利用腺病毒载体进行BMP基因转移有可能解决这个问题。用活载体体外感染C2 C12细胞。证实了有效的基因转移到C2 C12和BMP-2在用病毒载体感染的细胞中的表达。通过载体转染BMP-2基因的C2 C12细胞显示细胞内碱性磷酸酶的积累，并且还产生并分泌骨钙素到培养基中，这表明在体外将BMP-2基因转移到这些细胞中引起成肌细胞向成骨细胞的转化。 ...更多信息胚系然后，我们在体内检测了BMP-2基因通过病毒载体转移的有效性。当注射免疫抑制剂时，在肌肉中观察到骨诱导。我们还评估了免疫活性和免疫抑制大鼠中病毒载体诱导骨形成的时间过程。注射后14天和21天的体内骨形成仅见于免疫抑制大鼠，而免疫活性大鼠注射后不久，大部分病毒载体感染的细胞被宿主免疫系统清除。因此，我们使用生物材料评价了病毒载体的骨诱导作用我们的研究结果表明，用BMP-2表达腺病毒载体的基因治疗可以在短暂的免疫抑制下进行，局部应用免疫抑制剂，或者有效地掩蔽宿主免疫应答，并且可用于骨重建。少