Efficient bone formation by gene transfer of human LIM mineralization protein-3
Efficient bone formation by gene transfer of human LIM mineralization protein-3
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DOI：
10.1038/sj.gt.3302207
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发表时间：
2004-04-01
期刊：
影响因子：
5.1
通讯作者：
Robbins, P
中科院分区：
文献类型：
--
作者：
Pola, E;Gao, W;Robbins, P
LIM 矿化蛋白（LMP）是成骨细胞分化程序的新型正调节因子。在人类中，已鉴定出三种不同的 LMP 剪接变体：LMP-1、LMP-2 和 LMP-3。人LMP-1 (hLMP-1)的基因转移诱导参与骨形成的基因的表达，包括某些骨形态发生蛋白(BMP)，促进体外骨结节形成、体内异位骨形成，并且在后胸椎和腰椎融合的动物模型中具有治疗作用。为了检查 LMP-3 体外和体内的骨诱导特性，我们生成了表达密码子优化的 hLMP-3 的质粒和腺病毒载体。在这里，我们证明 hLMP-3 的基因转移诱导骨特异性基因骨钙蛋白、骨桥蛋白和骨唾液蛋白的表达，并诱导前成骨细胞和成纤维细胞中的骨矿化。我们还证明，hLMP-3 能够以剂量依赖性方式诱导人间充质干细胞中的骨矿化和骨特异性基因、BMP-2、OSX、RunX2 和碱性磷酸酶的表达。最后，我们证明将 hLMP-3 直接基因转移到小鼠骨骼肌中比 BMP-2 更有效地导致异位骨形成。这些结果证明hLMP-3基因转移可用于促进细胞培养物和体内的骨形成，其效果与BMP-2一样或更有效，从而确立了直接基因递送hLMP-3以在体内产生骨的可行性和功效。这些结果表明 hLMP-3 的基因转移可以开发为临床应用的骨诱导治疗剂。
LIM mineralization protein (LMP) is a novel positive regulator of the osteoblast differentiation program. In humans, three different LMP splice variants have been identified: LMP-1, LMP-2, and LMP-3. Gene transfer of human LMP-1 (hLMP-1) induces expression of genes involved in bone formation, including certain bone morphogenetic proteins (BMPs), promotes bone nodule formation in vitro, ectopic bone formation in vivo, and is therapeutic in animal models of posterior thoracic and lumbar spine fusion. To examine the osteoinductive properties of the LMP-3 in vitro and in vivo, we have generated plasmid and adenoviral vectors expressing codon-optimized hLMP-3. Here we demonstrate that gene transfer of hLMP-3 induces expression of the bone-specific genes osteocalcin, osteopontin, and bone sialoprotein and induced bone mineralization in preosteoblastic and fibroblastic cells. We also demonstrate that hLMP-3 is able to induce bone mineralization and the expression of the bone-specific genes, BMP-2, OSX, RunX2, and alkaline phosphatase in human mesenchymal stem cells in a dose-dependent manner. Finally, we demonstrate that direct gene transfer of hLMP-3 into murine skeletal muscle results in ectopic bone formation more efficiently than BMP-2. These results demonstrate that hLMP-3 gene transfer can be used to promote bone formation in cell culture and in vivo as or more efficiently than BMP-2, thus establishing feasibility and efficacy of direct gene delivery of hLMP-3 to produce bone in vivo. These results suggest that gene transfer of hLMP-3 could be developed as a bone-inductive therapeutic agent for clinical applications.