Ex vivo gene therapy with stromal cells transduced with a retroviral vector containing the BMP4 gene completely heals critical size calvarial defect in rats
Ex vivo gene therapy with stromal cells transduced with a retroviral vector containing the BMP4 gene completely heals critical size calvarial defect in rats
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DOI：
10.1038/sj.gt.3301772
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发表时间：
2002-08-01
期刊：
影响因子：
5.1
通讯作者：
Baylink, DJ
中科院分区：
文献类型：
--
作者：
Gysin, R;Wergedal, JE;Baylink, DJ
为了开发成功的基因治疗系统来治愈骨缺损，我们开发了一种基于鼠白血病病毒（MLV）的逆转录病毒系统，该系统表达具有高转导效率的人骨形态发生蛋白（BMP）4转基因。通过将 2.5 x 10(6) 转导基质细胞嵌入明胶基质中，然后将其放置在同系大鼠颅盖的临界尺寸缺损中，测试 BMP4 转导细胞的骨形成潜力。不含细胞或含有未转导基质细胞的明胶基质是两个对照组。 4周后，实验大鼠的缺损区域完全被新骨填充，而任一对照组均出现有限的骨形成，基因治疗组缺损的骨矿物质密度（BMD）为67.8 +/- 5.7 mg/cm(2)（平均值+/- s.d.，n = 4），是缺损周围骨的对照BMD（57.2 +/- 1.5）的119 +/- 10%。毫克/厘米(2))。相比之下，植入未转导基质细胞的大鼠的 BMD 低五倍（13.8 +/- 7.4 mg/cm(2)，P < 0.001）。时程研究表明，在临界尺寸缺陷处接种 2.5 x 106 植入的 BMP4 细胞后 2-4 周，BMD 呈线性增加。总之，我们开发的基于逆转录病毒的 BMF4 基因治疗系统具有修复大型骨骼缺陷的潜力。
In order to develop a successful gene therapy system for the healing of bone defects, we developed a murine leukemia virus (MLV)-based retroviral system expressing the human bone morphogenetic protein (BMP) 4 transgene with high transduction efficiency. The bone formation potential of BMP4 transduced cells was tested by embedding 2.5 x 10(6) transduced stromal cells in a gelatin matrix that was then placed in a critical size defect in calvariae of syngenic rats. Gelatin matrix without cells or with untransduced stromal cells were the two control groups. The defect area was completely filled with new bone in experimental rats after 4 weeks, while limited bone formation occurred in either control group, Bone mineral density (BMD) of the defect in the gene therapy group was 67.8 +/- 5.7 mg/cm(2) (mean +/- s.d., n = 4), which was 119 +/- 10% of the control BMD of bone surrounding the defect (57.2 +/- 1.5 mg/cm(2)). In contrast, BMD of rats implanted with untransduced stromal cells was fivefold lower (13.8 +/- 7.4 mg/cm(2), P < 0.001). Time course studies revealed that there was a linear increase in BMD between 2-4 weeks after inoculation of the critical size defect with 2.5 x 106 implanted BMP4 cells. In conclusion, the retroviral-based BMF4 gene therapy system that we have developed has the potential for regeneration of large skeletal defects.