Targeted expression of transforming growth factor-beta 1 in intracardiac grafts promotes vascular endothelial cell DNA synthesis.
Targeted expression of transforming growth factor-beta 1 in intracardiac grafts promotes vascular endothelial cell DNA synthesis.
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DOI：
10.1172/jci117627
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
Gou Young Koh;Seong-Jin Kim;M. Klug;K. Park;K. Park;M. Soonpaa;Loren J. Field
中科院分区：
文献类型：
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作者：
Gou Young Koh;Seong-Jin Kim;M. Klug;K. Park;K. Park;M. Soonpaa;Loren J. Field
评估了由基因修饰的骨骼肌成肌细胞组成的心内移植物长期向心脏递送重组转化生长因子-β（TGF-β）的能力。C2 C12成肌细胞用包含与修饰的TGF-β 1 cDNA融合的诱导型金属硫蛋白启动子的构建体稳定转染。当在补充有硫酸锌的培养基中培养时，携带该转基因的细胞组成性地分泌活性TGF-β 1。这些遗传修饰的成肌细胞用于在同基因C3 Heb/FeJ宿主中产生心内移植物。在植入后长达三个月观察到存活的移植物，并且对维持补充有硫酸锌的水的小鼠的免疫组织学分析揭示了稳定表达TGF-β 1的移植细胞的存在。在与表达TGF-β 1的移植物相邻的心肌中观察到明显的新生血管形成区域，如氚化胸苷掺入血管内皮细胞所证明的。血管内皮细胞DNA合成的程度可以通过改变膳食锌来调节。在移植物由未转染细胞组成的小鼠中，未观察到对血管内皮细胞的类似作用。这项研究表明，基因修饰的骨骼肌成肌细胞移植物可用于影响重组分子向心脏的局部、长期递送。
Intracardiac grafts comprised of genetically modified skeletal myoblasts were assessed for their ability to effect long-term delivery of recombinant transforming growth factor-beta (TGF-beta) to the heart. C2C12 myoblasts were stably transfected with a construct comprised of an inducible metallothionein promoter fused to a modified TGF-beta 1 cDNA. When cultured in medium supplemented with zinc sulfate, cells carrying this transgene constitutively secrete active TGF-beta 1. These genetically modified myoblasts were used to produce intracardiac grafts in syngeneic C3Heb/FeJ hosts. Viable grafts were observed as long as three months after implantation, and immunohistological analyses of mice maintained on water supplemented with zinc sulfate revealed the presence of grafted cells which stably expressed TGF-beta 1. Regions of apparent neovascularization, as evidenced by tritiated thymidine incorporation into vascular endothelial cells, were observed in the myocardium which bordered grafts expressing TGF-beta 1. The extent of vascular endothelial cell DNA synthesis could be modulated by altering dietary zinc. Similar effects on the vascular endothelial cells were not seen in mice with grafts comprised of nontransfected cells. This study indicates that genetically modified skeletal myoblast grafts can be used to effect the local, long-term delivery of recombinant molecules to the heart.