Transfer of proalpha2(I) cDNA into cells of a murine model of human Osteogenesis Imperfecta restores synthesis of type I collagen comprised of alpha1(I) and alpha2(I) heterotrimers in vitro and in vivo.
Transfer of proalpha2(I) cDNA into cells of a murine model of human Osteogenesis Imperfecta restores synthesis of type I collagen comprised of alpha1(I) and alpha2(I) heterotrimers in vitro and in vivo.
复制标题
将 proalpha2(I) cDNA 转移到人成骨不全小鼠模型的细胞中，可在体外和体内恢复由 alpha1(I) 和 alpha2(I) 异三聚体组成的 I 型胶原蛋白的合成。
DOI：
10.1002/jcb.1209
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发表时间：
2001
影响因子：
4
通讯作者：
Robbins,P
中科院分区：
文献类型：
--
作者：
Niyibizi,C;Smith,P;Mi,Z;Phillips,CL;Robbins,P
Theoimmouse是人成骨不全（OI）的模型，其具有proα2（I）链的合成缺陷。从小鼠分离的细胞合成α1（I）胶原同源三聚体，并在组织中积累。为探讨基因治疗骨不连的可行性，将小鼠前α2（I）cDNA插入腺病毒载体中，转染小鼠骨髓基质细胞。在巨细胞病毒早期启动子控制下的鼠cDNA由转导的细胞表达。对转导细胞合成的胶原蛋白的分析表明，细胞合成稳定的I型胶原蛋白，其由2：1正确比例的α1（I）和α2（I）异源三聚体组成。当用重组人骨形态发生蛋白2处理转导的细胞时，胶原蛋白被有效地分泌，并且细胞保留成骨潜能，如碱性磷酸酶活性的表达所示。将携带鼠前α2（I）cDNA的病毒注射到皮肤中，证明在注射部位合成由α1和α2链组成的I型胶原。这些初步数据表明，胶原基因可以转移到骨髓基质细胞以及成纤维细胞在体内，基因有效地表达。这些数据鼓励进一步研究某些形式的OI的基因替代和使用骨髓基质细胞作为载体将治疗基因传递到骨。© 2001 Wiley利斯公司
Theoimmouse is a model of human Osteogenesis Imperfecta (OI) that has deficient synthesis of proα2(I) chains. Cells isolated fromoimmice synthesize α1(I) collagen homotrimers that accumulate in tissues. To explore the feasibility of gene therapy for OI, a murine proα2(I) cDNA was inserted into an adenovirus vector and transferred into bone marrow stromal cells isolated fromoimmice femurs. The murine cDNA under the control of the cytomegalovirus early promoter was expressed by the transduced cells. Analysis of the collagens synthesized by the transduced cells demonstrated that the cells synthesized stable type I collagen comprised of α1(I) and α2(I) heterotrimers in the correct ratio of 2:1. The collagen was efficiently secreted and also the cells retained the osteogenic potential as indicated by the expression of alkaline phosphatase activity when the transduced cells were treated with recombinant human bone morphogenetic protein 2. Injection of the virus carrying the murine proα2(I) cDNA intooimskin demonstrated synthesis of type I collagen comprised of α1 and α2 chains at the injection site. These preliminary data demonstrate that collagen genes can be transferred into bone marrow stromal cells as well as fibroblasts in vivo and that the genes are efficiently expressed. These data encourage further studies in gene replacement for some forms of OI and use of bone marrow stromal cells as vehicles to deliver therapeutic genes to bone. © 2001 Wiley‐Liss, Inc.