Gene targeting of mutant COL1A2 alleles in mesenchymal stem cells from individuals with osteogenesis imperfecta
Gene targeting of mutant COL1A2 alleles in mesenchymal stem cells from individuals with osteogenesis imperfecta
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DOI：
10.1038/sj.mt.6300339
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发表时间：
2008-01-01
影响因子：
12.4
通讯作者：
Russell, David W.
中科院分区：
文献类型：
--
作者：
Chamberlain, Joel R.;Deyle, David R.;Russell, David W.
间充质干细胞（MSC）是具有分化成多种细胞类型的能力的成体细胞，包括骨、脂肪、软骨和肌肉细胞。为了在基于细胞的治疗中有效地利用自体MSC，需要精确的遗传操作来消除致病突变的影响。我们以前使用腺相关病毒（AAV）载体的目标和突变的COL1A1基因在骨髓间充质干细胞从个人与脆骨疾病，成骨不全（OI）。在这里，我们已经使用AAV载体来抑制OI MSC中的突变型COL1A2基因，从而证明可以成功靶向负责OI的两种I型胶原基因。我们整合了改进的载体设计，以最大限度地减少随机整合的后果，促进潜在抗原的去除，并避免不需要的外显子跳跃。针对突变型COL1A2等位基因的MSC产生正常的I型前胶原并形成骨，从而证明其治疗潜力。
Mesenchymal stem cells (MSCs) are adult cells with the capacity to differentiate into multiple cell types, including bone, fat, cartilage, and muscle cells. In order to effectively utilize autologous MSCs in cell-based therapies, precise genetic manipulations are required to eliminate the effects of disease-causing mutations. We previously used adeno-associated virus (AAV) vectors to target and inactivate mutant COL1A1 genes in MSCs from individuals with the brittle bone disorder, osteogenesis imperfecta (OI). Here we have used AAV vectors to inactivate mutant COL1A2 genes in OI MSCs, thereby demonstrating that both type I collagen genes responsible for OI can be successfully targeted. We incorporated improved vector designs so as to minimize the consequences of random integration, facilitate the removal of potential antigens, and avoid unwanted exon skipping. MSCs targeted at mutant COL1A2 alleles produced normal type I procollagen and formed bone, thereby demonstrating their therapeutic potential.