Transfer, analysis, and reversion of the fibrous dysplasia cellular phenotype in human skeletal progenitors.
Transfer, analysis, and reversion of the fibrous dysplasia cellular phenotype in human skeletal progenitors.
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DOI：
10.1359/jbmr.091036
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发表时间：
2010-05
期刊：
影响因子：
--
通讯作者：
Bianco P
中科院分区：
文献类型：
--
作者：
Piersanti S;Remoli C;Saggio I;Funari A;Michienzi S;Sacchetti B;Robey PG;Riminucci M;Bianco P
使用慢病毒载体将人骨骼祖细胞工程化以稳定表达R201C突变的组成型活性Gsα。长期转导的骨骼祖细胞的特征是cAMP的产生增加，表明Gsα激活突变引起的基本细胞表型的转移。与从自然FD病变中分离的骨骼祖细胞一样，转导细胞在体内移植后可以产生骨骼，但不能产生脂肪细胞或造血微环境。在体外成骨分化，指出缺乏矿物质沉积，钝化上调的骨钙素，但增强上调的其他成骨标志物，如ALP和BSP相比，控制。观察到RANKL表达的非常有效的上调，这与体内FD病变中观察到的显著破骨细胞生成相关。稳定的转导导致选定的磷酸二酯酶（PDE）亚型mRNA的显著上调，并在总PDE活性的显着增加。这预示着在用组成型活性突变的Gsα转导的骨骼祖细胞中的适应性反应。事实上，与可测量的cAMP水平一样，转导的骨骼祖细胞的分化反应受到PDE抑制或缺乏PDE的深刻影响。最后，使用编码短发夹（sh）RNA干扰序列的慢病毒载体，我们证明了突变等位基因的选择性沉默在逆转由组成型活性Gsα引起的异常cAMP产生及其对骨骼祖细胞体外分化的一些影响方面是可行和有效的。
Human skeletal progenitors were engineered to stably express R201C mutated, constitutively active Gsα using lentiviral vectors. Long-term transduced skeletal progenitors were characterized by an enhanced production of cAMP, indicating the transfer of the fundamental cellular phenotype caused by activating mutations of Gsα. Like skeletal progenitors isolated from natural FD lesions, transduced cells could generate bone, but not adipocytes or the hematopoietic microenvironment, upon in vivo transplantation. In vitro osteogenic differentiation was noted for the lack of mineral deposition, a blunted up-regulation of osteocalcin, but with enhanced up-regulation of other osteogenic markers, such as ALP and BSP compared to controls. A very potent up-regulation of RANKL expression was observed, which correlates with the pronounced osteoclastogenesis observed in FD lesions in vivo. Stable transduction resulted in a marked up-regulation of selected phosphodiesterase (PDE) isoform mRNAs, and in a prominent increase in total PDE activity. This predicts an adaptive response in skeletal progenitors transduced with constitutively active, mutated Gsα. Indeed, like measurable cAMP levels, the differentiative responses of transduced skeletal progenitors were profoundly affected by inhibition of PDEs, or lack thereof. Finally, using lentiviral vectors encoding short hairpin (sh) RNA interfering sequences, we demonstrated that selective silencing of the mutated allele is both feasible and effective in reverting the aberrant cAMP production brought about by the constitutively active Gsα, and some of its effects on in vitro differentiation of skeletal progenitors.
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影响因子：
5.8
作者：
Persani, L;Borgato, S;Spada, A
通讯作者：
Spada, A
影响因子：
6.2
作者：
Kuznetsov, Sergei A.;Cherman, Natasha;Bianco, Paolo
通讯作者：
Bianco, Paolo
影响因子：
6.2
作者：
Corsi, A;Collins, MT;Bianco, P
通讯作者：
Bianco, P
影响因子：
5.3
作者：
Petersen, Rasmus Koefoed;Madsen, Lise;Kristiansen, Karsten
通讯作者：
Kristiansen, Karsten
影响因子：
--
作者：
NEMOZ, G;SETTE, C;CONTI, M
通讯作者：
CONTI, M