A novel library screen identifies immunosuppressors that promote osteoblast differentiation.
A novel library screen identifies immunosuppressors that promote osteoblast differentiation.
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DOI：
10.1016/j.bone.2012.03.001
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发表时间：
2012-06
期刊：
影响因子：
4.1
通讯作者：
Montano, Monty
中科院分区：
文献类型：
--
作者：
Darcy, Ariana;Meltzer, Micah;Miller, Joseph;Lee, Steven;Chappell, Scott;Donck, Kris Ver;Montano, Monty
破骨细胞介导的骨吸收增加和/或成骨细胞介导的骨沉积减少可损害骨稳态。虽然许多努力都集中在治疗破骨细胞吸收，有较少的重点，确定促进成骨细胞功能的策略。在此，我们描述了一种高通量筛选试验，以选择增加骨形态发生蛋白-2（BMP-2）介导的成骨细胞谱系定型的小分子。在对5405种化合物进行初步筛选后，包括FDA批准的药物、已知的生物活性物质和具有新型化学成分的化合物，我们确定了45种促进成骨细胞定向的小分子。在45种候选药物中，有一系列广泛的类别，包括9种类维生素A类似物/衍生物和4种免疫抑制剂，特别是雷帕霉素和FK-506，它们被选择用于进一步研究。用雷帕霉素或FK-506单独或与BMP-2协同处理成骨细胞前体细胞，增加了磷酸化Smad 1/5/8蛋白的水平和Runx-2、Osx和Smad-7的转录，与促进成骨细胞分化的作用一致。只有FK-506能够增强骨钙素转录本和茜素红染色，这两种分化的晚期标志物。当外源性TGF-β1处理抑制成骨细胞分化时，雷帕霉素（而不是FK-506）能够拯救分化标志物的表达，表明这些化合物具有不同但重叠的活性。总的来说，这些数据增加了对成骨细胞生成途径的理解，支持非冗余免疫抑制剂信号传导的作用，并为发现影响骨重建的潜在治疗化合物提供了一种新方法。
Bone homeostasis can be compromised by an increase in osteoclast-mediated resorption and/or a decrease in osteoblast-mediated bone deposition. While many efforts have focused on treating osteoclast resorption, there has been less emphasis on identifying strategies for promoting osteoblast function. Herein, we describe a high-throughput screening assay to select for small molecules that augment bone morphogenetic protein-2 (BMP-2)-mediated osteoblast lineage commitment. After an initial screen of 5405 compounds; consisting of FDA-approved drugs, known bioactives, and compounds with novel chemical makeup, we identified 45 small molecules that promoted osteoblast commitment. Of the 45 candidates, there was a broad array of classes that included nine retinoid analogs/derivatives and four immunosuppressants, notably rapamycin and FK-506, which were chosen for further study. Treatment of osteoblast precursor cells with rapamycin or FK-506, either alone, or synergistically with BMP-2, increased levels of phospho-Smad1/5/8 protein and transcription of Runx-2, Osx and Smad-7, consistent with a role in promoting osteoblast differentiation. Only FK-506 was able to enhance osteocalcin transcripts and alizarin red staining, both late markers for differentiation. When osteoblast differentiation was suppressed with exogenous TGF-β1 treatment, rapamycin (but not FK-506) was able to rescue expression of differentiation markers, indicating distinct but overlapping activity of these compounds. Collectively, these data add to an understanding of pathways engaged in osteoblastogenesis, support a role for non-redundant immunosuppressant signaling, and provide a novel approach for the discovery of potentially therapeutic compounds that affect bone remodeling.
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影响因子：
4.1
作者：
Choo, Min-Kyung;Yeo, Hyeonju;Zayzafoon, Majd
通讯作者：
Zayzafoon, Majd
影响因子：
3.7
作者：
Astrinidis, Aristotelis;Kim, Jiyoon;Azizkhan-Clifford, Jane
通讯作者：
Azizkhan-Clifford, Jane
影响因子：
--
作者：
Hamawy, MM
通讯作者：
Hamawy, MM
影响因子：
17.1
作者：
Cao, Kan;Graziotto, John J.;Collins, Francis S.
通讯作者：
Collins, Francis S.
影响因子：
--
作者：
Hoemann, C. D.;El-Gabalawy, H.;McKee, M. D.
通讯作者：
McKee, M. D.