High-throughput screening for modulators of ACVR1 transcription: discovery of potential therapeutics for fibrodysplasia ossificans progressiva.
High-throughput screening for modulators of ACVR1 transcription: discovery of potential therapeutics for fibrodysplasia ossificans progressiva.
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DOI：
10.1242/dmm.023929
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发表时间：
2016-06-01
影响因子：
4.3
通讯作者：
Bocciardi R
中科院分区：
文献类型：
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作者：
Cappato S;Tonachini L;Giacopelli F;Tirone M;Galietta LJ;Sormani M;Giovenzana A;Spinelli AE;Canciani B;Brunelli S;Ravazzolo R;Bocciardi R
ACVR1基因编码骨形态发生蛋白（BMP）的I型受体。ACVR1中的激活突变导致进行性骨化性纤维发育不良（FOP），这是一种罕见疾病，其特征为先天性脚趾畸形和进行性异位软骨内骨化，导致严重和累积残疾。到目前为止，还没有治疗方法可以防止软组织肿胀（突发）触发骨化过程。为了寻找FOP的新治疗策略，我们开发了一种高通量筛选（HTS）试验，以在已批准用于治疗其他疾病的药物中鉴定ACVR1基因表达的抑制剂。基于ACVR1启动子测定的筛选之后进行体外和体内测试以验证和表征候选分子。在调节ACVR 1启动子活性的化合物中，我们选择了一种显示出最高抑制作用的化合物，双嘧达莫，一种目前用作血小板抗聚集剂的药物。通过体外细胞测定，可检测到对ACVR1基因表达、对整个Smad依赖性BMP信号传导途径以及对软骨和成骨分化过程的抑制作用。此外，潘生丁减少体内异位骨形成的过程。我们的药物重新定位策略已导致确定双嘧达莫作为治疗FOP的可能治疗工具。此外，我们的研究还确定了一个管道的测定，将是有用的评价其他药物抑制剂的异位骨化。总结：我们描述了通过一系列体外和体内试验筛选和验证FDA批准的化合物，将双嘧达莫作为FOP的潜在治疗工具进行鉴定。
The ACVR1 gene encodes a type I receptor of bone morphogenetic proteins (BMPs). Activating mutations in ACVR1 are responsible for fibrodysplasia ossificans progressiva (FOP), a rare disease characterized by congenital toe malformation and progressive heterotopic endochondral ossification leading to severe and cumulative disability. Until now, no therapy has been available to prevent soft-tissue swelling (flare-ups) that trigger the ossification process. With the aim of finding a new therapeutic strategy for FOP, we developed a high-throughput screening (HTS) assay to identify inhibitors of ACVR1 gene expression among drugs already approved for the therapy of other diseases. The screening, based on an ACVR1 promoter assay, was followed by an in vitro and in vivo test to validate and characterize candidate molecules. Among compounds that modulate the ACVR1 promoter activity, we selected the one showing the highest inhibitory effect, dipyridamole, a drug that is currently used as a platelet anti-aggregant. The inhibitory effect was detectable on ACVR1 gene expression, on the whole Smad-dependent BMP signaling pathway, and on chondrogenic and osteogenic differentiation processes by in vitro cellular assays. Moreover, dipyridamole reduced the process of heterotopic bone formation in vivo. Our drug repositioning strategy has led to the identification of dipyridamole as a possible therapeutic tool for the treatment of FOP. Furthermore, our study has also defined a pipeline of assays that will be useful for the evaluation of other pharmacological inhibitors of heterotopic ossification. Summary: We describe the identification of dipyridamole as a potential therapeutic tool for FOP, through a series of in vitro and in vivo assays to screen and validate FDA-approved compounds.