Pharmacological inhibition of PHOSPHO1 suppresses vascular smooth muscle cell calcification.
Pharmacological inhibition of PHOSPHO1 suppresses vascular smooth muscle cell calcification.
复制标题
DOI：
10.1002/jbmr.1733
复制
发表时间：
2013-01
影响因子：
6.2
通讯作者：
Millan, Jose Luis
中科院分区：
文献类型：
--
作者：
Kiffer-Moreira, Tina;Yadav, Manisha C.;Zhu, Dongxing;Narisawa, Sonoko;Sheen, Campbell;Stec, Boguslaw;Cosford, Nicholas D.;Dahl, Russell;Farquharson, Colin;Hoylaerts, Marc F.;MacRae, Vicky E.;Millan, Jose Luis
关键词：
中膜血管钙化（MVC）常见于慢性肾病、肥胖和衰老患者。MVC是一种类似于骨骼矿化的主动调节过程，由血管平滑肌细胞（VSMC）的软骨成骨转化引起。在这里，我们使用矿化的小鼠VSMCs研究磷酸酶1的表达，磷酸酶参与软骨内骨化过程中基质囊泡介导的矿化起始的第一步。在钙化条件下培养的野生型（WT）VSMCs表现出Phospho 1基因表达增加，Phospho 1-/- VSMCs未能在体外矿化。使用天然PHOSPHO 1底物，通过高通量筛选和机制分析鉴定了PHOSPHO 1的有效和特异性抑制剂，并选择了两种，命名为MLS-0390838和MLS-0263839，用于进一步分析。通过靶向PHOSPHO 1功能，评估了它们在预防VSMC钙化方面的有效性，单独使用以及与强效组织非特异性碱性磷酸酶（TNAP）抑制剂MLS-0038949联合使用。MLS-0263839对矿化WT细胞（添加无机磷酸盐培养）中的PHOSPHO 1抑制使培养物中的钙化减少至对照的41.8% ± 2.0。MLS-0263839对PHOSPHO 1的联合抑制和MLS-0038949对TNAP的联合抑制可显著降低钙化至对照的20.9% ± 0.74。此外，双重抑制策略影响了几种矿化相关酶的表达，同时增加了平滑肌细胞标志物Acta 2的表达。我们的结论是，PHOSPHO 1起着至关重要的作用，在VSMC矿化和“磷酸酶抑制”可能是一个有用的治疗策略，以减少MVC。
Medial vascular calcification (MVC) is common in patients with chronic kidney disease, obesity, and aging. MVC is an actively regulated process that resembles skeletal mineralization, resulting from chondro-osteogenic transformation of vascular smooth muscle cells (VSMCs). Here, we used mineralizing murine VSMCs to study the expression of PHOSPHO1, a phosphatase that participates in the first step of matrix vesicles-mediated initiation of mineralization during endochondral ossification. Wild-type (WT) VSMCs cultured under calcifying conditions exhibited increased Phospho1 gene expression and Phospho1-/- VSMCs failed to mineralize in vitro. Using natural PHOSPHO1 substrates, potent and specific inhibitors of PHOSPHO1 were identified via high-throughput screening and mechanistic analysis and two, designated MLS-0390838 and MLS-0263839, were selected for further analysis. Their effectiveness in preventing VSMC calcification by targeting PHOSPHO1 function was assessed, alone and in combination with a potent tissue-nonspecific alkaline phosphatase (TNAP) inhibitor MLS-0038949. PHOSPHO1 inhibition by MLS-0263839 in mineralizing WT cells (cultured with added inorganic phosphate) reduced calcification in culture to 41.8% ± 2.0 of control. Combined inhibition of PHOSPHO1 by MLS-0263839 and TNAP by MLS-0038949 significantly reduced calcification to 20.9% ± 0.74 of control. Furthermore, the dual inhibition strategy affected the expression of several mineralization-related enzymes while increasing expression of the smooth muscle cell marker Acta2. We conclude that PHOSPHO1 plays a critical role in VSMC mineralization and that “phosphatase inhibition” may be a useful therapeutic strategy to reduce MVC.
登录
查看更多内容
DOI：
10.1359/jbmr.091023
发表时间：
2010-04
期刊：
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
影响因子：
--
作者：
Ciancaglini P;Yadav MC;Simão AM;Narisawa S;Pizauro JM;Farquharson C;Hoylaerts MF;Millán JL
通讯作者：
Millán JL
影响因子：
4.1
作者：
MacRae, Vicky E.;Davey, Megan G.;Farquharson, Colin
通讯作者：
Farquharson, Colin
影响因子：
14.9
作者：
Jaroszewski L;Li Z;Cai XH;Weber C;Godzik A
通讯作者：
Godzik A
影响因子：
3.8
作者：
Markello, Thomas C.;Pak, Laura K.;St Hilaire, Cynthia;Dorward, Heidi;Ziegler, Shira G.;Chen, Marcus Y.;Chaganti, Krishna;Nussbaum, Robert L.;Boehm, Manfred;Gahl, William A.
通讯作者：
Gahl, William A.
影响因子：
6
作者：
Harmey, D;Hessle, L;Millán, JL
通讯作者：
Millán, JL