Regulation of Osteoclast Activity by Calcium and cGMP

钙和 cGMP 对破骨细胞活性的调节

基本信息

批准号：
7769843
负责人：
Harry C. Blair
金额：
$ 27.88万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2012-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7769843
关键词：
Actins Address Affect Antibodies Biological Assay Ca(2+)-Transporting ATPase Calcineurin Calcium Calmodulin Calpain Cell-Matrix Junction Cells Cyclic AMP Cyclic GMP Cytoskeleton DNA Sequence Rearrangement Data Dissociation Estrogens Felis catus Figs - dietary Free Radicals Human Hydrolysis ITPR1 gene Inositol Integrin beta3 Integrins Intervention Intracellular Second Messenger Label Location Macrophage Colony-Stimulating Factor Measures Mediating Mediator of activation protein Membrane Modeling Molecular Nitric Oxide Normal Cell Osteoblasts Osteoclasts Osteoporosis Pathway interactions Peptide Hydrolases Phospholipase C Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Physiologic pulse Post-Translational Protein Processing Production Protein Tyrosine Kinase Proteins Proteolysis Regulation Research Personnel Role Second Messenger Systems Signal Transduction Site Source Stimulus Stress Stretching System TNFSF11 gene Testing autocrine bone bone cell bone mass bone turnover cGMP-dependent protein kinase I cell attachment protein cell motility cytokine inhibitor/antagonist link protein mu-calpain paracrine phosphoric diester hydrolase programs protein complex receptor reconstitution response src-Family Kinases uptake vasodilator-stimulated phosphoprotein

项目摘要

DESCRIPTION (provided by applicant): Stimuli that regulate bone mass, including estrogen, modulate nitric oxide (NO) production in bone cells. NO is an important regulator of bone degradation. Our studies showed that NO regulates osteoclast motility. Response to NO is mediated by the cGMP-dependent protein kinase I (PKG I). PKG I action on proteins at the osteoclast's attachment site allow the cell to detach from bone. This is accompanied by Ca2+ release, probably via the IP3R receptor, and by intracellular proteolysis involving mu-calpain. Motility-related changes are then reversed, allowing the osteoclast to resume bone degradation in a new location. We will study this mechanism further using human bone cells as our principal test system. In Aim I we will determine how NO, cGMP, and PKG I regulate osteoclast attachment. This will include studies of rearrangement of membrane- attachment proteins in response to NO, including VASP, migfilin and the alph-v-beta3 integrin. The regulation of NO synthesis in the presence of key stimuli including cell stretch and estrogen will be characterized in osteoclasts and in osteoblasts. VASP will be studied in PKG l-deficient cells, where it may also regulate motility induced by stimuli other than NO, such as CSF-1. PKG l-deficient cells will also be studied to evaluate NO-free radical actions, which are difficult to detect in the presence of PKG I. In Aim 2, we will define Ca2+-dependent mechanisms that are critical to completing and reversing NO-induced osteoclast motility. These studies will use pharmacological inhibitors and assays for specific mediators, including Ca2+, in normal cells and in cells deficient in key pathway constituents. We will determine the source of Ca pulses that occur in response to NO and cGMP. PKG l-induced changes in attachment proteins that activate the inositol-1,4,5-trisphosphate receptor will be characterized. We will determine how the Ca2+ activated proteinase mu-calpain functions during motility. The mechanisms by which calmodulin-activated proteins including phosphodiesterase, phosphatase, and Ca2+-ATPase terminate motility will be determined. Regulation of mu-calpain by phosphorylation and cleavage will be analyzed, and proteins that are modified by mu-calpain during motility will be identified. The mechanisms defined by these studies will highlight potential targets for pharmacological intervention in osteoporosis.

描述（由申请人提供）：调节骨骼质量的刺激，包括雌激素，调节骨细胞中的一氧化氮（NO）产生。否是骨骼降解的重要调节剂。我们的研究表明，没有调节破骨细胞运动。对NO的响应是由CGMP依赖性蛋白激酶I（PKG I）介导的。 PKG I对破骨细胞附着位点的蛋白质作用使细胞从骨骼中分离出来。这可能是通过IP3R受体以及涉及MU-升性蛋白的细胞内蛋白水解的伴随。然后将与运动相关的变化逆转，从而使破骨细胞可以在新位置恢复骨骼降解。我们将使用人骨细胞作为主要测试系统进一步研究这种机制。在目的中，我将确定否，CGMP和PKG I如何调节破骨细胞附着。这将包括针对NO的膜附着蛋白重排的研究，包括VASP，MIGFILIN和ALPH-V-BETA3整合素。在包括细胞拉伸和雌激素在内的关键刺激存在下，NO合成的调节将在破骨细胞和成骨细胞中进行表征。 VASP将在PKG L缺陷型细胞中进行研究，该细胞还可以调节刺激诱导的运动性，例如CSF-1。还将研究PKG L缺陷型细胞以评估无自由基动作，在PKG I的存在下很难检测到，在AIM 2中，我们将定义Ca2+依赖性机制，这些机制对于完成和逆转无诱导的机制至关重要破骨细胞运动。这些研究将在正常细胞和缺乏关键途径成分缺乏的细胞中使用药理抑制剂和测定法，包括CA2+在内的特定介质。我们将确定响应于NO和CGMP的Ca脉冲的来源。 PKG L诱导的固定蛋白的变化将表征激活肌醇1,4,5-三磷酸受体的变化。我们将确定在运动过程中CA2+激活的蛋白酶MU-钙蛋白钙蛋白酶功能。将确定钙调蛋白激活蛋白在内的机制，包括磷酸二酯酶，磷酸酶和Ca2+-ATPase终止运动。将分析通过磷酸化和裂解来调节MU-钙蛋白酶，并确定由MU-升性蛋白在运动过程中修饰的蛋白质。这些研究定义的机制将突出骨质疏松症药理干预的潜在靶标。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

FSH-receptor isoforms and FSH-dependent gene transcription in human monocytes and osteoclasts.

DOI：
10.1016/j.bbrc.2010.02.112
发表时间：
2010-03-26
期刊：
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
影响因子：
3.1
作者：
Robinson, Lisa J.;Tourkova, Irina;Wang, Yujuan;Sharrow, Allison C.;Landau, Michael S.;Yaroslavskiy, Beatrice B.;Sun, Li;Zaidi, Mone;Blair, Harry C.
通讯作者：
Blair, Harry C.

Calcium signalling and calcium transport in bone disease.