Regulation of Osteoclast Function : Role of the Cbl-PI3K Complex

破骨细胞功能的调节：Cbl-PI3K 复合物的作用

基本信息

批准号：
8270388
负责人：
Archana Sanjay
金额：
$ 32.2万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8270388
关键词：
1-Phosphatidylinositol 3-Kinase Actins Adaptor Signaling Protein Address Affect American Applications Grants Arthritis Binding Binding Sites Biochemical Biology Bone Resorption Bone remodeling C-terminal Cell physiology Cells Collagen Complex Cultured Cells Cyclophosphamide Cytoskeleton Data Defect Development Event Family Genes Guanine Guanine Nucleotide Exchange Factors Guanosine Triphosphate Phosphohydrolases Hand Health Care Costs In Vitro Integrins Knock-in Mouse Knock-out Link Lipids Maps Measurement Mediating Metastatic Neoplasm to the Bone Methodology Molecular Monomeric GTP-Binding Proteins Mus Mutate Osteoclasts Osteoporosis Pathology Periodontal Diseases Phosphorylation Phosphotransferases Play Proteins Recruitment Activity Regulation Research Research Personnel Role Serum Signal Pathway Signal Transduction Signaling Protein Site Stimulus Structure Structure-Activity Relationship Therapeutic Intervention Tyrosine Tyrosine Phosphorylation Work base bone bone loss cell motility design gain of function in vivo loss of function migration mouse model mutant novel therapeutic intervention protein protein interaction response rho skeletal tissue src-Family Kinases ubiquitin-protein ligase

项目摘要

Description (provided by applicant): Osteoclasts are the principal cells that migrate to the potential bone resorption sites and resorb bone under normal and pathological conditions. Our preliminary data has shown that in osteoclasts, deletion of cbl gene impairs osteoclast migration during development. Similarly, the ability of Cbl-/- osteoclasts to reorganize the actin cytoskeleton in response to chemotactic stimuli is also significantly compromised. We have also shown that tyrosine phosphorylation-dependent binding of Cbl to PI3-Kinase, a signaling protein required for osteoclast function, is one of the key requirements to organize the signaling events upon integrin activation. Abolishing the interaction between Cbl and PI3K kinase significantly compromises the bone resorbing capacity of osteoclasts both in vivo and in vitro, indicating that formation of the Cbl-PI3K complex and the subsequent signaling events downstream of this complex are necessary for proper functioning of osteoclasts. Although some progress has been made in understanding the roles of Cbl and PI3K in osteoclast biology the questions that remain to be answered are: what is the mechanism of interaction between the two proteins and what are the downstream events that link this complex to osteoclast function? We have addressed these issues the following specific aims: AIM-1. Determines which tyrosine kinase (Src or Syk) is critical for phosphorylation of the CblY731 downstream of av¿3 activation and the effect of modulating the phosphorylation of CblY731 on osteoclast migration and bone resorption in vitro. AIM-2. Determines the in vivo impact of Cbl-PI3K complex on skeletal tissue in general and osteoclast function particular by using two different knock-in mouse models in which the Cbl-PI3K interaction is either abolished or constitutively activated. AIM-3. Examines the signaling events downstream of Cbl-PI3K interaction that are important in regulating actin cytoskeleton. The proposed studies will unequivocally demonstrate the interdependence of the events in osteoclast downstream of av¿3-mediated signaling. The specific aims of the proposal will be accomplished through the application of varieties of approaches and methodologies. This includes in vivo studies with knock-out and knock-in mice, biochemical analysis of cultured cells expressing wild type and mutant proteins and biochemical studies of cells isolated form the genetically modified mice. Successful completion of these aims will demonstrate the positive role played by Cbl in osteoclast function and will increase our understanding of the regulation of bone resorption, potentially identifying novel therapeutic intervention to control bone loss related to osteoporosis arthritis, bone metastasis and periodontal disease. Osteoporosis and other bone-related maladies afflict 150 million Americans and constitute a significant burden to ever-rising health care cost. The research proposed here will help in understanding the basic biology behind bone resorption and will help identifying targets for therapeutic interventions to control bone loss.

描述（由申请人提供）：破骨细胞是在正常和病理条件下迁移到潜在骨吸收位点并吸收骨的主要细胞，我们的初步数据表明，在破骨细胞中，cbl基因的缺失会损害破骨细胞在发育过程中的迁移。 Cbl-/-破骨细胞响应趋化刺激而重组肌动蛋白细胞骨架的能力也显着受损。我们还表明酪氨酸磷酸化依赖性结合。 Cbl 与 PI3-激酶（破骨细胞功能所需的信号蛋白）的结合是在整合素激活时组织信号传导事件的关键要求之一。消除 Cbl 和 PI3K 激酶之间的相互作用会显着损害破骨细胞的骨吸收能力。体外研究表明，Cbl-PI3K 复合物的形成以及该复合物下游的信号传导事件对于破骨细胞的正常功能是必要的，尽管在理解其作用方面已经取得了一些进展。 Cbl 和 PI3K 在破骨细胞生物学中的作用尚待回答的问题是：这两种蛋白质之间的相互作用机制是什么？将这种复合物与破骨细胞功能联系起来的下游事件是什么？我们已经解决了这些问题，具体目标如下： AIM-1。确定哪种酪氨酸激酶（Src 或 Syk）对于 av¿ 的 CblY731 下游的磷酸化至关重要。 3 体外 AIM-2 的激活和调节 CblY731 磷酸化对破骨细胞迁移和骨吸收的影响通过使用两种不同的敲入，确定 Cbl-PI3K 复合物对骨骼组织和破骨细胞功能的体内影响。 Cbl-PI3K 相互作用被废除或组成型激活的小鼠模型检查 Cbl-PI3K 相互作用下游的信号事件。这些对于调节肌动蛋白细胞骨架很重要，拟议的研究将明确证明破骨细胞下游事件的相互依赖性。 3 介导的信号传导。该提案的具体目标将通过应用各种方法和方法来实现，其中包括对敲除和敲入小鼠进行体内研究，对表达野生型和突变蛋白的培养细胞进行生化分析。对从转基因小鼠中分离出的细胞进行的生物化学研究的成功完成将证明 Cbl 在破骨细胞功能中发挥的积极作用，并将增加我们对骨吸收调节的理解，有可能确定控制骨丢失相关的新治疗干预措施。对于骨质疏松性关节炎，骨转移和牙周病以及其他与骨相关的疾病困扰着 1.5 亿美国人，并给不断上涨的医疗保健费用带来了沉重负担。本文提出的研究将有助于了解骨吸收背后的基本生物学，并有助于确定骨吸收的目标。控制骨质流失的治疗干预措施。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Loss of Cbl-PI3K interaction in mice prevents significant bone loss following ovariectomy.

DOI：
10.1016/j.bone.2014.06.013
发表时间：
2014-10
期刊：
BONE
影响因子：
4.1
作者：
Adapala, Naga Suresh;Holland, Danielle;Scanlon, Vanessa;Barbe, Mary F.;Langdon, Wallace Y.;Tsygankov, Alexander Y.;Lorenzo, Joseph A.;Sanjay, Archana
通讯作者：
Sanjay, Archana

Cbl-PI3K interaction regulates Cathepsin K secretion in osteoclasts.

DOI：
10.1016/j.bone.2019.07.009
发表时间：
2019-10
期刊：
Bone
影响因子：
4.1
作者：
Jungeun Yu;N. Adapala;Laura Doherty;A. Sanjay
通讯作者：
Jungeun Yu;N. Adapala;Laura Doherty;A. Sanjay

Role of Cbl-PI3K Interaction during Skeletal Remodeling in a Murine Model of Bone Repair.

Cbl-PI3K 相互作用在小鼠骨修复模型骨骼重塑过程中的作用。

DOI：
10.1371/journal.pone.0138194
发表时间：
2015
期刊：
PloS one
影响因子：
3.7
作者：
Scanlon,Vanessa;Soung,DoYu;Adapala,NagaSuresh;Morgan,Elise;Hansen,MarcF;Drissi,Hicham;Sanjay,Archana
通讯作者：
Sanjay,Archana

Abrogation of Cbl-PI3K interaction increases bone formation and osteoblast proliferation.

DOI：
10.1007/s00223-011-9531-z
发表时间：
2011-11
期刊：
CALCIFIED TISSUE INTERNATIONAL
影响因子：
4.2
作者：
Brennan, Tracy;Adapala, Naga Suresh;Barbe, Mary F.;Yingling, Vanessa;Sanjay, Archana
通讯作者：
Sanjay, Archana

Loss of Cbl-PI3K interaction enhances osteoclast survival due to p21-Ras mediated PI3K activation independent of Cbl-b.