Modulation of Bone Formation by SHN3

SHN3 对骨形成的调节

基本信息

批准号：
9561942
负责人：
Matthew Blake Greenblatt
金额：
$ 42.38万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-18 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9561942
关键词：
Adaptor Signaling Protein Address Adoption Affect Age Alleles American Area Binding Biosensor Bone Diseases Cell Lineage ChIP-seq Chromatin Complex DNA Binding DNA-Binding Proteins Data Disease Disease model Dose Epigenetic Process Equilibrium Feedback Fracture Fracture Healing Genes Genetic Gold Hip Fractures Human Kinetics Knock-in Knowledge Mesenchymal Stem Cells Metabolic Bone Diseases Methods Mitogen-Activated Protein Kinases Modeling Molecular Morbidity - disease rate Mus Mutation Names Osteoblasts Osteoclasts Osteogenesis Osteoporosis Ovariectomy Pathology Pathway interactions Phenocopy Postmenopausal Osteoporosis Regulation Reporting Risk Role Series Shapes Signal Transduction Study models Testing WNT Signaling Pathway Woman Work arm base bone bone loss bone mass career development embryonic stem cell experience experimental study in vivo inhibitor/antagonist innovation lifetime risk malignant breast neoplasm medical schools men mortality mouse model novel novel strategies osteoblast differentiation osteoporosis with pathological fracture phosphoproteomics promoter public health relevance research and development response scaffold skeletal skeletal disorder therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Previously we found that the adapter protein SHN3 had a major effect to suppress bone formation by osteoblasts in vivo by inhibiting the activity of the mitogen activated protein kinase (MAPK) ERK. We also identified that WNTs have a previously unidentified activity to activate this SHN3/ERK pathway. Here we seek to make critical extensions of these observations by determining how SHN3 inhibits ERK and how this pathway influences models of skeletal disease in vivo. This area of study is significant as osteoporosis is a highly prevalent disorder of low bone mass and skeletal fragility. Half of all women will experience of an osteoporotic fracture during their lifetime, with each of these fractures incurring significant morbidity and mortality. Based on the profound ability of SHN3 to suppress bone formation, building a better understanding of this pathway will yield new methods to control the activity of osteoblasts to form bone, potentially offering approaches to treat osteoporosis and other disorders of low bone mass. The three overlapping areas to be investigated in this project are: 1. Determine the contribution of SHN3 to skeletal disease models while we have established that SHN3 strongly suppresses bone formation in mice at baseline, the specific contribution of SHN3 to murine models of post-menopausal osteoporosis and skeletal fracture are unknown and will be studied in SHN3-deficient mice. 2. Determine how SHN3 inhibits the ERK pathway in osteoblasts While we have established that SHN3 acts in osteoblasts primarily by suppressing the activity of ERK, the relevant ERK substrates and how SHN3 shapes the kinetics of the ERK response are unknown and will be studied using phosphoproteomics and a quantitative ERK activity biosensor. Determining the relevant substrates of ERK may identify downstream effectors of the SHN3/ERK pathway that are themselves amenable to therapeutic targeting. Additionally, the mechanism for the activation of the ERK/SHN3 pathway by WNTs is unknown, and this will be addressed by a focused screen to determine the MAP3K required for proximal activation of this pathway. 3. Determine if chromatin acts as a scaffold to orchestrate the activity of the SHN3/ERK complex in osteoblasts. Both SHN3 and ERK have previously been identified to bind DNA, raising the possibility that regulation of ERK activity by SHN3 occurs in chromatin bound complexes. In this aim, a complimentary series of chromatin immunoprecipitation-sequencing experiments will be utilized to explore if chromatin orchestrates the function of the SHN3/ERK pathway.

描述（由适用提供）：以前我们发现，衔接蛋白SHN3通过抑制有丝分裂原活化蛋白激酶（MAPK）ERK的活性来抑制成骨细胞的骨骼形成具有重要作用。我们还确定WNT具有先前未鉴定的活动来激活此SHN3/ERK途径。在这里，我们试图通过确定SHN3如何抑制ERK以及该途径如何影响体内骨骼病模型来对这些观察结果进行关键扩展。该研究领域的意义很大，因为骨质疏松症是一种高度普遍的骨质和骨骼脆弱性的疾病。一半的女性将在一生中经历骨质疏松剂，每种骨折都会产生明显的发病率和死亡率。基于SHN3抑制骨形成的深刻能力，对该途径有更好的理解将产生新的方法来控制成骨细胞形成骨骼的活性，并可能提供治疗骨质疏松症和其他低骨质疾病的方法。在该项目中要研究的三个重叠区域是：1。确定SHN3对骨骼疾病模型的贡献，而我们已经确定SHN3在基线下强烈抑制小鼠的骨形成，SHN3对鼠类绝经后的鼠模型的特定贡献和shn3的鼠标均无关。已经确定，SHN3通过抑制ERK的活性，相关的ERK底物以及SHN3如何形成ERK响应的动力学，并将使用磷蛋白组学和定量ERK活性生物传感器研究。确定ERK的相关底物可以识别SHN3/ERK途径的下游效应，而SHN3/ERK途径本身也是治疗靶向的。此外，Wnts激活ERK/SHN3途径的机制尚不清楚，这将通过集中屏幕来解决此途径近端激活所需的MAP3K。 3。确定染色质是否充当支架，以在成骨细胞中编排SHN3/ERK复合物的活性。先前已经鉴定出SHN3和ERK结合DNA，从而提高了SHN3对ERK活性进行调节的可能性。在此目标中，将利用一系列免费的染色质免疫沉淀实验实验来探索染色质是否策划了SHN3/ERK途径的功能。