Structural, Molecular and Functional Specialization in Osteocyte Mechanosensing

骨细胞机械传感的结构、分子和功能专业化

基本信息

批准号：
10394277
负责人：
MITCHELL B SCHAFFLER
金额：
$ 62.82万
依托单位：
CITY COLLEGE OF NEW YORK
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-11 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10394277
关键词：
Acute Affect Affinity Androgens Binding Biochemical Bone Diseases Bone Resorption Bone remodeling Calcium Cell Signaling Process Cell physiology Cells Co-Immunoprecipitations Complex Data Effector Cell Elements Endocrine Estrogens Fingers Fluorescence Resonance Energy Transfer Focal Adhesions Frequencies Funding Genes Genetic Models Goals Gonadal Steroid Hormones Hormonal Hormonal Change Hormones Image In Vitro Individual Integrin alphaVbeta3 Integrins Intervention Ion Channel Life Mechanical Stimulation Mechanics Membrane Methods Microscopy Modeling Molecular Mus Mutation Osteoblasts Osteoclasts Osteocytes Osteogenesis Osteoporosis prevention P2X-receptor Pathway interactions Pattern Pharmacology Physiological Process Property Purinoceptor Reporter Reporting Resolution Role Series Signal Pathway Signal Transduction Site Skeleton Structure Surface Plasmon Resonance System T-Type Calcium Channels Techniques Testing United States National Institutes of Health base beta catenin bone bone cell bone fragility bone health experimental study follow-up imaging modality imaging system improved in vivo individual response innovation mechanical load mechanical signal mechanical stimulus mechanotransduction molecular assembly/self assembly mouse genetics multidisciplinary new therapeutic target novel receptor recruit response restoration sensor

项目摘要

ABSTRACT Bone adapts its structure to mechanical loading. This adaption is essential for growing the right skeleton and maintaining its integrity throughout life. Osteocytes are the cells responsible for sensing and coordinating response to mechanical load. Key recent discoveries reported by our group during the last several years established that osteocyte cell processes function as unique mechanosensory elements. Processes are >10-fold more sensitive to mechanical stimuli than osteocyte cell bodies. Moreover, this triggering of Ca2+ signaling from cell processes occurs through a unique complex of aVb3 integrins, membrane channels and receptors, that occur at attachment points to the canalicular walls, and which we call the “Osteocyte mechanosome.” This proposal is based on the global hypothesis that a novel structure localized on osteocyte processes, the osteocyte mechanosome, detects and transduces mechanical signals. To date, we have identified four key osteocyte mechanosome components: αVβ3 integrin, pannexin1, P2X7 receptor (P2X7R) and the CaV3.2 T-type calcium channel. Our multidisciplinary team will test this hypothesis by multiple approaches in each of three aims. In Aim 1 we will combine biochemical techniques (co-immunoprecipitation, surface plasmon resonance) and imaging modalities (FRAP, FRET and STORM super-resolution microscopy) to define comprehensively the structural and dynamic properties of this heretofore unknown transduction complex, the osteocyte mechanosome in osteocytic cells in vitro. In Aim 2 we test how pharmacological and genetic alteration of individual mechanosome components alters upstream (Ca2+) and downstream (to bone) signaling in osteocytic cells in vitro. In Aim 3, we will combine our novel OtGP3 osteocyte Ca2+ reporter mice-in vivo loading/imaging system with pharmacological manipulations to confirm effects of key mechanosome components (as identified in Aims 1 and 2) on osteocyte Ca2+ response and on downstream signaling. We will also use this approach to answer the fundamental question of whether osteocyte Ca2+ responses to mechanical loading altered by loss of constitutive sex hormones (estrogen/androgen) or by anabolic PTH.

抽象的骨骼将其结构适应机械载荷。这种适应对于发展权利至关重要骨骼并保持其一生的完整性。骨细胞是负责的细胞传感和协调对机械负载的响应。我们报告的关键发现是我们的在过去的几年中，小组确定骨细胞细胞过程起着独特的作用力学元素。与机械刺激更敏感的过程比骨细胞细胞体。此外，发生了来自单元过程的Ca2+信号传导的触发通过独特的AVB3整合素，膜通道和接收器的复合物发生在附着点指向管壁，我们称之为“骨细胞机械体”。该提议基于一个全球假设，即新型结构位于骨细胞上过程，骨细胞机械体，检测和传输机械信号。迄今为止，我们已经确定了四个关键的骨细胞机械体成分：αVβ3整合素，pannexin1， P2X7受体（P2X7R）和CAV3.2 T型钙通道。我们的多学科团队将通过三个目标中的每种方法都通过多种方法检验该假设。在AIM 1中，我们将结合生化技术（共免疫沉淀，表面等离子体共振）和成像方式（FRAP，FRET和Storm超分辨率显微镜）以全面定义该迄今未知翻译复合物的结构和动态特性，体外骨细胞中的骨细胞机械体。在AIM 2中，我们测试了药物和单个机械体成分的遗传改变会改变上游（Ca2+）和体外骨细胞中的下游（骨）信号传导。在AIM 3中，我们将结合小说 otgp3骨细胞CA2+报道鼠标的体内载荷/成像系统与药物操纵以确认关键机械体组件的影响（如AIM 1和 2）在骨细胞Ca2+响应和下游信号传导上。我们还将使用这种方法回答骨细胞CA2+对机械加载的反应的基本问题因构成性激素（雌激素/雄激素）或合成代谢PTH的丧失而改变。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Estrogen depletion on In vivo osteocyte calcium signaling responses to mechanical loading.

DOI：
10.1016/j.bone.2021.116072
发表时间：
2021-11
期刊：
Bone
影响因子：
4.1
作者：
Lewis KJ;Cabahug-Zuckerman P;Boorman-Padgett JF;Basta-Pljakic J;Louie J;Stephen S;Spray DC;Thi MM;Seref-Ferlengez Z;Majeska RJ;Weinbaum S;Schaffler MB
通讯作者：
Schaffler MB

A Bisphosphonate With a Low Hydroxyapatite Binding Affinity Prevents Bone Loss in Mice After Ovariectomy and Reverses Rapidly With Treatment Cessation.

DOI：
10.1002/jbm4.10476
发表时间：
2021-04
期刊：
JBMR plus
影响因子：
3.8
作者：
Coffman AA;Basta-Pljakic J;Guerra RM;Ebetino FH;Lundy MW;Majeska RJ;Schaffler MB
通讯作者：
Schaffler MB

A multiscale fluidic device for the study of dendrite-mediated cell to cell communication.

DOI：
10.1007/s10544-017-0212-1
发表时间：
2017-08-08
期刊：
Biomedical microdevices
影响因子：
2.8
作者：
McCutcheon S;Majeska R;Schaffler M;Vazquez M
通讯作者：
Vazquez M

Induction of somatopause in adult mice compromises bone morphology and exacerbates bone loss during aging.

DOI：
10.1111/acel.13505
发表时间：
2021-12
期刊：
Aging cell
影响因子：
7.8
作者：
Dixit M;Duran-Ortiz S;Yildirim G;Poudel SB;Louis LD;Bartke A;Schaffler MB;Kopchick JJ;Yakar S
通讯作者：
Yakar S

Role of pannexin 1 channels in load-induced skeletal response.

pannexin 1 通道在负荷诱导的骨骼反应中的作用。

DOI：
10.1111/nyas.13914
发表时间：
2019
期刊：
Annals of the New York Academy of Sciences
影响因子：
5.2
作者：
Seref-Ferlengez,Zeynep;Urban-Maldonado,Marcia;Sun,HuiB;Schaffler,MitchellB;Suadicani,SylviaO;Thi,MiaM
通讯作者：
Thi,MiaM