MECHANOTRANSDUCTION IN BONE VIA OSCILLATING FLUID FLOW

通过振荡流体流进行骨骼中的机械传导

• 批准号: 7626276
• 负责人: Christopher Rae Jacobs
• 金额: $ 32.3万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-15 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7626276
• 关键词: Actins; Activities of Daily Living; Adhesions; Affect; Bad protein; Behavior; Binding; Biochemical; Bone Diseases; Bone Tissue; Calcium; Calcium Signaling; Cell physiology; Cell surface; Cell-Matrix Junction; Cells; Classification; Custom; Cytoskeleton; Dinoprostone; Ensure; Evaluation; Exposure to; Focal Adhesion Kinase 1; Focal Adhesions; Frequencies; Funding; Genes; Goals; Image Analysis; In Vitro; Integrins; Intervention; Investigation; Knowledge; Laboratories; Lead; Liquid substance; Lung; MAP Kinase Gene; MAPK14 gene; Mechanics; Mediating; Membrane; Metabolism; Microfilaments; Microfluidics; Mitogen-Activated Protein Kinases; Modeling; Molecular; Monitor; Osteoblasts; Osteocytes; Outcome; Pathway interactions; Phosphorylation; Physical activity; Physiological; Point Mutation; Process; Proteins; Proteoglycan; Regulation; Role; SRC gene; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Solutions; Structural Protein; Structure; Supporting Cell; System; Testing; Tissue Engineering; Tyrosine; Tyrosine Phosphorylation; base; bone; bone cell; cellular transduction; design; experience; fluid flow; genetic manipulation; inhibitor/antagonist; monolayer; neuronal cell body; novel; osteogenic; protein protein interaction; release of sequestered calcium ion into cytoplasm; response; shear stress; tool; transmission process

DESCRIPTION (provided by applicant): In the prior funding period we obtained results strongly suggesting that oscillatory fluid flow due to loading is an important cellular physical signal for both osteoblasts and osteocytes. Utilizing our custom built dynamic flow system, we were able to show that oscillatory fluid flow can regulate cell metabolism via intracellular calcium mobilization, prostaglandin E2 release, and MAP kinase activity in the absence of other physical or biochemical signals. However, we have not uncovered the molecular mechanotransduction mechanism activated by oscillatory fluid flow. Candidates can be expected to experience load due to flow and have biochemical signaling potential. This conceptual model is supported by our observation made in the prior funding period that degradation of membrane proteoglycans extending into the flow field has a dramatic effect on the response to flow. Also, we have preliminary indications that actin and focal adhesion kinase (FAK) are involved fluid flow induced signaling. Thus, the central hypothesis of this five year project is that oscillatory fluid flow regulates bone cell metabolism via a molecular mechanism involving forces experienced by the cytoskeleton and transmitted through focal adhesion sites to integrins. To test this hypothesis we will undertake a systematic multilevel evaluation of cell structural proteins to include actin, integrins, and linker proteins both in terms of the effect of oscillatory flow on these proteins (aim 1) and the role of each in transducing the response to flow (aim 2). Additionally, strong evidence from our laboratory and others suggests specific involvement of focal adhesion kinase (FAK) tyrosine phosphorylation. This combined with recently developed molecular tools targeting FAK phosphorylation motivate us to perform a more in-depth investigation of two specific FAK signal pathways (aim 3). Finally, utilizing a novel microfabricated flow chamber, we will determine if the osteocyte process is a specialized structure with enhanced sensitivity to fluid shear forces (aim 4).

描述（由申请人提供）：在之前的资助期间，我们获得的结果强烈表明，由于负载而产生的振荡流体流动对于成骨细胞和骨细胞来说都是重要的细胞物理信号。利用我们定制的动态流动系统，我们能够证明，在没有其他物理或生化信号的情况下，振荡流体流动可以通过细胞内钙动员、前列腺素 E2 释放和 MAP 激酶活性来调节细胞代谢。然而，我们还没有发现振荡流体流动激活的分子力传导机制。候选人预计会因流动而承受负荷，并具有生化信号潜力。这一概念模型得到了我们在之前资助期间的观察的支持，即延伸到流场的膜蛋白聚糖的降解对流动响应具有显着影响。此外，我们有初步迹象表明肌动蛋白和粘着斑激酶 (FAK) 涉及流体流动诱导的信号传导。因此，这个为期五年的项目的中心假设是，振荡流体流通过分子机制调节骨细胞代谢，该分子机制涉及细胞骨架所经历的力并通过粘着斑位点传递给整合素。为了检验这一假设，我们将对细胞结构蛋白（包括肌动蛋白、整合素和连接蛋白）进行系统性多层次评估，评估振荡流对这些蛋白质的影响（目标 1）以及每种蛋白质在转导流动响应中的作用（目标 2）。此外，来自我们实验室和其他实验室的有力证据表明，粘着斑激酶 (FAK) 酪氨酸磷酸化有特定参与。这与最近开发的针对 FAK 磷酸化的分子工具相结合，促使我们对两个特定的 FAK 信号通路进行更深入的研究（目标 3）。最后，利用新型微加工流动室，我们将确定骨细胞过程是否是一种对流体剪切力具有增强敏感性的特殊结构（目标 4）。

项目成果

A role for the primary cilium in paracrine signaling between mechanically stimulated osteocytes and mesenchymal stem cells.

• DOI: 10.1016/j.bbrc.2011.07.072
• 发表时间: 2011-08-19
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Hoey, David A.;Kelly, Daniel J.;Jacobs, Christopher R.
• 通讯作者: Jacobs, Christopher R.

Visualizing in vivo liposomal drug delivery in real-time.

实时可视化体内脂质体药物输送。

• DOI: 10.1080/10611860701538651
• 发表时间: 2007
• 期刊: Journal of drug targeting
• 影响因子: 4.5
• 作者: Kim,Jae-Beom;Leucht,Philipp;Morrell,NathanT;Schwettman,HAlan;Helms,JillA
• 通讯作者: Helms,JillA

A microstructurally informed model for the mechanical response of three-dimensional actin networks.

• DOI: 10.1080/10255840801888686
• 发表时间: 2008-08
• 期刊: Computer methods in biomechanics and biomedical engineering
• 影响因子: 1.6
• 作者: Kwon RY;Lew AJ;Jacobs CR
• 通讯作者: Jacobs CR

Non-canonical Wnt signaling and N-cadherin related beta-catenin signaling play a role in mechanically induced osteogenic cell fate.

• DOI: 10.1371/journal.pone.0005388
• 发表时间: 2009
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Arnsdorf EJ;Tummala P;Jacobs CR
• 通讯作者: Jacobs CR

The epigenetic mechanism of mechanically induced osteogenic differentiation.

• DOI: 10.1016/j.jbiomech.2010.07.033
• 发表时间: 2010-11-16
• 期刊: JOURNAL OF BIOMECHANICS
• 影响因子: 2.4
• 作者: Arnsdorf, Emily J.;Tummala, Padmaja;Castillo, Alesha B.;Zhang, Fan;Jacobs, Christopher R.
• 通讯作者: Jacobs, Christopher R.