Microsystems to Manipulate Fibroblast Chemotaxis

操纵成纤维细胞趋化性的微系统

基本信息

批准号：
7025786
负责人：
MARIBEL VAZQUEZ
金额：
$ 14.94万
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：
2005
资助国家：
美国
起止时间：
2005-03-01 至 2008-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7025786
关键词：
affinity labeling binding proteins biological signal transduction biotechnology chemotaxis fibroblasts growth factor receptors immunoconjugates immunofluorescence technique method development mitogen activated protein kinase nanotechnology platelet derived growth factor tissue engineering wound healing

项目摘要

DESCRIPTION (provided by applicant): During tissue repair, fibroblast cells chemotax, i.e. migrate in the direction of extracellular chemical gradients, using the numerous biochemical receptors distributed on their surface. This project proposes a new, innovative approach to chemotaxis study by developing a model system and methodology to identify the causality between extracellular ligands imposed onto a fibroblast cell, and the consequent redistribution of intracellular proteins needed to initiate chemotaxis-driving signal cascades. It is our hypothesis that the Mitogen-Activated Protein Kinase (MAP) cascade regulates fibroblast migration by influencing the distribution of the docking protein, Growth Factor Receptor Bound-2 (GRB2), which binds to the surface receptors activated by the known chemoattractant ligand, Platelet-Derived Growth Factor Beta (PDGFB). The current project will test this hypothesis via experiments that utilize our own transparent, single-cell, chemotaxis system nicknamed the microLane, to impose one-dimensional concentration gradients of PDGFB across bovine ligament fibroblast cells. GRB2 molecules of the fibroblasts tested within our system will be labeled and mapped using the fluorescent signatures of nanocrystal Bioconjugates (Q-Dots) bound to GRB2 antibodies, apriori. Quantitative results describing GRB2 redistribution as a function of imposed PDGFB gradients will develop a new and unprecedented method to manipulate fibroblast chemotaxis during wound healing. The results will impact Human health via clinical development of therapeutic technologies in avascular tissue repair, as well as research development of collagen scaffolds and biomaterials needed to advance wound healing.

描述（由申请人提供）：在组织修复期间，成纤维细胞趋障，即使用分布在其表面上的大量生化受体朝着细胞外化学梯度的方向迁移。该项目通过开发一种模型系统和方法来识别施加到成纤维细胞上的细胞外配体之间的因果关系，并通过开发一种模型系统和方法来提出一种新的创新方法，以及随之而来的细胞内蛋白质的重新分布，以启动趋化性驱动式驱动信号信号信号级联。我们的假设是，有丝分裂原激活的蛋白激酶（MAP）级联反应通过影响对接蛋白，生长因子受体结合2（GRB2）的分布来调节成纤维细胞的迁移，该分布与由已知的化合物配体，血小板衍生的生长因子beta（PDGFB）激活的表面受体。当前的项目将通过使用我们自己的透明，单细胞，趋化性系统的实验来检验这一假设，以昵称为Microlane，以在跨牛蛋白蛋白蛋白蛋白蛋白蛋白蛋白蛋白质量的成纤维细胞中施加PDGFB的一维浓度梯度。在我们系统中测试的成纤维细胞的GRB2分子将使用与GRB2抗体结合的纳米晶体生物缀合物（Q点）的荧光特征进行标记和映射。描述GRB2重新分布作为施加的PDGFB梯度的函数的定量结果将开发出一种新的和前所未有的方法，以在伤口愈合过程中操纵成纤维细胞趋化性。结果将通过血管组织修复中治疗技术的临床开发以及胶原蛋白脚手架的研究开发和促进伤口愈合所需的生物材料的研究。