Bio-Mechanics of Directional Migration of Leukocytes

白细胞定向迁移的生物力学

基本信息

批准号：
9315164
负责人：
RICHARD A FIRTEL
金额：
$ 37.68万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-30 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9315164
关键词：
3-Dimensional Actins Adhesions Anti-Inflammatory Agents Anti-inflammatory Atomic Force Microscopy Autoimmune Diseases Automobile Driving Basement membrane Biochemical Biochemical Process Biomechanics Blood Vessels Cells Characteristics Chronic Collagen Crowding Cytoskeleton Development Dictyostelium Dictyostelium discoideum Dimensions Disease Endothelial Cells Engineering Environment Event Extracellular Matrix Extravasation Fibrinogen Generations Genetic Grant In Vitro Infection Inflammation Inflammatory Inflammatory Bowel Diseases Injury Insulin-Dependent Diabetes Mellitus Integrins Intercellular Junctions Invaded Investigation Lead Leukocytes Light Location Locomotion Measures Mechanics Mediating Microscopy Molecular Multiple Sclerosis Myosin Type II Outcomes Research Pharmacology Phase Play Principal Component Analysis Process Property Recruitment Activity Regimen Research Resolution Rheumatoid Arthritis Role Series Shapes Site Speed Statistical Data Interpretation Stream Stress Structure Surface Techniques Tissues Traction Vascular Endothelial Cell adaptive immune response cell motility design driving force experience experimental study loss of function migration monolayer multidisciplinary novel novel strategies polymerization public health relevance repaired

项目摘要

DESCRIPTION: The innate and adaptive immune response involves the recruitment of leukocytes from the blood stream to the site of infection and inflammation. Upon reaching the location, leukocytes clear invaders and begin the process of digesting and repairing damaged tissues. However, when the body fails to properly regulate the recruitment of leukocytes, the inflammation can become chronic, resulting in irreversible tissue injury and loss of functionality. Rheumatoid arthritis, inflammatory bowel disease, type-1 diabetes, and multiple sclerosis are all examples of autoimmune diseases caused by the uncontrolled recruitment of leukocytes. While much research has been dedicated to the identification of the cascade of specific biochemical processes involved in the recruitment of leukocytes, much less is known about the mechanical events driving their migration, in particular how they generate the necessary traction forces to cross the vascular wall and further traverse the three- dimensional (3-D) extravascular space. Thus, the main objective of this study is to provide the much needed complementary information connecting specific cell molecular processes (i.e., adhesion dynamics, actin turnover, and myosin II contraction) to the generation of cellular forces that regulate leukocyte extravasation and their subsequent directional migration in 3-D extravascular tissues through the use of novel 3D Fourier Traction Force Microscopy (3DFTFM) techniques and genetic and pharmacological manipulations. To achieve this objective, we propose three Specific Aims. We will first characterize the temporal and spatial generation of 3-D traction forces exerted by leukocytes crawling on flat surfaces (Aim 1); we will then investigate the mechanical processes regulating transmigration across the vascular endothelial monolayer and the basement membrane (Aim 2); and finally, we will develop a novel Elastographic 3DTFM to determine both traction stresses and the non-linear material properties of the Extra Cellular Matrix to elucidate the molecular mechanisms regulating the mechanics of leukocytes' chemotactic migration in 3-D environments (Aim 3). The proposed in vitro approach overcomes a number of existing challenges to measuring the 3-D traction forces driving leukocyte extravasation and migration and builds on the extensive experience accumulated by our multidisciplinary team of biologists and engineers who have been studying the mechanics of amoeboid cell migration for the last seven years. The outcome of this research will result in a far more comprehensive understanding of the mechanics of leukocyte motility than that available to date and will have the potential to aid the development of new approaches that could target specific mechanical processes to inhibit (or slow down) leukocyte motility and help in the design of complementary regimens to treat inflammatory diseases.

描述：先天和适应性免疫响应涉及从血流到感染和感染部位募集白细胞。到达位置后，白细胞清除入侵者，开始消化和修复受损坏的组织的过程。但是，当人体无法正确调节白细胞的募集时，炎症可能会变得慢性，从而导致不可逆转的组织损伤和功能丧失。类风湿关节炎，炎症性肠病，1型糖尿病和多发性硬化症都是由白细胞不受控制募集引起的自身免疫性疾病的例子。尽管已经进行了许多研究，以识别募集白细胞涉及的特定生化过程的级联，但对于驱动其迁移的机械事件，特别是它们如何产生必要的牵引力以越过血管壁并进一步传播三维（3-D-D-D）跨膜间空间，少得多。这是这项研究的主要目的是为连接特定细胞分子过程（即粘合动力学，肌动蛋白周转和肌球蛋白II的收缩）提供急需的互补信息，从而产生细胞力的产生，这些细胞力的产生，这些力量通过使用3D旋转力（3D旋转力）（3DDDDD扭转力）（3DDDDD型尿布术）来调节白细胞渗入及其随后的方向迁移的细胞力。药物操纵。为了实现这一目标，我们提出了三个具体目标。我们将首先表征白细胞在平坦表面上爬行的临时和空间生成的3-D牵引力（AIM 1）；然后，我们将研究跨血管内皮单层和基底膜的机械过程（AIM 2）；最后，我们将开发一种新型的弹性3DTFM，以确定额外细胞基质的牵引力和非线性材料特性，以阐明在3-D环境中衡量白细胞趋化性迁移机制的分子机制（AIM 3）。提出的体外方法克服了许多现有的挑战，用于衡量驱动白细胞渗出和迁移的3D牵引力，并建立在我们多学科的生物学家和工程师团队积累的丰富经验，他们在过去的七年中一直研究Amoeboid细胞迁移的机制。这项研究的结果将使人们对白细胞运动的机制有更全面的了解，而不是迄今为止可用的机制，并且有可能帮助开发新方法，这些方法可能针对特定的机械过程，以抑制（或放慢）白细胞运动并帮助治疗炎性疾病的互补方案。