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Nanotopographic modulation of B cell signaling activation

B 细胞信号传导激活的纳米拓扑调节

基本信息

批准号：
9281650
负责人：
Arpita Upadhyaya
金额：
$ 18.74万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9281650
关键词：
Actins Actomyosin Antibody Response Antigen Presentation Antigen-Presenting Cells Antigens B-Cell Activation B-Cell Receptor Binding B-Lymphocytes Behavior Biological Biomedical Engineering Cells Cellular Morphology Communicable Diseases Complex Cues Cytoskeleton Dendritic Cells Development Devices Dissection Engineering Environment Goals Immune Immune System Diseases Immune response Immunity Immunotherapy Knowledge Ligands Light Liquid substance Lymphocyte Malignant Neoplasms Mechanics Mediating Membrane Methods Morphology Movement Myosin ATPase Nanostructures Nanotopography Nature Optics Pattern Property Proteins Receptor Activation Receptor Signaling Receptors, Antigen, B-Cell Regulation Regulatory Pathway Resolution Role Signal Transduction Stem cells Surface Technology Testing Therapeutic Vaccines Work base cell behavior cellular imaging design fluorescence imaging genetic regulatory protein high resolution imaging in vivo innovation insight knockout gene lithography live cell imaging microbial nano nanoparticle nanopattern nanosensors nanostructured novel pathogen physical property prevent receptor response small molecule inhibitor vaccine development

项目摘要

B cell-mediated antibody responses are an essential component of immunity and the main target of vaccine development. B cell signaling activation is triggered by the binding of B cell receptors (BCR) with antigen displayed on the surface of professional antigen presenting cells. Although soluble antigen are able to activate B cells, recent studies have shown that surface anchored antigens are significantly more efficient in triggering B cell activation. Consequently, the physical nature of antigen presentation and the mechanical environment of B cells are likely important for BCR activation. In particular, B cells encounter antigen on APC, which possess complex surfaces with convoluted topographies, a fluid membrane and deformable cell bodies. Historically, planar substrates have been used to study immune cell signaling and investigate the mechanisms behind signal initiation. Previous work has shown that many behaviors of adherent and stem cells, such as morphology, movement and differentiation, are modulated by surface topography. These studies demonstrate that cells are able to respond to topographical cues, which in turn influences cell signaling. However, the question of topographical modulation of signaling in immune cells has not been previously studied. Our preliminary studies suggest that B cell signaling and actin dynamics are both influenced by the nanotopography of the antigen-presenting surface. The central hypothesis of this project is that substrate topography influences B signaling and activation. Furthermore, cytoskeletal dynamics at the interface enable the cell to sense topography in order to generate an appropriate signaling response. We will test this hypothesis by using novel nanotopographic surfaces that allow high resolution fluorescence imaging to examine actin cytoskeletal dynamics, BCR signaling and actin regulators in B cells. Aim 1 will examine the effects of substrate nanotopography on B-cell morphology, actin reorganization and signaling activation. We will use nonlinear optical lithography to fabricate novel surfaces with nanotopographic patterns on materials that permit high-resolution live cell imaging of the cell-substrate interface. Using these substrates, we will establish that substrate topography is an important modulator of B cell signaling and cytoskeletal dynamics. Aim 2 will define actin remodeling and their upstream regulators that are critical for topography sensing and the control of BCR signaling. To do so, we will perturb the actomyosin cytoskeleton using small-molecule inhibitors and gene knockout of regulatory proteins including WASP and N-WASP in modulating topographic sensing in B cells via actin remodeling. We will further test the role of curvature sensing proteins in topographic sensing by the control of membrane curvature. These studies will provide important new insights into how actin regulatory pathways mediate topographic sensing, giving us another means of tuning B cell signaling with nanotopography. More broadly, this work will shed light on mechanisms by which cells sense their environment by surface receptors, which has considerable implications for biomedicine.

B细胞介导的抗体应答是免疫的重要组成部分，也是疫苗的主要靶点发展B细胞信号传导激活由B细胞受体（BCR）与抗原的结合触发展示在专职抗原呈递细胞的表面。虽然可溶性抗原能够激活 B细胞，最近的研究表明，表面锚定抗原在触发 B细胞活化。因此，抗原递呈的物理性质和抗原递呈的机械环境是不确定的。 B细胞可能对BCR活化很重要。特别地，B细胞遇到APC上的抗原，APC具有复杂的表面，具有复杂的形貌、流体膜和可变形的细胞体。从历史上看，平面基板已被用于研究免疫细胞信号传导和调查背后的机制信号启动以前的研究表明，粘附细胞和干细胞的许多行为，如形态、运动和分化受表面形貌的调节。这些研究证明细胞能够对地形线索做出反应，这反过来又会影响细胞信号。但免疫细胞中信号传导的局部调节问题以前没有研究过。我们初步研究表明，B细胞信号传导和肌动蛋白动力学都受到抗原呈递表面的纳米形貌。这个项目的核心假设是，地形影响B信号和激活。此外，界面处的细胞骨架动力学使细胞能够感测形貌，以便产生适当的信号响应。我们将测试该假设通过使用允许高分辨率荧光成像的新型纳米形貌表面，研究肌动蛋白细胞骨架动力学，BCR信号和B细胞中的肌动蛋白调节因子。目标1将审查基底纳米形貌对B细胞形态、肌动蛋白重组和信号传导激活的影响。我们将使用非线性光学光刻技术在材料上制造具有纳米形貌图案的新型表面其允许细胞-基质界面的高分辨率活细胞成像。使用这些基质，我们将建立基质形貌是B细胞信号传导和细胞骨架动力学重要调节剂。目的2将定义肌动蛋白重塑及其上游调节因子，这些因子对地形感知至关重要，控制BCR信号。为此，我们将使用小分子抑制剂扰乱肌动球蛋白细胞骨架以及包括WASP和N-WASP在内的调节蛋白在调节地形感知中的基因敲除。 B细胞通过肌动蛋白重塑。我们将进一步测试曲率感应蛋白在地形感应中的作用通过控制膜的曲率。这些研究将提供重要的新见解，调节途径介导地形传感，给我们提供了另一种调节B细胞信号传导的方法，纳米形貌学更广泛地说，这项工作将阐明细胞感知环境的机制这对生物医学有很大的影响。