Formation and function of lamellipodial morphology in 3D microenvironments

3D 微环境中片状足形态的形成和功能

基本信息

批准号：
10733474
负责人：
Meghan Katrien Driscoll
金额：
$ 24.9万
依托单位：
UNIVERSITY OF MINNESOTA
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：
2018-04-01 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10733474
关键词：
3-Dimensional Actins Adaptive Immune System Address Adhesions Affect Anthrax disease Antigens Biochemical Biological Assay Biological Models Cell physiology Cells Cellular Morphology Cellular biology Collagen Collagen Fiber Color Communication Complex Computer Vision Systems Confocal Microscopy Coupling Crowding Cytotoxic T-Lymphocytes Data Dendritic Cells Diffusion Elements Embryonic Development Environment Exhibits Fibroblasts Future Generations Geometry Glass Grant Human Image Image Analysis Immune Immune system Immunity Investigation Laboratories Lead Light Location Macrophage Maintenance Measures Membrane Microscope Microscopic Microscopy Modeling Molecular Molecular Biology Techniques Morphology Neoplasm Metastasis Neural Crest Cell Pathologic Processes Penetration Peripheral Phase Physiological Processes Polymers Property Proteins Regulation Resolution Sentinel Shapes Signal Transduction Slice Structure Surface System T-Lymphocyte Techniques Testing Thinness Three-Dimensional Image Tissues Training Tuberculosis Virus Visualization Work cancer cell cancer immunotherapy cell motility cell type cellular imaging computerized tools imaging modality immune function in vivo light microscopy lymph nodes lymphatic vessel microscopic imaging migration movie novel pathogen polymerization predictive modeling recruit spatiotemporal temporal measurement tool wound healing

项目摘要

Project Summary Dendritic cells are the sentinels of the immune system. They patrol the body looking for antigens and then migrate to a lymph node to communicate what they found to T cells and other cells of the adaptive immune system. These professional migrators and searchers are a critical component of human immunity, and their migration is targeted or hijacked by multiple pathogens including some pox and herpes viruses, tuberculosis, and anthrax. Since dendritic cells can activate cytotoxic T cells to attack cancer cells, their migration also plays a role in cancer immunotherapy strategies. Many cells, including dendritic cells, migrate by extending lamellipodia. Lamellipodia are thin, planar protrusions that have been extensively studied for cells migrating on 2D surfaces, such as glass coverslips. Dendritic cells use lamellipodia to find a path through crowded 3D environments and to enter lymphatic vessels. Lamellipodia and the actin network that composes them have been studied for decades. However, most molecularly detailed models of lamellipodia regulation and function were derived from studying cells on 2D surfaces, so we still do not know how cells initiate and extend lamellipodia in 3D environments. Dr. Driscoll will investigate how actin nucleators organize to generate sheet- like lamellipodial morphologies in the absence of a surface to guide their generation, as well as how actin nucleators organize to direct the extension of lamellipodia within crowded 3D environments. As a model of dendritic cell migration through peripheral tissues, she will study their migration though 3D fibrous collagen matrices. Widely available microscopic techniques, such as confocal microscopy, cannot image cells in 3D collagen with the spatial and temporal resolution required to measure the organization of actin nucleators in lamellipodia. However, recently developed techniques, such as light-sheet microscopy, are just beginning to be able to do so. Since light-sheet microscopes can easily produce 3D movies exceeding 1TB in size, interpreting and even simply visualizing such large amounts of data requires sophisticated computing workflows. Although Dr. Driscoll has recently developed computational tools to analyze light-sheet microscopy images, to utilize these tools she needs further training in building and using light-sheet microscopes. The Danuser and Fiolka labs at UT Southwestern are ideal locations to obtain this training. Dr. Gaudenz Danuser is an expert at developing computer vision tools to address cell biology questions, whereas Dr. Reto Fiolka is specialized in light-sheet microscopy of 3D systems. The training Dr. Driscoll receives will enable her to lead an independent laboratory that focuses on how cells migrate through and interact with their 3D environment. In summary, Dr. Driscoll will integrate light-sheet microscopy, 3D image analysis, and molecular biology techniques to determine how lamellipodia form and function in 3D environments.

项目概要树突状细胞是免疫系统的哨兵。它们在体内巡逻寻找抗原，然后迁移至淋巴结，将其发现的信息传达给 T 细胞和其他适应性免疫细胞系统。这些专业的迁徙者和搜索者是人类免疫力的重要组成部分，他们的迁徙是多种病原体的目标或劫持，包括一些痘病毒和疱疹病毒、结核病、和炭疽病。由于树突状细胞可以激活细胞毒性 T 细胞来攻击癌细胞，因此它们的迁移也在癌症免疫治疗策略中发挥作用。许多细胞，包括树突状细胞，通过延伸来迁移片状伪足。片状伪足是薄的平面突起，已被广泛研究细胞迁移在 2D 表面上，例如玻璃盖玻片。树突状细胞利用片状伪足在拥挤的 3D 空间中寻找路径环境并进入淋巴管。板状伪足和组成它们的肌动蛋白网络具有研究了几十年。然而，大多数板状伪足调节和功能的分子详细模型来自研究二维表面上的细胞，所以我们仍然不知道细胞如何启动和延伸 3D 环境中的片状伪足。德里斯科尔博士将研究肌动蛋白成核剂如何组织以产生片状- 像在没有表面指导其生成的情况下的片状足形态，以及肌动蛋白如何成核剂组织起来，在拥挤的 3D 环境中引导片状伪足的延伸。作为一个模型树突状细胞通过外周组织迁移，她将研究它们通过 3D 纤维胶原蛋白的迁移矩阵。广泛使用的显微技术（例如共焦显微镜）无法对 3D 胶原蛋白中的细胞进行成像具有测量肌动蛋白成核剂组织所需的空间和时间分辨率片状伪足。然而，最近开发的技术，例如光片显微镜，才刚刚开始能够这样做。由于光片显微镜可以轻松制作超过 1TB 大小的 3D 电影，解释甚至简单地可视化如此大量的数据都需要复杂的计算工作流程。尽管德里斯科尔博士最近开发了计算工具来分析光片显微镜图像，为了使用这些工具，她需要进一步接受构建和使用光片显微镜的培训。这 UT 西南大学的 Danuser 和 Fiolka 实验室是获得此培训的理想地点。高登兹·丹努瑟博士是开发计算机视觉工具来解决细胞生物学问题的专家，而 Reto Fiolka 博士是专门研究 3D 系统的光片显微镜。德里斯科尔博士接受的培训将使她能够领导一个独立的实验室，专注于细胞如何在 3D 环境中迁移并与其相互作用。总之，Driscoll 博士将整合光片显微镜、3D 图像分析和分子生物学确定片状伪足如何在 3D 环境中形成和发挥作用的技术。