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：
10792225
负责人：
Meghan Katrien Driscoll
金额：
$ 4.78万
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

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 lym- phatic vessels. Lamellipodia and the actin network that composes them have been studied for decades. How- ever, 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. In this project, we 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 exten- sion of lamellipodia within crowded 3D environments. As a model of dendritic cell migration through peripheral tissues, we will study 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 produce massive datasets, interpreting and even simply visualizing such large amounts of data requires sophisticated computing workflows. We will develop the needed computational workflows as we investigate 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到淋巴结，将它们的发现传达给 T 细胞和适应性免疫系统的其他细胞。这些专业的迁徙者和搜寻者是人类免疫力的重要组成部分，他们的迁徙是被多种病原体攻击或劫持，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中发挥着作用免疫治疗策略。许多细胞，包括树突细胞，通过延伸片状伪足来迁移。片状伪足是薄的平面突起，已被广泛研究细胞在 2D 表面（例如玻璃）上迁移盖玻片。树突状细胞利用板状伪足寻找穿过拥挤的 3D 环境的路径并进入淋巴管。食管。片状伪足和组成它们的肌动蛋白网络已经被研究了几十年。如何- 迄今为止，大多数板状伪足调节和功能的分子详细模型都源自研究细胞在 2D 表面上，所以我们仍然不知道细胞如何在 3D 环境中启动和扩展片状伪足。在这个项目中，我们将研究肌动蛋白成核剂如何组织以产生片状片状足形态缺乏表面来指导它们的生成，以及肌动蛋白成核剂如何组织以指导延伸拥挤的 3D 环境中片状伪足的融合。作为树突状细胞通过外周迁移的模型组织，我们将研究通过 3D 纤维胶原基质的迁移。广泛使用的显微技术，例如共焦显微镜，无法以所需的空间和时间分辨率对 3D 胶原蛋白中的细胞进行成像测量片状伪足中肌动蛋白成核剂的组织。然而，最近开发的技术，例如作为光片显微镜，才刚刚开始能够做到这一点。由于光片显微镜可以产生海量数据集，解释甚至简单地可视化如此大量的数据都需要复杂的技术计算工作流程。当我们研究片状伪足形式时，我们将开发所需的计算工作流程并在 3D 环境中运行。