Formation and function of lamellipodial morphology in 3D microenvironments

项目摘要 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，将他们的发现传达给T细胞和其他适应性免疫细胞 系统。这些专业的迁徙和搜索者是人类免疫力的关键组成部分，他们的 迁徙是多种病原体的目标或劫持，包括一些痘和疱疹病毒，结核病， 还有炭疽热。由于树突状细胞可以激活细胞毒性T细胞攻击癌细胞，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，通过延伸迁移 片状脂溢症。片状脂膜是一种薄的平面突起，已被广泛研究用于细胞迁移。 在2D曲面上，例如玻璃盖片。树突状细胞使用板脂找到一条穿过拥挤的3D的路径 环境和进入淋巴管。片状脂体和组成它们的肌动蛋白网络有 已经研究了几十年了。然而，大多数分子上详细的板脂调节和功能模型 是从研究2D表面上的细胞衍生出来的，所以我们仍然不知道细胞是如何启动和延伸的 3D环境中的片状脂蛋白。Driscoll博士将研究肌动蛋白核素是如何组织起来产生薄片的- 像片状脂肪在没有表面的情况下引导它们的生成，以及肌动蛋白是如何 核子组织起来，在拥挤的3D环境中引导片状脂体的延伸。作为…的典范 树突状细胞在周围组织中的迁移，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，如共聚焦显微镜，不能在3D胶原蛋白中成像细胞 具有测量肌动蛋白核仁的组织所需的空间和时间分辨率 片状脂溢症。然而，最近发展起来的技术，如光片显微镜，才刚刚开始 能够做到这一点。由于光片显微镜可以容易地产生尺寸超过1TB的3D电影， 解释甚至简单地可视化如此大量的数据需要复杂的计算 工作流程。尽管Driscoll博士最近开发了分析光片显微镜的计算工具 为了利用这些工具，她需要在制造和使用光片显微镜方面进行进一步的培训。这个 德州大学西南分校的Danuser和Fiolka实验室是获得这种培训的理想地点。高登兹·丹尼瑟博士 是开发计算机视觉工具来解决细胞生物学问题的专家，而雷托·菲奥卡博士是 专门从事3D系统的薄片显微镜研究。Driscoll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。 总而言之，Driscoll博士将把光片显微镜、3D图像分析和分子生物学结合起来 确定片状脂体如何在3D环境中形成和发挥作用的技术。

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.