Mechanisms of regulation of lymphocyte migration by actin cytoskeletal effectors

肌动蛋白细胞骨架效应器调节淋巴细胞迁移的机制

基本信息

批准号：
10583309
负责人：
Jordan Jacobelli
金额：
$ 49.2万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-23 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10583309
关键词：
3-Dimensional Actins Adhesions Adhesives Adopted Autoimmune Autoimmunity Back Biological Models Blood Vessels Cell Communication Cell Nucleus Cells Characteristics Collagen Complement Complex Cues Cytoskeletal Proteins Cytoskeleton DNA Sequence Alteration Data Disease Embryonic Development Endothelial Cells Environment Environmental Risk Factor Extracellular Matrix Extravasation FMNL1 gene Face Family Generations Goals Homologous Gene Imaging Techniques Immune Immune response Immune system In Vitro Inflammatory Knowledge Ligands Location Lymphocyte Malignant Neoplasms Mediating Membrane Methods Microtubules Modeling Neoplasm Metastasis Nonmuscle Myosin Type IIA Pathologic Physiological Physiological Processes Play Protein Family Proteins Regulation Role Shapes T-Lymphocyte Testing Therapeutic Tissues Traction Force Microscopy Vascular Endothelium Work biophysical properties cancer cell cell motility cell type chemokine constriction immune function immune system function in vivo insight interstitial lymph nodes mechanical force migration novel pathogen three-dimensional modeling trafficking two photon microscopy wound healing

3-Dimensional Actins Adhesions Adhesives Adopted Autoimmune Autoimmunity Back Biological Models Blood Vessels Cell Communication Cell Nucleus Cells Characteristics Collagen Complement Complex Cues Cytoskeletal Proteins Cytoskeleton DNA Sequence Alteration Data Disease Embryonic Development Endothelial Cells Environment Environmental Risk Factor Extracellular Matrix Extravasation FMNL1 gene Face Family Generations Goals Homologous Gene Imaging Techniques Immune Immune response Immune system In Vitro Inflammatory Knowledge Ligands Location Lymphocyte Malignant Neoplasms Mediating Membrane Methods Microtubules Modeling Neoplasm Metastasis Nonmuscle Myosin Type IIA Pathologic Physiological Physiological Processes Play Protein Family Proteins Regulation Role Shapes T-Lymphocyte Testing Therapeutic Tissues Traction Force Microscopy Vascular Endothelium Work biophysical properties cancer cell cell motility cell type chemokine constriction immune function immune system function in vivo insight interstitial lymph nodes mechanical force migration novel pathogen three-dimensional modeling trafficking two photon microscopy wound healing

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项目摘要

Lymphocytes must correctly localize to mount effective immune responses. To do this, lymphocytes migrate through tissue environments with very different biophysical characteristics, including extravasation from blood vessels and crawling through cell-packed or extracellular matrix-rich tissues. To navigate through these diverse environments, lymphocytes squeeze through constrictions and migrate in low- or high-adhesive environments. However, there is a key gap in the understanding of how specific cytoskeletal effectors regulate force generation, shape changes, and cell-cell or cell-matrix interactions during lymphocyte migration in different settings. Given their relevance to immune function, primary T lymphocytes are a highly significant model to investigate cytoskeletal regulation of the varying modes of amoeboid cell motility in three-dimensional (3D) environments. Formin family proteins are cytoskeletal effectors involved in mediating actin network remodeling. Formin-like-1 (FMNL1) and Diaphanous-homologue-1 (mDia1) are the two most highly expressed Formins in T cells. We recently determined that FMNL1 is required for T cell transendothelial migration (TEM) and trafficking to inflamed tissues. Interestingly, our preliminary data support that FMNL1 and mDia1 have distinct functions in T cell migration through confined environments. Our goal is to achieve a more comprehensive understanding of the mechanisms by which the cytoskeleton enables migration through diverse tissue environments by determining the mode of action of Formin proteins in T cell motility. We will investigate the mechanisms by which Formins generate mechanical forces during T cell migration, the contribution of Formins in promoting T cell nucleus passage through constrictions, and how Formins regulate T cell motility in vivo. We will also determine if FMNL1 and mDia1 act independently or in concert with the Arp2/3 complex and/or Myosin-IIA. Our hypothesis is that to promote migration through complex environments FMNL1 mediates force generation from the back of the T cell while mDia1 creates force and membrane protrusions at the cell front. To test this hypothesis, we will use a multi-faceted approach, including genetic/mutational approaches and advanced imaging techniques in complementary model systems in vitro and physiological environments in vivo. Aim 1: Determine the mechanisms by which FMNL1 and mDia1 promote T cell transendothelial migration. Aim 2: Determine how FMNL1 and mDia1 regulate T cell motility within 3D environments with diverse biophysical characteristics. Aim 3: Define how Formins regulate T cell extravasation and interstitial motility in vivo. Overall, our studies are significant in that they will advance our knowledge of T cell migration by providing new data to determine the mechanism of action of Formins in T cell motility and if they cooperate with Myosin-IIA to promote migration through environments with varied biophysical characteristics. Thus, this work has the potential to provide important insight into novel ways to therapeutically modulate lymphocyte migration, such as in autoimmune settings and inflammatory diseases.

淋巴细胞必须正确定位以实现有效的免疫反应。为此，淋巴细胞通过具有截然不同的生物物理特征的组织环境迁移，包括渗出从血管和爬行细胞包装或细胞外基质的组织中爬行。导航这些不同的环境，淋巴细胞通过收缩挤压并在低粘合剂或高粘合剂中迁移环境。但是，了解特定细胞骨架效应子如何调节的关键差距淋巴细胞迁移期间的力产生，形状变化以及细胞细胞或细胞 - 基质相互作用不同的设置。鉴于它们与免疫功能相关，一级T淋巴细胞是一种非常重要的模型研究三维中变化模式的各种模式的细胞骨架调节（3D）环境。 formin家族蛋白是介导肌动蛋白网络涉及的细胞骨架效应子重塑。 formin样-1（FMNL1）和diaphanous-Holomogue-1（MDIA1）是两个最高表达的 T细胞中的造型。我们最近确定FMNL1是T细胞跨内皮迁移（TEM）所必需的并贩运发炎的组织。有趣的是，我们的初步数据支持FMNL1和MDIA1具有 T细胞通过限制环境的迁移中的不同功能。我们的目标是实现更多对细胞骨架可以通过多种方式迁移的机制的全面理解通过确定formin蛋白在T细胞运动中的作用方式通过组织环境。我们将调查 formins在T细胞迁移过程中产生机械力的机制，造型通过收缩促进T细胞核通过的传递，以及造型如何调节T细胞运动性体内。我们还将确定FMNL1和MDIA1是独立起作用还是与ARP2/3配合和/或肌球蛋白-IIA。我们的假设是促进通过复杂环境的迁移fmnl1 从T细胞的背面介导力产生，而MDIA1在细胞正面。为了检验这一假设，我们将使用多方面的方法，包括遗传/突变互补模型系统中的方法和高级成像技术在体外和生理上体内环境。目标1：确定FMNL1和MDIA1促进T细胞的机制跨内皮迁移。 AIM 2：确定FMNL1和MDIA1如何调节3D内的T细胞运动具有不同生物物理特征的环境。 AIM 3：定义formins如何调节T细胞渗出和体内的间质运动。总体而言，我们的研究很重要，因为它们将提高我们对T的了解通过提供新数据来确定formins在T细胞运动中的作用机理，以及IS 他们与肌球蛋白-IIA合作，通过不同的生物物理环境促进迁移特征。因此，这项工作有潜力提供对新颖方法进行治疗方法的重要见解调节淋巴细胞迁移，例如在自身免疫性环境和炎症性疾病中。