Deciphering the mechanism of SHIP1 regulation in human neutrophils

破译 SHIP1 在人类中性粒细胞中的调节机制

基本信息

批准号：
10280943
负责人：
Scott David Hansen
金额：
$ 29.54万
依托单位：
UNIVERSITY OF OREGON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10280943
关键词：
1-Phosphatidylinositol 3-Kinase Actins Architecture Bacteria Behavior Binding Biochemical Biochemistry Biological Models Biophysics Biosensor C-terminal CRISPR interference Cell Line Cell Polarity Cell Surface Receptors Cell membrane Cell physiology Cells Cellular biology Chemicals Chemotaxis Clustered Regularly Interspaced Short Palindromic Repeats Coin Communication Cues Decision Making Engineering Environment Enzymes Exhibits Fluorescence Microscopy Genetic Goals Guanosine Triphosphate Phosphohydrolases Human Immune Immune response Immune system Immunologic Receptors Immunomodulators In Vitro Infection Inflammation Innate Immune System Invaded Knowledge Leukocytes Lipid Bilayers Lipids Malignant Neoplasms Membrane Molecular Monomeric GTP-Binding Proteins Movement Pathogenicity Phenotype Phosphatidylinositol Phosphates Phosphatidylinositols Phosphatidylserines Phosphoric Monoester Hydrolases Plasma Proteins Receptor Activation Regulation Reporting Research Research Personnel Role Scaffolding Protein Signal Transduction Signaling Molecule Source System Techniques Technology Testing Travel Virus base cancer cell cell motility chemical release combat fMet-Leu-Phe receptor fluorescence imaging genome editing inhibitor/antagonist inorganic phosphate live cell imaging molecular imaging mutant neutrophil novel therapeutics optogenetics pathogen phosphoinositide-3,4,5-triphosphate phosphoinositide-3,4-bisphosphate reconstitution recruit scaffold single molecule tool

项目摘要

Research Summary Statement Immune cells interpret chemical cues in their environment and make decisions that control their fate. For example, human neutrophils often respond to signals by polarizing and migrating toward the chemical source. Critical for these cellular functions is the dynamic interplay between cell surface receptors, small GTPases, and the enzymes that synthesize phosphatidylinositol phosphate (PIP) lipids. This study aims to decipher the mechanisms the control communication between these different classes of signaling molecules at the plasma membrane. Using human neutrophils, we find that Cdc42 GTPase and a lipid phosphatase denoted SHIP1 trigger a molecular signal that propagates across the plasma membrane in the form of a traveling wave. This behavior, coined excitability, involves repetitive cycles of protein recruitment ON and OFF the membrane. The goal of this study is to determine how SHIP1 regulates the excitable signaling network by integrating signals derived from lipids and membrane tethered proteins. Using a variety of in vitro biochemistry techniques, including supported membrane technology and single molecule imaging, we will determine how lipid composition controls SHIP1 membrane association and phosphatase activity (Aim 1). Using factors that regulate the excitable signaling network in cells, we will reconstitute mechanisms that control SHIP1 membrane recruitment, release of autoinhibition, and activation (Aim 2). In parallel, we will use CRISPR based genome editing, optogenetics, and quantitative live cell imaging of fluorescent biosensors to elucidate how communication between PIP lipids and small GTPase is regulated by SHIP1. Using these tools we will determine the role SHIP1 serves as signaling network scaffold versus a lipid phosphatase (Aim 3). Overall, this study will unify membrane biophysics and cell biology to explain how PIP lipids, small GTPases, and SHIP1 synergistically control the excitable signaling network and cell migration in neutrophils. By unraveling how white blood cells sense, interpret, and respond to pathogenic signals we will fill a gap in knowledge concerning how these signaling molecules coordinate cellular movement with the underlying excitable network. This discovery could open doors for researchers to develop new therapeutics that can be used to modulate immune cell functions in ways that combat infection, inflammation, and cancer.

研究摘要声明免疫细胞在其环境中解释化学提示，并做出控制其命运的决定。为了例如，人类嗜中性粒细胞通常通过对化学源偏振和迁移来对信号做出反应。这些细胞功能至关重要的是细胞表面受体，小GTPase和合成磷脂酰肌醇（PIP）脂质的酶。这项研究旨在破译机制在等离子体上这些不同类别的信号分子之间的控制通信膜。使用人类嗜中性粒细胞，我们发现Cdc42 GTPase和脂质磷酸酶表示为Ship1 触发一个分子信号，该信号以行动波的形式在质膜上传播。这行为，引起的兴奋性涉及在膜上和膜上的蛋白质募集的重复循环。这这项研究的目标是确定Ship1如何通过集成信号来调节可兴奋的信号网络源自脂质和膜拴系蛋白。使用各种体外生物化学技术，包括支持的膜技术和单分子成像，我们将确定脂质组成如何控制 Ship1膜关联和磷酸酶活性（AIM 1）。使用调节兴奋的因素电池中的信号网络，我们将重建控制Ship1膜募集的机制，释放自身抑制和激活（AIM 2）。同时，我们将使用基于CRISPR的基因组编辑，光遗传学，和荧光生物传感器的定量活细胞成像，以阐明PIP脂质之间的通信方式小型GTPase由Ship1调节。使用这些工具，我们将确定Ship1的角色作为信号网络支架与脂质磷酸酶（AIM 3）。总体而言，这项研究将统一膜生物物理和细胞生物学解释PIP脂质，小GTPases和Ship1如何协同控制可激发的信号中性粒细胞中的网络和细胞迁移。通过揭示白细胞的感知，解释和反应致病信号我们将填补有关这些信号分子如何坐在细胞的知识的空白具有基础兴奋网络的运动。这一发现可能为研究人员开发开发可用于以对抗感染的方式调节免疫细胞功能的新疗法，炎症和癌症。