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Optogenetic stimulation of TMEM16F to control phospholipid flip-flop

TMEM16F 的光遗传学刺激控制磷脂触发器

基本信息

批准号：
10601109
负责人：
Huanghe Yang
金额：
$ 23.38万
依托单位：
DUKE UNIVERSITY
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：
2022-04-05 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10601109
关键词：
Anemia Animal Model Apoptosis Biological Assay Biological Process Biology Biomedical Engineering Biophysics Blood coagulation Bypass C-terminal COVID-19 Calcium Caspase Cell fusion Cell membrane Cells Cellular biology Chemicals Coupled Coupling Detection Disease Engineering Environment Fertilization G-Protein-Coupled Receptors Genetic Goals HIV Health Hematology Immunology In Vitro Ion Channel Ions Light Lipids Location Malignant Neoplasms Medicine Membrane Membrane Biology Modernization Molecular Molecular Conformation Myocardial Infarction Names Oxygen Pathologic Processes Permeability Phagocytosis Phospholipids Physiologic calcification Physiological Physiological Processes Physiology Pregnancy Complications Process Property Protein Engineering Proteins Protocols documentation Research Resolution SARS-CoV-2 infection Signal Transduction Specificity Stroke Structure Time Translating Vesicle Virus Diseases cell motility cell type experience feasibility testing fluorescence imaging genetic manipulation high reward high risk in vivo Model novel optical imaging optogenetics passive transport pharmacologic phospholipid scramblase prototype repaired response spatiotemporal tool vesicular release virology voltage

项目摘要

SUMMARY Phospholipid flip-flop on cell membranes can exert profound impacts on cellular signaling and functions, including apoptosis, phagocytosis, blood coagulation, membrane vesicle shedding, bone mineralization, cell-cell fusion, fertilization, viral infection including HIV and SARS-CoV2 infections. Nevertheless, how phospholipid flip-flop leads to the observed cellular responses is largely elusive. The recent identifications of phospholipid scramblases and flippases have enabled genetic manipulations of these critical phospholipid transporters, which greatly advanced our understanding on the biology of phospholipid flip-flop. However, phospholipid flip-flop is a dynamic process and the genetic manipulations only allow us to observe the end results, which hinders gaining mechanistic understanding phospholipid flip-flop in various physiological process in real time. In this application, we aim to test the feasibility of developing a genetically encoded, optogenetic toolbox to precisely control phospholipid flip-flop with light at high temporal and spatial resolution and in real time. Our proposal is based on our extensive experience on the recently discovered calcium-activated phospholipid scramblase (CaPLSase) TMEM16F at molecular and cellular levels. In response to intracellular calcium increase, TMEM16F can rapidly catalyze phospholipid flip-flop, efficiently disrupt membrane environment and trigger wide spectrum of cellular changes. Here, we will use two complementary but independent approaches to develop the optogenetic tools to control phospholipid flip-flop. First, we will coexpress various calcium- mobilizing optogenetic tools to indirectly activate TMEM16F utilizing its calcium sensing property. Second, we will engineer light sensing motifs into TMEM16F to enable direct light control of its activities. If successful, the optogenetic toolbox developed in this high-risk, high-reward application will have profound impacts and broad applications in membrane biology, cell biology, physiology, hematology, immunology, virology and medicine.

总结