Optogenetic toolkit for precise control of organellar calcium signaling

用于精确控制细胞器钙信号传导的光遗传学工具包

基本信息

批准号：
10706462
负责人：
Yubin Zhou
金额：
$ 18.78万
依托单位：
TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
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：
2022-09-20 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10706462
关键词：
Autophagocytosis Biomedical Research Calcium Calcium Channel Calcium Signaling Cardiovascular Diseases Cell Surface Receptors Cell membrane Cell physiology Cellular biology Communication Coupling Cytoplasm Diglycerides Disease Drosophila melanogaster Endoplasmic Reticulum Energy Metabolism Engineering FRAP1 gene G-Protein-Coupled Receptors Generations Goals Homeostasis Intervention Kinetics Light Lipids Lysosomes Malignant Neoplasms Mammalian Cell Mediating Membrane Metabolic syndrome Mitochondria Modification Molecular Muscle Cells Names Neurodegenerative Disorders Optics Organelles Outer Mitochondrial Membrane Oxygen Pathway interactions Pattern Performance Phototoxicity Physiological Processes Play Positioning Attribute Precision therapeutics Property Protein Biosynthesis Protein Engineering Proteins Receptor Protein-Tyrosine Kinases Resolution Role STIM1 gene Sarcoplasmic Reticulum Series Shapes Side Signal Transduction Signaling Molecule Site Specificity System Technology Testing Toxic effect biological systems design endoplasmic reticulum stress extracellular genetic manipulation human disease immunoregulation in vivo innovation invention nervous system disorder neuroregulation novel novel strategies optogenetics peroxisome pharmacologic protein folding release of sequestered calcium ion into cytoplasm remote control response spatiotemporal technology development tool ultraviolet irradiation voltage

项目摘要

Endoplasmic reticulum (ER), or sarcoplasmic reticulum in muscle cells, acts as the largest intracellular Ca2+ store and plays a pivotal role in shaping the spatiotemporal dynamics of Ca2+ sigals to maintain intracellular Ca2+ homeostasis. Reciprocally, calcium is also intimately involved in regulating ER functions, including lipid synthesis, protein synthesis, folding, modifications and translocation. Consequently, the ER employs a series of regulators to control Ca2+ concentration on both sides of the membrane and mediate Ca2+ transfer with the surrounding organelles via membrane contact sites. Deregulated ER Ca2+ homeostasis not only results in ER stress and unfolded protein response (UPR), but also is involved in cardiovascular diseases, neurological diseases, metabolic syndromes and other diseases. Pharmacological modulation and genetic manipulations are often applied to study the ER Ca2+ handling machinery, but these largely irreversible approaches often lack spatial precision and specificity. Optogenetics technology provides a novel approach for modulating ER Ca2+ homeostasis with superior spatiotemporal resolution and high reversibility. Hence, the team proposes to create an innovative optogenetic toolkit, named as Genetically Encoded ER Calcium Actuators (GEECAs), that enable optical interrogation of ER Ca2+ homeostasis, ER-organelle Ca2+ communications and organellar Ca2+-modulated activities in multiple biological systems. In Specific Aim 1, the team will design a set of GEECAs based on ER-localized calcium channels and/or their photoswitchable actuators to photo-tune ER Ca2+ signals with varying kinetic and dynamic properties. In Specific Aim 2, the team seeks to develop an optogenetic platform to control inter-organellar tethering and Ca2+ transfer between ER and its surrounding organelles. The successful execution of this project will provide novel opportunities to achieve noninvasive and precise modulation of ER Ca2+ homeostasis and inter-organellar Ca2+ signaling with high spatiotemporal precision, thereby exerting remote control over cellular physiology, including ER stress, energy metabolism, autophagy and mTOR signaling. From a translational perspective, molecular tools generated from this project will provide novel interventional approaches for human diseases associated with aberrant ER Ca2+ signaling.

内质网 (ER) 或肌肉细胞中的肌浆网，充当最大的细胞内网 Ca2+ 储存并在塑造 Ca2+ 信号的时空动态以维持细胞内 Ca2+ 稳态。相反，钙也密切参与调节 ER 功能，包括脂质合成、蛋白质合成、折叠、修饰和易位。因此， ER 采用一系列调节器来控制膜两侧的 Ca2+ 浓度，通过膜接触位点介导 Ca2+ 与周围细胞器的转移。 ER Ca2+ 解除管制稳态不仅导致 ER 应激和未折叠蛋白反应 (UPR)，而且还参与心血管疾病、神经系统疾病、代谢综合征等疾病。药理学调节和基因操作通常用于研究 ER Ca2+ 处理机械，但这些基本上不可逆的方法往往缺乏空间精度和特异性。光遗传学技术提供了一种调节 ER Ca2+ 稳态的新方法优越的时空分辨率和高可逆性。因此，团队建议创建一个创新的光遗传学工具包，称为基因编码内质网钙执行器（GEECA），实现内质网 Ca2+ 稳态、内质网细胞器 Ca2+ 通讯和细胞器的光学询问多种生物系统中 Ca2+ 调节的活性。在具体目标 1 中，团队将设计一组基于内质网局部钙通道和/或其光开关致动器进行光调谐的 GEECA ER Ca2+ 信号具有不同的动力学和动态特性。在具体目标 2 中，团队力求开发一个光遗传学平台来控制内质网与其之间的细胞器间束缚和 Ca2+ 转移周围的细胞器。该项目的成功实施将为实现内质网 Ca2+ 稳态和细胞器间 Ca2+ 信号传导的无创和精确调节具有高时空精度，从而对细胞生理学进行远程控制，包括 ER 应激、能量代谢、自噬和 mTOR 信号传导。从翻译的角度来看，分子该项目产生的工具将为人类疾病提供新的介入方法与异常的 ER Ca2+ 信号传导有关。