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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），或肌细胞中的肌浆网，充当最大的细胞内结构 Ca2+储存并在塑造Ca2+信号的时空动态中起着关键作用，细胞内Ca2+稳态。反过来，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采用一系列调节剂来控制膜两侧的Ca 2+浓度，介导的Ca2+转移与周围的细胞器通过膜接触网站。去调节ER Ca2 + 稳态不仅导致内质网应激和未折叠蛋白反应（UPR），而且还参与了心血管疾病、神经系统疾病、代谢综合征和其他疾病。药物调节和遗传操作常被用于研究ER Ca 2+处理这些基本上不可逆转的方法往往缺乏空间精确性和针对性。光遗传学技术提供了一种新的方法来调节ER Ca 2+稳态，上级时空分辨率和高可逆性。因此，该团队建议创建一个创新的光遗传学工具包，命名为遗传编码的ER钙致动器（GEECA），能够对ER Ca2+稳态、ER-细胞器Ca2+通讯和细胞器多种生物系统中的Ca2+调节活性。在具体目标1中，团队将设计一套基于ER定位的钙通道和/或其光可切换致动器的GEECA以进行光调谐 ER Ca2+信号具有不同的动力学和动态特性。在具体目标2中，该团队寻求开发一个光遗传学平台，以控制细胞器间的束缚和钙离子转移之间的ER和其周围的细胞器。该项目的成功实施将为以下方面提供新的机会：实现ER Ca 2+稳态和细胞器间Ca 2+信号传导的非侵入性和精确调节具有高时空精度，从而对包括ER在内的细胞生理学施加远程控制应激、能量代谢、自噬和mTOR信号传导。从翻译的角度来看，该项目产生的工具将为人类疾病提供新的干预方法与异常ER Ca2+信号传导相关。