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

• 批准号: 10388807
• 负责人: Guolin Ma
• 金额: $ 22.57万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 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/10388807
• 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; Pharmacology; Photosensitivity; 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; base; biological systems; design; endoplasmic reticulum stress; extracellular; genetic manipulation; human disease; immunoregulation; in vivo; innovation; nervous system disorder; neuroregulation; novel; novel strategies; optogenetics; peroxisome; protein folding; remote control; response; spatiotemporal; technology development; tool; ultraviolet irradiation; voltage

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.

肌肉细胞中的内质网（ER）或肌质网，是最大的细胞内细胞内的 Ca2+ Store并在塑造Ca2+ Sigals的时空动力学方面起关键作用 细胞内Ca2+体内平衡。相互地，钙也与ER功能密切相关， 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的两侧的Ca2+浓度 通过膜接触部位与周围的细胞器进行介导Ca2+转移。放松管制的ER Ca2+ 稳态不仅会导致ER胁迫和展开的蛋白质反应（UPR），而且还参与 心血管疾病，神经系统疾病，代谢综合征和其他疾病。 药理调节和遗传操作通常用于研究ER CA2+处理 机械，但是这些基本上不可逆的方法通常缺乏空间精度和特异性。 光遗传学技术提供了一种新颖的方法，用于调节ER CA2+稳态 高级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，称为遗传编码的ER钙致动剂（Geecas）， 启用ER CA2+稳态，ER-Organelle CA2+通信和细胞器的光学询问 CA2+调制活性在多个生物系统中。在特定的目标1中，团队将设计一组 基于ER定位的钙通道和/或其照片传动器的地理位置 ER Ca2+信号具有不同的动力学和动态特性。在特定的目标2中，团队试图发展 一个光遗传平台，用于控制ER及其之间的Organellar束缚和Ca2+转移 周围的细胞器。该项目的成功执行将为 实现ER Ca2+体内稳态和孔间CA2+信号传导的无创和精确调制 具有高时空精度，从而对细胞生理进行遥控器，包括ER 应力，能量代谢，自噬和MTOR信号传导。从转化的角度，分子 该项目产生的工具将为人类疾病提供新颖的介入方法 与异常的ER Ca2+信号传导相关。