Optogenetic protein design for hypertrophic signaling pathways

肥大信号通路的光遗传学蛋白质设计

• 批准号: 10641938
• 负责人: Lauren L Haar
• 金额: $ 38.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10641938
• 关键词: Action Potentials; Address; Adenylate Cyclase; Applications Grants; Behavior; Behavior Control; Biochemical; Biochemical Pathway; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Categories; Cell membrane; Cells; Communication; Complex; Coupling; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cytoskeleton; Data; Deposition; Development; Dimensions; Disease; Engineering; Event; Extracellular Matrix; Failure; Fetal Growth; Fibroblasts; Fibrosis; Generations; Genes; Heart; Heart Hypertrophy; Heart failure; Homeostasis; Hypertrophy; Individual; Ion Channel Gating; Light; Location; Lower Organism; Maps; Measures; Mediating; Mitochondria; Myocardial; Nature; Neurons; Neurosciences; Organ; Pathologic; Pathway interactions; Physiological; Play; Population; Production; Proliferating; Protein Engineering; Protein Isoforms; Protein Kinase; Protein Kinase C; Proteins; Research; Resolution; Role; Sarcoplasmic Reticulum; Series; Signal Pathway; Signal Transduction; Signaling Protein; Site; Stimulus; Stress; Technology; Tissue-Specific Gene Expression; Validation; Work; Workload; analog; biological adaptation to stress; cell behavior; cell type; coronary fibrosis; design; extracellular; intercellular communication; interest; light gated; neural circuit; novel; optogenetics; programs; response; spatiotemporal; therapeutic target; tool

Optogenetics has primarily employed light gated ion channels (derived from lower organisms) to trigger or inhibit the neuronal action potential. Analogous studies in the cardiovascular field have used these photoresponsive foreign proteins to regulate contractility and pacing. However, light gated ion channels are unable to interrogate the intracellular mechanisms that control behavior and homeostasis. As a consequence, we have developed a protein engineering strategy for creating light responsive analogs of endogenous proteins. The proposed research program is focused on the design, construction, and validation of optogenetic signaling for proteins that contribute to myocardial homeostasis, and the application of this technology to unravel the mechanisms that drive cardiac hypertrophy and fibrosis. Aim 1: Optogenetic Engineering to Access the Biochemical Pathways of Hypertrophy and Fibrosis. We will employ a novel optogenetic engineering strategy to create light responsive signaling cascades localized at specific intracellular sites, with a focus on cAMP-mediated signaling pathways. Aim 2: Light Guided Mapping of the Signaling Domains that Mediate Hypertrophy. The optogenetic constructs outlined in Aim 1 will be used to explore the impact of signaling events at distinct intracellular locations on cardiomyocyte behavior, with an emphasis on hypertrophic relevant responses. Aim 3: Light Guided Mapping of the Intercellular Communication that Mediates Fibrosis. Optogenetic analogs of endogenous proteins will be used to assess intercellular crosstalk between two distinct cardiac cell types. Light- triggered activation of engineered signaling pathways offers the opportunity to tease apart a collaborative signaling network that has been observed between these distinct cell populations.

光遗传学主要利用光门控离子通道（源自低等生物）来触发或抑制 神经元动作电位。心血管领域的类似研究已经使用了这些光响应 外来蛋白质调节收缩性和起搏。然而，光门控离子通道无法询问 控制行为和体内平衡的细胞内机制。因此，我们开发了一个 用于创建内源蛋白质的光响应类似物的蛋白质工程策略。拟议的 研究项目的重点是蛋白质光遗传学信号的设计、构建和验证， 有助于心肌稳态，并且应用该技术来揭示心肌稳态的机制 促使心脏肥大和纤维化。目标1：光遗传学工程获取生化物质 肥大和纤维化的途径。我们将采用一种新颖的光基因工程策略来创造 光响应信号级联位于特定的细胞内位点，重点是 cAMP 介导的 信号通路。目标 2：介导肥大的信号传导域的光导绘图。这 目标 1 中概述的光遗传学结构将用于探索不同信号事件的影响 细胞内位置对心肌细胞行为的影响，重点是肥厚相关反应。目标 3： 介导纤维化的细胞间通讯的光导绘图。光遗传学类似物 内源蛋白将用于评估两种不同心脏细胞类型之间的细胞间串扰。光- 工程信号通路的触发激活提供了梳理协作的机会 在这些不同的细胞群之间观察到的信号网络。