High-throughput integrated live imaging and optogenetic pacing platform to assess hypoxia responsiveness in the fly heart

高通量集成实时成像和光遗传学起搏平台，用于评估果蝇心脏的缺氧反应

• 批准号: 10132500
• 负责人: Chao Zhou
• 金额: $ 58.67万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10132500
• 关键词: Abdomen; Aging; Anatomy; Animal Model; Autophagocytosis; Behavioral; Biological Assay; Biological Models; Bradycardia; Cardiac; Cardiac Function Study; Collaborations; Color; Consumption; Development; Dorsal; Drosophila genus; Drosophila melanogaster; Drug Targeting; Electric Stimulation; Future; Gene Proteins; General Hospitals; Genes; Genetic; Genetic Models; Glean; Heart; Heart Arrest; Heart Diseases; Heart Injuries; Heart Rate; Human; Hypoxia; Image; Imaging technology; Intelligence; Ion Channel; Ischemia; Ischemic Preconditioning; Knowledge; Light; Lysosomes; Massachusetts; Modeling; Molecular; Morphologic artifacts; Names; Opsin; Optical Coherence Tomography; Optics; Oranges; Organism; Pathway interactions; Physiology; Prevention; Publishing; Recovery; Reperfusion Therapy; Research; Research Design; Role; Science; Series; Side; Stress; Structure; Surface; System; Tachycardia; Technology; Testing; Time; Tissues; Transgenic Organisms; Tube; Universities; Washington; base; cardioprotection; deep learning; design; experimental study; fly; heart function; heart imaging; heart rhythm; human disease; image processing; in vivo; insight; instrument; non-invasive imaging; novel; optical imaging; optogenetics; preconditioning; programs; real-time images; relating to nervous system; response; segmentation algorithm; tool

Project Summary Ischemic preconditioning is a well-established phenomenon, in which a brief episode(s) of controlled ischemia and reperfusion renders cardioprotection from a subsequent sustained episode of ischemia. An emerging body of evidence demonstrated that neural regulated heart rate modulation confers cardiac preconditioning responses. Understanding the mechanism through model systems of preconditioning would help us identify the genes and proteins when designing future drug targets for the prevention of ischemic cardiac injury. As a promising alternative to electrical pacing to modulate heart rate, optogenetic pacing does not require physical contact, has high spatial and temporal precision, offers more specific excitation, and avoids artifacts from electrical stimulation. Recent developments in the field of optogenetics make it possible for non-invasive and specific optical control of the heart rhythm in animal models, such as in Drosophila melanogaster. Drosophila is a powerful genetic model system that has been used since the early 1900s to characterize genes associated with human diseases, including cardiac diseases. Studies performed in flies can provide insights into conserved mechanisms in cardiac diseases, which can be applied to higher organisms, including humans. Working in collaboration with Drs. Airong Li and Rudolph Tanzi from the Massachusetts General Hospital, we demonstrated non-invasive optogenetic pacing and concurrent optical coherence tomography (OCT) imaging of the Drosophila heart for the first time. Recently, we further demonstrated red-light optogenetic pacing and successful optical control of tachycardia, bradycardia, and restorable cardiac arrest in fly models. Building on the decade-long productive collaboration with Drs. Li and Tanzi and new collaborations with Dr. Abhinav Diwan (cardiologist) and Dr. Jeanne Nerbonne (cardiac electrophysiologist) and Dr. Kenneth Schechtman (biostatistician) at Washington University, we propose to develop a high-throughput integrated OCT imaging and dual-color optogenetic pacing system and establish a novel research platform to study preconditioning and hypoxia responsiveness in the fly heart. We hypothesize that periods of bradypacing will precondition the fly heart to protect against hypoxia, via activation of the autophagy-lysosome pathway. The specific aims are: 1) Develop and optimize a high-throughput integrated instrument for non-invasive OCT imaging and optogenetic control of fly heart function in vivo; 2) Develop double transgenic fly models and functional assays based on OCT imaging to characterize fly heart physiology in vivo; 3) Define functional and molecular changes in response to hypoxia and optogenetic preconditioning in transgenic fly models. If successful, the high-throughput optical imaging and dual-color optogenetic pacing platform developed in this program combined with powerful double transgenic fly models will enable us to characterize changes of the fly heart function in response to different stress challenges that is not feasible before. This will allow us to perform a series of new experiments, providing insights into conserved molecular mechanisms on hypoxia-induced cardiac changes and preconditioning.

项目摘要 缺血性预处理是一种完善的现象，其中简短的情节受控缺血 再灌注可从随后持续的缺血发作中产生心脏保护。一个新兴的身体 证据表明，神经调节的心率调节赋予心脏预处理 回答。通过预处理模型系统了解机制将有助于我们确定 设计未来的药物靶标在预防缺血性心脏损伤时，基因和蛋白质。作为 有望替代电动起搏以调节心率，光遗传起搏不需要物理 接触，具有较高的空间和时间精度，提供了更具体的激发，并避免了 电刺激。光遗传学领域的最新发展使得无创和 动物模型中心脏节奏的特定光学控制，例如在果蝇中。果蝇是 自1900年代初以来一直使用的强大遗传模型系统来表征相关的基因 与人类疾病，包括心脏疾病。用苍蝇进行的研究可以提供对保守的见解 心脏病的机制，可以应用于包括人类在内的更高生物体。工作 与Drs合作。来自马萨诸塞州综合医院的Airong Li和Rudolph Tanzi，我们证明了 果蝇的非侵入性光学起搏和并发光相干断层扫描（OCT）成像 第一次心。最近，我们进一步证明了红光光学遗传节奏和成功的光学 在飞行模型中控制心动过速，心动过缓和可修复的心脏骤停。在长达十年 与Drs的生产合作。 Li和Tanzi以及与Abhinav Diwan博士（心脏病专家）的新合作 以及Jeanne Nerbonne博士（心脏电生理学家）和Kenneth Schechtman博士（生物统计学家） 华盛顿大学，我们建议开发高通量集成的OCT成像和双色 光遗传进攻系统并建立了一个新的研究平台来研究预处理和缺氧 飞翔的心脏反应。我们假设勃ardypac的时期将使苍蝇心脏前提 通过激活自噬溶质体途径来预防缺氧。具体目的是：1）发展 并优化一种用于非侵入性OCT成像和光遗传控制的高通量集成仪器 在体内飞行心脏功能； 2）基于OCT成像开发双转基因飞行模型和功能测定 在体内表征苍蝇心生理； 3）定义功能和分子变化，以响应缺氧 和转基因蝇模型中的光遗传学预处理。如果成功，高通量光学成像和 该程序中开发的双色光遗传步调平台与强大的双转基因苍蝇相结合 模型将使我们能够以不同的压力挑战来表征苍蝇心脏功能的变化 这是不可行的。这将使我们能够执行一系列新实验，从而提供有关 缺氧引起的心脏变化和预处理的保守分子机制。