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: 10318214
• 负责人: Chao Zhou
• 金额: $ 54.73万
• 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/10318214
• 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; 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.

项目概要 缺血预适应是一种公认​​的现象，其中受控缺血的短暂发作 再灌注可以保护心脏免受随后持续的缺血发作的影响。新兴机构 大量证据表明，神经调节心率调节可实现心脏预适应 回应。通过预处理模型系统了解该机制将有助于我们识别 在设计未来预防缺血性心脏损伤的药物靶标时，需要考虑基因和蛋白质。作为一个 光遗传学起搏是替代电起搏来调节心率的有希望的替代方案，不需要物理 接触，具有高空间和时间精度，提供更具体的激励，并避免伪影 电刺激。光遗传学领域的最新发展使得非侵入性和 在动物模型中，例如果蝇，对心律进行特定的光学控制。果蝇是 一个强大的遗传模型系统，自 1900 年代初以来一直被用来描述相关基因的特征 与人类疾病有关，包括心脏病。在果蝇中进行的研究可以提供对保守性的见解 心脏病的机制，可以应用于包括人类在内的高等生物。工作于 与博士的合作。来自麻省总医院的 Airong Li 和 Rudolph Tanzi，我们展示了 果蝇的非侵入性光遗传学起搏和同步光学相干断层扫描 (OCT) 成像 第一次心动。最近，我们进一步展示了红光光遗传学起搏和成功的光学 控制苍蝇模型中的心动过速、心动过缓和可恢复的心脏骤停。建立在长达十年的基础上 与博士的富有成效的合作。 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，我们建议开发一种高通量集成OCT成像和双色 光遗传学起搏系统并建立一个新的研究平台来研究预处理和缺氧 苍蝇心中的反应。我们假设，一段时间的缓慢起搏将使果蝇心脏先于 通过激活自噬-溶酶体途径防止缺氧。具体目标是： 1) 发展 并优化了用于非侵入性 OCT 成像和光遗传学控制的高通量集成仪器 蝇体内心脏功能； 2）开发基于OCT成像的双转基因果蝇模型和功能测定 表征果蝇心脏的体内生理学特征； 3) 定义响应缺氧的功能和分子变化 以及转基因果蝇模型中的光遗传学预处理。如果成功，高通量光学成像和 本项目开发的双色光遗传学起搏平台结合强大的双转基因果蝇 模型将使我们能够表征果蝇心脏功能因应对不同压力挑战而发生的变化 这在以前是不可行的。这将使我们能够进行一系列新的实验，从而深入了解 缺氧引起的心脏变化和预处理的保守分子机制。