Molecular identity of exosomal BK channels

外泌体 BK 通道的分子特性

• 批准号: 10366418
• 负责人: Mahmood Khan
• 金额: $ 62.62万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-24 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10366418
• 关键词: Animals; Attention; Biochemical; Biogenesis; Biological; Brain Hypoxia-Ischemia; Calcium; Cardiac; Cardiac Myocytes; Cell Death; Cell membrane; Cells; Cytoplasm; Data; Data Analyses; Disease; Disease Marker; Distant; Docking; Drug Delivery Systems; Electrophysiology (science); Environment; Face; Future; Genetic; Goals; Heart; Homeostasis; Human; Investigation; Ion Channel; Ion Exchange; Ions; Knockout Mice; Link; Liposomes; Measures; Mediator of activation protein; Medicine; Membrane; Messenger RNA; Methods; MicroRNAs; Microscopy; Mitochondria; Molecular; Molecular Biology; Molecular Target; Mus; Myocardial Infarction; Myocardial Ischemia; Organelles; Osmolar Concentration; Osmotic Shocks; Outcome; Physiological; Physiology; Plasma; Play; Potassium; Potassium Channel; Process; Proteins; Proteomics; Pump; RNA Splicing; Reperfusion Injury; Reporting; Resolution; Role; Signal Transduction; Source; Stress; Techniques; Testing; Therapeutic; Time; Tissues; Variant; Voltage-Gated Potassium Channel; Wild Type Mouse; biophysical properties; body system; cardioprotection; coronary fibrosis; delivery vehicle; exosome; experimental study; extracellular; extracellular vesicles; gain of function; imaging approach; in silico; induced pluripotent stem cell derived cardiomyocytes; innovation; intercellular communication; large-conductance calcium-activated potassium channels; macromolecule; molecular imaging; mouse model; mutant; new technology; novel; patch clamp; programs; sensor; targeted delivery; therapeutic development; therapeutic target; transmission process

Abstract Extracellular vesicles (EVs) have gained significant attention since their discovery in 1983 as important mediators of intercellular communications, potential disease markers, therapeutic targets, and drug delivery vehicles. Though it is widely accepted that EVs get packaged inside the cell, pass through the extracellular environment, and deliver the cargo to the target cells. However, even after 37 yrs it is not determined, 1) how EVs handle the differential ionic environment (cytoplasm vs extracellular), 2) whether EVs possess any functional ion channels, and 3) whether any of these channels play a physiological role. We focused on answering these questions and focused on an ion with the largest gradient, i.e., potassium. Using the in silico approach, we discovered several ion channels, and the most prominent ion channels, we discovered in exosomes is BK. We incorporated a novel electrophysiology approach, near field electrophysiology, as canonical patch-clamp methods are not feasible due to the size of exosomes. We discovered that functional BK channels exist in exosomes, and decide the integrity of exosomes. Our preliminary data also indicate that exosomal BK can protect the heart from ischemia-reperfusion injury. We will now test the hypothesis that exosomes containing BK determine the content of exosomes, facilitate their survival in variable ionic environments, and protect the heart from IR injury. Overall the data supports the above hypothesis which will be tested using multiple approaches and pursuing the following specific aims to, 1. establish a presence, molecular identity, and biophysical properties of BK in exosomes, 2. determine the physiological role of BK in exosomes., and 3. elucidate the mechanistic role of exosomal BK channels in cardioprotection. In our proposal, we have incorporated genetic mice models, and innovative as well as a novel technology to understand a very basic and broad biological question. The outcome of this program will open an opportunity to study exosomal ion channels including BK channels, and advance the exosome field by determining how exosome survive variable osmolarities, establishing the molecular identity of exosomal ion channels, understand how cargo content is regulated by exosomal ion channels, and the role and mechanism of exosomal ion channels in cardioprotection. In the future, our study will set the ground for exploring other ion channels in exosomes from different living beings as well as organ systems.

摘要 自1983年发现细胞外囊泡以来， 细胞间通讯的介质、潜在的疾病标志物、治疗靶点和药物 送货车辆。虽然人们普遍认为电动汽车被包装在电池内，但通过电池， 细胞外环境，并将货物递送至靶细胞。然而，即使在37岁之后， 确定，1）EV如何处理不同的离子环境（细胞质与细胞外），2）EV是否 具有任何功能性离子通道，以及3）这些通道中的任何一个是否发挥生理作用。我们 专注于回答这些问题并专注于具有最大梯度的离子，即，钾使用 在计算机模拟的方法中，我们发现了几个离子通道，最突出的离子通道，我们 我们采用了一种新的电生理学方法，近场 由于外来体的大小，典型的膜片钳方法是不可行的。我们 发现功能性BK通道存在于exosomes中，并决定exosomes的完整性。我们 初步数据还表明外泌体BK可以保护心脏免受缺血-再灌注损伤。我们 现在将测试含有BK的外泌体决定外泌体的含量的假设， 它们在可变离子环境中的存活，并保护心脏免受IR损伤。总体而言，数据支持 上述假设将使用多种方法进行测试，并追求以下具体目标， 1.建立BK在外来体中的存在、分子身份和生物物理性质，2. 确定BK在外泌体中的生理作用，和3.阐明外泌体的机制作用 BK通道在心脏保护中的作用。在我们的提案中，我们纳入了遗传小鼠模型， 这是一种创新的新技术，可以理解一个非常基本和广泛的生物学问题。 该计划的结果将为研究包括BK在内的外泌体离子通道提供机会 通道，并通过确定外泌体如何存活变量来推进外泌体领域 渗透压，建立外泌体离子通道的分子身份，了解货物含量 是由外泌体离子通道调节的，外泌体离子通道在 心脏保护在未来，我们的研究将为探索外泌体中的其他离子通道奠定基础， 不同的生物以及器官系统。