Molecular identity of exosomal BK channels

外泌体 BK 通道的分子特性

基本信息

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
项目类别：
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

项目摘要

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.

抽象的细胞外囊泡（EV）自 1983 年被发现以来就受到了广泛的关注，因为它的重要性细胞间通讯介质、潜在疾病标志物、治疗靶点和药物送货车辆。尽管人们普遍认为电动汽车被封装在电池内，但通过细胞外环境，并将货物递送至靶细胞。然而，即使过了37岁，它也没有确定，1) EV 如何处理不同的离子环境（细胞质与细胞外），2) EV 是否是否具有任何功能性离子通道，以及 3) 这些通道是否发挥生理作用。我们专注于回答这些问题并关注梯度最大的离子，即钾。使用通过计算机方法，我们发现了几个离子通道，以及最突出的离子通道，我们在外泌体中发现的是BK。我们采用了一种新颖的电生理学方法，近场电生理学，因为由于外泌体的大小，规范的膜片钳方法不可行。我们发现外泌体中存在功能性BK通道，并决定外泌体的完整性。我们的初步数据还表明，外泌体BK可以保护心脏免受缺血再灌注损伤。我们现在将检验含有 BK 的外泌体决定外泌体含量的假设，促进它们在变化的离子环境中生存，并保护心脏免受红外线损伤。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. 确定外泌体中 BK 的存在、分子身份和生物物理特性，2. 确定 BK 在外泌体中的生理作用。 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，并且创新和新颖的技术来理解一个非常基本和广泛的生物学问题。该计划的成果将为研究外泌体离子通道（包括 BK）提供机会通道，并通过确定外泌体如何生存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特性，了解货物含量受外泌体离子通道的调控，外泌体离子通道的作用和机制心脏保护。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。