Phospholipase D Regulation of Exosome Secretion

磷脂酶 D 对外泌体分泌的调节

• 批准号: 2325227
• 负责人: Michelle Knowles
• 金额: $ 63.95万
• 依托单位: University of Denver
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2026-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2325227&HistoricalAwards=false
• 关键词: Phospholipase; Regulation; Exosome; Secretion

With the support of the Chemistry of Life Processes Program in the Division of Chemistry, Michelle Knowles of the University of Denver is studying how cells control their communication with one another. One way that cells communicate is through the secretion of nanoscopic packets called “exosomes.” Exosomes are lipid-coated containers that carry molecules and travel through the body, where they are taken up by other cells. Exosomes can alter the function of other cells, for better or worse. The factors that control how cells make exosomes and their release are not fully understood. The research supported by this award will focus on the lipid-modifying protein, phospholipase D (PLD), an enzyme, and on phosphatidic acid (PA), which is a lipid whose production is catalyzed by PLD. PA can alter the shapes of membranes and acts as a signaling molecule. The role of PLD and PA in the formation of exosomes and their release will be investigated. Beyond generating new knowledge about exosomes, the project will create opportunities for graduate and undergraduate students to learn how to use state of the art technology to address problems at the forefront of cell communication. Drs. Knowles and Dinah Loerke (University of Denver) will develop a peer mentoring program for graduate students. The results of scientific and broader impact activities will be presented at conferences and published to allow others to build upon the foundational knowledge obtained about cellular regulation and student mentorship.Phospholipase D and phospholipase A (PLD and PA) could be involved in the formation of exosomes and/or in the release process itself. Both steps require highly curved membranes. In this research project, the presence of PLD and the formation of PA will be mapped in space and time during exosome secretion. Quantitative fluorescence microscopy approaches will be used in conjunction with standard biochemical methods to assess the number of exosomes and the frequency with which they are released from cells. The formation, trafficking and fusion stages of the exosome lifecycle will be assessed in live cells. The activity and presence of PLD will be altered to determine when and where PLD and phosphatidic acid are required. This project has the potential for far-reaching scientific impact; specifically, by demonstrating the use of biophysical tools/chemical biology to help to elucidate important mechanisms of cellular communication.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学过程中的化学过程中的支持下，丹佛大学的米歇尔·诺尔斯（Michelle Knowles）正在研究细胞如何控制彼此的交流。细胞传达的一种方法是通过称为“外泌体”的纳米镜包的分泌。外泌体是载有分子并穿过体内的脂质涂层容器，在那里它们被其他细胞吸收。外泌体可以改变其他细胞的功能，无论好坏。控制细胞如何使外泌体及其释放的因素尚未完全理解。该奖项支持的研究将重点放在脂质修饰的蛋白质，磷脂酶D（PLD），一种酶和磷脂酸（PA）上，磷脂酸（PA）是一种脂质，其生产由PLD催化。 PA可以改变膜的形状，并用作信号分子。 PLD和PA在外泌体形成及其释放中的作用将得到研究。除了产生有关外泌体的新知识外，该项目还将为研究生和本科生创造机会，以学习如何利用最先进的技术来解决细胞通信最前沿的问题。博士。 Knowles和Dinah Loerke（丹佛大学）将为研究生制定同伴心理计划。科学和更广泛的影响活动的结果将在会议上呈现，并发表以允许其他人建立在有关细胞调节和学生心态的基础知识的基础上。磷脂酶D和磷脂酶A（PLD和PA）可以参与外泌体的形成和/或在释放过程中。这两个步骤都需要高度弯曲的机制。在该研究项目中，PLD的存在和PA的形成将在外泌体分泌过程中的空间和时间绘制。定量荧光显微镜方法将与标准生化方法结合使用，以评估外泌体的数量及其从细胞中释放的频率。外泌体生命周期的形成，运输和融合阶段将在活细胞中评估。将改变PLD的活性和存在，以确定需要何时何地需要PLD和磷脂酸。该项目有可能产生深远的科学影响。具体而言，通过证明使用生物物理工具/化学生物学来帮助阐明细胞通信的重要机制。该奖项反映了NSF的法定任务，并通过使用基金会的知识分子优点和更广泛的影响评估标准来通过评估来表现出珍贵的支持。