Supplement to Elucidating biogenesis and cargo sorting mechanisms for discrete extracellular vesicle subpopulations in C. elegans

补充阐明线虫离散细胞外囊泡亚群的生物发生和货物分选机制

• 批准号: 10643364
• 负责人: Jessica E Tanis
• 金额: $ 5.77万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10643364
• 关键词: Afferent Neurons; Animal Behavior; Animals; Applications Grants; Area; Arts; Biochemistry; Biogenesis; Biological Sciences; Biology; COVID-19 pandemic; Caenorhabditis elegans; Calcium; Candidate Disease Gene; Cell membrane; Cells; Cellular Stress; Chemistry; Chimeric Proteins; Cilia; Communication; Communities; Computer software; Cultured Cells; Data; Defect; Delaware; Development; Educational process of instructing; Endocytosis; Environment; Equipment; Exclusion; Funding; Future; Genetic; Goals; Hermaphroditism; Heterogeneity; Homeostasis; Image; Image Analysis; Individual; Inosine Diphosphate; International; Ion Channel; Kinesin; Label; Lateral; Leadership; Learning; Lipids; Manuscripts; Mass Spectrum Analysis; Measures; Membrane; Mentors; Microscopy; Modeling; Molecular; Monomeric GTP-Binding Proteins; Multivesicular Body; Neurodegenerative Disorders; Neuroglia; Neurons; Neurosciences; Pathologic; Phospholipids; Physiological; Physiological Processes; Plasma; Play; Population; Population Heterogeneity; Proteins; RNA; Research; Resolution; Role; Running; Schools; Site; Sorting - Cell Movement; Stimulus; Structure; System; Technical Expertise; Techniques; Testing; Time; Tissues; Total Internal Reflection Fluorescent; Training; Transgenic Animals; Transgenic Organisms; Transmission Electron Microscopy; United States National Institutes of Health; Universities; Vesicle; Visualization; Writing; animal imaging; base; bioimaging; career; career development; career networking; cell type; exosome; experience; extracellular vesicles; genetic approach; graduate student; imaging platform; in vivo; insight; macromolecule; male; mass spectrometric imaging; meetings; microvesicles; mutant; novel; parent grant; plant fungi; prevent; professor; programs; rab GTP-Binding Proteins; research and development; skills; student mentoring; success; tumor progression; undergraduate student; vesicular release

Project summary This diversity supplement to R01 GM135433 proposes to train graduate student Malek Elsayyid as she uses the strengths of the C. elegans system to define factors that regulate extracellular vesicle (EV) biogenesis as well as the physiological significance of EV shedding. Secreted EVs, are membrane wrapped structures that share protein, RNA, lipid, and metabolite cargo with recipient cells. Released from most if not all cell types, EVs play a critical role during physiological processes and in pathological conditions. In C. elegans, EVs bud from sensory neuron cilia, then are taken up by surrounding glia or discharged into the environment. We developed transgenic worms that express different fluorescently tagged EV cargoes at single copy and optimized imaging parameters to visualize secreted EVs. The scientific goals of this supplement are to define the role of small G-protein RAB-28 in EV biogenesis and determine the impact of phagocytosed EVs on glia. This supplement proposal includes a mentoring plan to maximize Ms. Elsayyid’s training and future success. Performing research as an undergrad at a small liberal arts school limited her access to specialized equipment and technical expertise. Further, the COVID-19 pandemic has significantly impacted Ms. Elsayyid’s graduate experiences. In her training, Ms. Elsayyid will focus on four key areas. First, she will learn advanced super resolution and TIRF microscopy skills as she brings her project from start to completion. Second, she will use these data for scientific writing, honing communication skills by writing abstracts, research manuscripts, grant proposals, and a thesis dissertation. Third, she will present her research at C. elegans community, Chemistry Biology Interface program, DE Neuroscience, and national/international meetings. Finally, she will gain leadership skills mentoring students in the lab, a course-based research experience, and the community. Ms. Elsayyid will be advised by a mentoring team at the University of Delaware (UD). Dr. Jessica Tanis, Ms. Elsayyid’s primary mentor is an Assistant Professor in the Department of Biological Sciences with > 16 years C. elegans experience and 5 years studying EVs. Dr. Jeffrey Caplan, the Director of BioImaging at UD who studies transfer of EVs between plants and fungi has technical expertise in the imaging platforms and image analysis software programs required to train Ms. Elsayyid for this project. Dr. Grimes, a Professor in the Department of Chemistry and Biochemistry and co-director of the NIH-funded Chemistry Biology Interface (CBI) Program at UD will focus on Ms. Elsayyid’s career development, meeting with her bi-annually to review her IDP and informally on a weekly basis at CBI meetings. In conclusion, development of research expertise, communication skills, and mentoring experience as well as participation in the UD CBI program and professional networking opportunities will help Ms. Elsayyid achieve her career goal of becoming a professor, running a research lab and teaching in the classroom.

项目概要 R01 GM135433 的多样性补充建议培训研究生 Malek Elsayyid，因为她使用 线虫系统的优势将调节细胞外囊泡 (EV) 生物发生的因素定义为 以及 EV 脱落的生理意义 分泌的 EV 是膜包裹的结构。 与受体细胞共享大多数（如果不是全部）细胞类型释放的蛋白质、RNA、脂质和代谢物。 EV 在生理过程和病理条件下发挥着关键作用。 在线虫中，EV 萌芽。 来自感觉神经元纤毛，然后被周围的神经胶质细胞吸收或排放到环境中。 开发了转基因蠕虫，以单拷贝表达不同荧光标记的 EV 货物，并且 优化的成像参数以可视化分泌的 EV。本补充品的科学目标是定义。 小 G 蛋白 RAB-28 在 EV 生物合成中的作用，并确定吞噬 EV 对神经胶质细胞的影响。 该补充提案包括一项指导计划，旨在最大限度地提高 Elsayyid 女士的培训和未来的成功。 作为一名本科生在一所小型文理学院进行研究限制了她获得专业设备的机会 此外，COVID-19 大流行对 Elsayyid 女士的毕业生产生了重大影响。 在她的培训中，Elsayyid女士将重点关注四个关键领域：首先，她将学习高级超级。 其次，她将使用她的项目从开始到完成的分辨率和 TIRF 显微镜技能。 这些数据用于科学写作、通过撰写摘要、研究手稿、资助来磨练沟通技巧 第三，她将在秀丽隐杆线虫社区化学领域展示她的研究成果。 最后，她将获得生物界面项目、DE 神经科学和国内/国际会议。 在实验室、基于课程的研究经验和社区中指导学生的领导技能。 Elsayyid 女士将得到特拉华大学 (UD) 杰西卡·塔尼斯 (Jessica Tanis) 博士和 Elsayyid 的主要导师是生物科学系的助理教授，拥有超过 16 年的经验 UD 生物成像主任 Jeffrey Caplan 博士拥有线虫经验和 5 年研究经验。 研究 EV 在植物和真菌之间的转移 拥有成像平台和图像方面的技术专业知识 为该项目培训 Elsayyid 女士所需的分析软件程序。 化学与生物化学系和 NIH 资助的化学生物学接口联合主任 (CBI) UD 的项目将重点关注 Elsayyid 女士的职业发展，每年两次与她会面以进行审查 她的 IDP 以及每周一次的 CBI 会议上的非正式会议 总之，发展研究专业知识， 沟通技巧、指导经验以及参与 UD CBI 计划和 专业的人脉机会将帮助 Elsayyid 女士实现成为教授的职业目标， 运营研究实验室并在课堂上教学。