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.

项目摘要 R 01 GM 135433的多样性补充建议培训研究生Malek Elsayyid，因为她使用 C. elegans系统来定义调节细胞外囊泡（EV）生物发生的因子， 以及EV脱落的生理意义。分泌的EV是膜包裹的结构， 与受体细胞共享蛋白质、RNA、脂质和代谢物货物。从大多数细胞中释放出来， EV在生理过程和病理条件下发挥关键作用。In C. elegans，EV芽 然后被周围的神经胶质细胞吸收或释放到环境中。我们 开发了以单拷贝表达不同荧光标记EV货物的转基因蠕虫， 优化成像参数以可视化分泌的EV。本补充的科学目标是定义 小G蛋白RAB-28在EV生物发生中的作用，并确定吞噬的EV对神经胶质的影响。 这一补充建议包括一项指导计划，以最大限度地提高Elsayyid女士的培训和未来的成功。 在一所小型文科学校读本科时进行研究限制了她获得专业设备的机会 和技术专长。此外，COVID-19大流行严重影响了Elsayyid女士的毕业生 经验在培训中，Elsayyid女士将侧重于四个关键领域。首先，她将学习高级超 分辨率和TIRF显微镜技能，因为她把她的项目从开始到完成。第二，她将利用 这些数据用于科学写作，通过撰写摘要、研究手稿、资助 提案和论文。第三，她将在C。线虫群落，化学 生物学接口程序，DE神经科学，和国家/国际会议。最后，她将获得 领导技能指导学生在实验室，基于课程的研究经验，和社区。 女士Elsayyid将由特拉华州大学（UD）的一个指导小组提供建议。杰西卡·塔尼斯博士， 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。 C. elegans经验和5年研究电动汽车。Jeffrey Caplan博士，UD生物成像总监， 研究植物和真菌之间的EV转移在成像平台和图像方面具有技术专长 培训Elsayyid女士进行该项目所需的分析软件程序。格莱姆斯博士， 化学和生物化学系，NIH资助的化学生物学接口的共同主任 (CBI)UD的计划将侧重于Elsayyid女士的职业发展，每两年与她会面一次， 她的国内流离失所者和非正式每周在加州调查局会议。总之，发展研究专门知识， 沟通技巧，指导经验以及参与UD CBI计划， 职业社交机会将帮助Elsayyid实现成为教授的职业目标， 管理一个研究实验室，在课堂上教学。