Identifying mechanisms of coordinated outgrowth during zebrafish fin regeneration

确定斑马鱼鳍再生过程中协调生长的机制

• 批准号: 10315962
• 负责人: Victor Maxwell Lewis
• 金额: $ 6.64万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10315962
• 关键词: Acute; Biological Assay; Cell Nucleus; Cell Proliferation; Cells; Chromatin; Communication; Communities; Complex; Cues; Data Set; Defect; Development; Developmental Biology; Distal; Environment; Epidermis; Epithelial; Epithelial Attachment; Equipment; Excision; Fibroblast Growth Factor; Genes; Genetic; Genetic Transcription; Genomics; Glucocorticoids; Goals; Growth; Growth Factor; Human; Image; In Vitro; Injury; Institutes; Insulin; International; Intrinsic factor; Lead; Limb structure; Maps; Mentors; Mesenchymal; Mesenchyme; Modeling; Molecular; Molecular Biology; Natural regeneration; Oregon; Organ; Osteoblasts; Outcome; Pathway interactions; Phase; Population; Primary Cell Cultures; Principal Investigator; RNA; Receptor Signaling; Regenerative Medicine; Research; Research Training; Role; Signal Pathway; Signal Transduction; Source; Supervision; Swimming; Testing; Therapeutic; Time; Tissues; Training; Transposase; Universities; WNT Signaling Pathway; Work; Writing; Zebrafish; appendage; blastema; bone; career; cell behavior; cell type; chemical genetics; design; developmental genetics; empowered; healing; improved; in vivo; in vivo evaluation; injury and repair; innovation; insight; novel; organ injury; organ regeneration; osteoblast differentiation; regenerative; repaired; response; restoration; seal; single cell sequencing; single cell technology; single-cell RNA sequencing; skills; spatiotemporal; teacher; transcriptomics; wound epidermis

PROJECT SUMMARY This project will provide Dr. Victor Lewis with postdoctoral research training in vertebrate developmental genetics and genomics to prepare for a career as an academic principal investigator. The applicant will follow a multi-faceted research and training plan at the University of Oregon under supervision of lead sponsor Dr. Kryn Stankunas with additional mentoring from co-sponsor Dr. John Postlethwait. A fundamental goal of developmental biology is to identify the cellular and molecular mechanisms synchronizing cell behaviors across lineages for complex organ regeneration. Zebrafish fins perfectly recapitulate their original form following resection, making them a striking example of and tractable research model for appendage regeneration. An injury-induced de-differentiated but organized mass of cells called the blastema produces replacement fin cells and directs regenerative outgrowth. Fin outgrowth then depends on spatially segregated proliferation and differentiation activities that progressively restore organized tissue, including ray bones integrated with stump tissue. The major signaling pathways, the signal-producing and responding cells, and their general roles (e.g. growth vs. differentiation) are becoming appreciated. However, how signals enable spatiotemporally coordinated actions across cell types remains largely mysterious. Empowered by new single cell transcriptomics, Dr. Lewis will test a novel hypothesis that a regulated mesenchymal to epithelial-like cell state transition forms the outgrowth-promoting population of the distal blastema. He will also test a hypothesis how systemic factors coordinate osteoblast differentiation with fin outgrowth. Finally, Dr. Lewis will map the cell-type specific regulatory landscape of fin regeneration by integrating sophisticated single cell technologies with chemical genetics providing acute pathway inhibition. Collectively, this proposal will explore and characterize innovative models and mechanisms for coordinated outgrowth during appendage regeneration. Broadly, these insights will impact design of regenerative medicine approaches and provide a framework to understand appendage injury repair. Dr. Lewis’s training plan will broaden single-cell sequencing and technical research skills, 2) improve writing and communication skills, 3) develop skills as a mentor and teacher, and 4) engage in professional development activities to build skills to succeed as a principal investigator. The UO, Institute of Molecular Biology, and UO’s internationally known zebrafish research community provide facilities, equipment, and intellectual environment ideally aligned with Dr Lewis’s postdoctoral research and training goals.

项目总结 该项目将为维克多·刘易斯博士提供脊椎动物发育方面的博士后研究培训 遗传学和基因组学，为成为一名学术首席研究员做准备。申请者将遵循以下规则 俄勒冈大学在主要赞助商Kryn博士的监督下的多方面研究和培训计划 斯坦库纳斯，联合赞助人约翰·波斯尔斯韦特博士提供了额外的指导。的根本目标是 发育生物学是确定使细胞行为同步的细胞和分子机制。 复杂器官再生的谱系。斑马鱼的鳍完美地再现了它们最初的形状 切除，使它们成为附件再生的显著例子和易于处理的研究模型。一个 损伤诱导的去分化但有组织的细胞团，称为胚泡，产生替代的鳍细胞 并引导再生生长。然后，FIN的扩展依赖于空间隔离的扩散和 逐渐恢复有组织的组织的分化活动，包括与残肢整合的放射骨 组织。主要的信号通路、信号产生细胞和反应细胞及其一般作用(如： 增长与差异化)正变得越来越受重视。然而，信号如何在时空上启用 不同细胞类型之间的协调行动在很大程度上仍然是个谜。由新的单电池提供支持 刘易斯博士将测试一种新的假设，即受调控的间质到上皮样细胞状态 过渡期形成了远端胚泡促进生长的种群。他还将检验一项假设，即 全身因素协调成骨细胞分化和鳍的生长。最后，刘易斯博士将绘制细胞类型图 通过将复杂的单电池技术与 提供急性通路抑制的化学遗传学。总体而言，这项提案将探索和描述 在附肢再生过程中协调生长的创新模式和机制。大体上说，这些 洞察力将影响再生医学方法的设计，并提供一个框架来理解 附件损伤修复。刘易斯博士的培训计划将扩大单细胞测序和技术研究 技能，2)提高写作和沟通能力，3)发展作为导师和教师的技能，以及4)参与 专业发展活动，以培养作为首席调查员取得成功的技能。密歇根州理工学院 分子生物学和UO国际知名的斑马鱼研究社区提供设施、设备、 和智力环境与刘易斯博士的博士后研究和培训目标理想地保持一致。