Simultaneous Single-cell Lineage Tracing and Transcriptome Analysis for Mapping Human and Mouse Retinal Development

同时进行单细胞谱系追踪和转录组分析，绘制人类和小鼠视网膜发育图谱

• 批准号: 10161599
• 负责人: Claire Bell
• 金额: $ 4.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10161599
• 关键词: Animal Model; Bioinformatics; Biological; Biological Models; Blindness; Cell Differentiation process; Cell Lineage; Cell Therapy; Cells; Cessation of life; Communication; Complex; Data; Data Set; Development; Developmental Process; Disease; Education; Fetal Tissues; Functional disorder; Future; Gene Expression; Gene Targeting; Genes; Genetic; Goals; Graduate Education; Human; Human Genetics; Impairment; In Vitro; Institution; Knowledge; Laboratories; Lead; Maps; Mentors; Mentorship; Methods; Modeling; Molecular; Molecular Profiling; Molecular and Cellular Biology; Mus; Myristica fragrans; Neurons; Nuclear; Organoids; Pathway interactions; Patients; Pharmaceutical Preparations; Principal Investigator; Process; Rattus; Research; Research Training; Resolution; Retina; Retinal Diseases; Stem Cell Development; Study models; Supporting Cell; System; Technology; Therapeutic; Tissue-Specific Gene Expression; Training; Training Programs; Vision research; base; career; cell type; genome editing; genomic locus; graduate student; human fetus tissue; human stem cells; improved; in utero; in vivo; insight; medical schools; meetings; molecular drug target; novel; novel strategies; pre-doctoral; prevent; responsible research conduct; retinal neuron; retinal progenitor cell; single cell analysis; skills; spine bone structure; stem cell biology; stem cells; targeted treatment; tool; transcriptome; transcriptome sequencing; transcriptomics; transdifferentiation

Project Summary/Abstract Retinal development occurs as a common retinal progenitor cell is differentiated into the seven classes of retinal neurons and supporting cells that organize into the three layers of the retina. This developmental process has primarily been studied in mace, as it takes place largely in utero, and the difficulty of obtaining human fetal tissue has put significant limitations on the ability to study the detailed mechanisms of early human retinal development. Understanding the genetic complexity of retinal cell fate decisions and cell lineages holds implications for our understanding of retinal disease. Retinal diseases can cause the dysfunction and death of specific retinal cell types, leading to vision loss and blindness in patients. Many genes associated with retinal diseases have also been implicated in normal retinal development, and knowledge of the correct molecular processes will give context to abnormal processes observed in disease states. Recently, human stem cell- derived retinal organoids have become an established model for studying the cellular and molecular processes of retinal development. The goal of this study is to perform simultaneous RNA sequencing and lineage tracing in vivo in murine retinas and in vitro in developing human retinal organoids at single-cell resolution in order to create detailed maps of human and mouse retina formation. The proposed study will build upon our understanding of mice as a model organism for retinal studies and allow the identification of genes and pathways involved in the precise cell fate decisions that occur during human retinal development. The ultimate comparison of these two model systems may further highlight and prioritize genes of key importance. This knowledge will help to develop and improve therapeutic strategies for treating retinal diseases. This proposal explains a research strategy and training plan to develop the principal investigator, Ms. Claire Wenger, throughout her graduate education. Ms. Wenger has completed her third-year as a graduate student in the Human Genetics Predoctoral Training Program at Johns Hopkins School of Medicine. After graduating, it is her goal to continue to pursue vision research and embark on a career as a principal investigator at a research- based institution. Ms. Wenger’s education must be thorough and interdisciplinary in order to develop her skills in molecular and cellular biology, bioinformatics, responsible conduct of research, scientific communication, and mentorship. Her training plan includes coursework, seminars, meetings, and laboratory skills taught to her by a diverse set of mentors. The proposed research and training strategies will provide Ms. Wenger with the necessary tools to excel in her graduate career and successfully attain her future goals.

项目总结/摘要 视网膜发育发生在普通视网膜祖细胞分化成七类视网膜细胞的过程中。 视网膜神经元和支持细胞组成视网膜的三层。这种发育 这个过程主要是在梅斯研究，因为它主要发生在子宫内，而且很难获得 人类胎儿组织对研究早期人类胚胎发育的详细机制的能力有很大的限制。 视网膜发育了解视网膜细胞命运决定和细胞谱系的遗传复杂性 对我们理解视网膜疾病的影响。视网膜疾病可导致视网膜功能障碍和死亡。 特定的视网膜细胞类型，导致患者视力丧失和失明。许多与视网膜病变相关的基因 疾病也与正常的视网膜发育有关， 过程将为疾病状态中观察到的异常过程提供背景。最近，人类干细胞- 衍生的视网膜类器官已经成为研究细胞和分子过程的既定模型 视网膜的发育。本研究的目的是同时进行RNA测序和谱系追踪 在鼠视网膜中的体内和在体外以单细胞分辨率发育的人视网膜类器官中， 绘制人类和老鼠视网膜形成的详细地图。拟议的研究将建立在我们的 理解小鼠作为视网膜研究的模式生物，并允许鉴定基因和 这是人类视网膜发育过程中参与细胞命运决定的关键途径。最终 这两个模型系统的比较可以进一步突出和优先化关键重要性的基因。这 知识将有助于发展和改进治疗视网膜疾病的治疗策略。 这份提案解释了一项研究战略和培训计划，以培养首席研究员克莱尔女士。 温格，在她的研究生教育。温格女士已经完成了她的第三年作为一个研究生， 约翰霍普金斯医学院的人类遗传学博士前培训项目。毕业后， 她的目标是继续追求视觉研究，并开始作为一个研究的首席研究员的职业生涯- 基于机构。温格女士的教育必须是彻底的和跨学科的，以发展她的技能 在分子和细胞生物学，生物信息学，负责任的研究行为，科学交流， 导师制她的培训计划包括课程作业、研讨会、会议和由一位 不同的导师。拟议的研究和培训战略将为温格女士提供 必要的工具，在她的研究生生涯中脱颖而出，并成功地实现她的未来目标。