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

项目摘要/摘要 视网膜发育发生在常见的视网膜前体细胞分化为七类 视网膜神经元和支持细胞组织成视网膜的三层。这一发展 这一过程主要是在Mace中研究的，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的详细机制的能力造成了很大的限制 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。视网膜疾病可导致功能障碍和死亡 特定的视网膜细胞类型，导致患者失明和失明。许多与视网膜相关的基因 疾病也与正常的视网膜发育有关，以及对正确分子的了解 过程将为在疾病状态下观察到的异常过程提供背景。最近，人类干细胞- 衍生的视网膜类有机化合物已成为研究细胞和分子过程的既定模型 视网膜发育的影响。这项研究的目标是同时进行RNA测序和谱系追踪 在体内小鼠视网膜和体外发育单细胞分辨率的人视网膜有机物质中 创建人类和小鼠视网膜形成的详细地图。拟议的研究将建立在我们的 了解小鼠作为视网膜研究的模式生物，并允许识别基因和 在人类视网膜发育过程中，细胞命运的精确决定所涉及的通路。终极的 对这两个模型系统的比较可能会进一步突出和区分关键重要基因的优先顺序。这 知识将有助于开发和改进治疗视网膜疾病的治疗策略。 该提案解释了一项研究战略和培训计划，以培养首席调查员克莱尔女士。 温格，在她整个研究生教育期间。温格女士已经完成了她研究生的第三年学业。 约翰霍普金斯大学医学院的人类遗传学博士前培训项目。毕业后，它是 她的目标是继续从事视觉研究，并开始一项研究的首席研究员职业生涯- 以制度为基础的。温格女士的教育必须是彻底的和跨学科的，才能发展她的技能 在分子和细胞生物学、生物信息学、负责任的研究、科学交流和 师徒关系。她的培训计划包括课程、研讨会、会议和实验室技能，由 各式各样的导师。拟议的研究和培训战略将为温格女士提供 在她的研究生生涯中脱颖而出并成功实现她未来目标的必要工具。

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.