Molecular mechanisms underlying spatial patterning of mammalian skin

项目总结/摘要 协调早期胚胎发育的精确空间和时间模式机制 代表了发育生物学中的一个基本问题。虽然如此复杂的监管最终 在基因型水平上确定，连接基因型与表型的分子机制仍然存在， 不太了解。阐明这些机制对人类健康至关重要，因为 这些过程常常导致各种发育缺陷和疾病状态。在这份提案中，我将研究 周期性颜料图案的形成（例如，条纹和斑点）来揭示 位置信息在胚胎发生期间编码在哺乳动物皮肤中。周期性的色素图案出现 从非随机的发展过程，被编程为空间约束，从而 表示用于理解组织如何获取位置信息的极好模型。具体来说，我会 利用非洲条纹鼠（Rhabdomys）自然进化的色素模式， pumilio），一种新兴的哺乳动物模型系统。我将联合收割机结合各种基因组，转录组， 功能的方法来剖析分子和发育机制， 是在发育中的组织中获得和解释的，这是发育生物学中的一个长期问题。首先我 将使用新的染色质分析策略结合功能方法来识别顺式调节蛋白， 区域及其相关因素，控制色素沉着基因的空间限制表达。 其次，我将使用单细胞RNA测序来重建不同细胞的发育轨迹， 类型的胚胎皮肤，并确定信号，建立模式边界早在 胚胎发生然后，我将调节候选基因的表达，以功能测试它们对色素的影响 图案形成总之，这项工作将大大推进我们对机制的理解， 在哺乳动物早期发育过程中建立和实施空间模式。通过 结合使用一种新的哺乳动物模型系统和创新的，尖端的方法， 机制将被调查的详细程度不存在在目前的文献。这项工作将产生 深入了解哺乳动物皮肤发育的基本过程，将为以下方面提供一个框架： 了解发育障碍的机制基础，并产生更全面的 概述了调节差异基因表达的机制，这是各种研究的最前沿过程。 人类疾病和功能障碍。此外，我的顾问和共同顾问的专业知识， 再加上普林斯顿大学的丰富资源（例如，流式细胞仪核心和高级 基因组学核心），将使我大大扩展我的科学训练;从学习新的技能（例如， 先进的基因组/转录组技术和生物信息学）的职业发展。

Project Summary/Abstract The precise spatial and temporal patterning mechanisms that coordinate early embryonic development represent a fundamental question in developmental biology. While such complex regulation is ultimately determined at the genotypic level, the molecular mechanisms that connect genotype to phenotype remain poorly understood. Elucidation of these mechanisms is of critical importance to human health, as errors in these processes often lead to various developmental defects and diseased states. In this proposal, I will study the formation of periodic pigment patterns (e.g., stripes and spots) to uncover the mechanisms by which positional information is encoded in mammalian skin during embryogenesis. Periodic pigment patterns arise from nonrandom developmental processes that are programmed to be spatially constrained, and thereby represent an excellent model for understanding how a tissue acquires positional information. Specifically, I will take advantage of the naturally evolved pigment pattern seen in the African striped mouse (Rhabdomys pumilio), an emerging mammalian model system. I will combine a variety of genomic, transcriptomic, and functional approaches to dissect the molecular and developmental mechanisms by which positional information is acquired and interpreted in developing tissues, a long-standing question in developmental biology. First, I will use novel chromatin profiling strategies coupled to functional approaches to identify the cis-regulatory regions and their associated factors that control the spatially restricted expression of pigmentation genes. Second, I will use single-cell RNA sequencing to reconstruct the developmental trajectories of the different cell types of the embryonic skin and identify the signal(s) that establish the pattern boundary early in embryogenesis. I will then modulate candidate gene expression to functionally test their effects on pigment pattern formation. Taken together, this work will substantially advance our understanding of the mechanisms that both establish and implement spatial patterns during early mammalian development. Through the combined use of a novel mammalian model system and innovative, cutting-edge approaches, these mechanisms will be investigated in a level of detail non-existent in the current literature. This work will generate insights into the basic processes governing the development of mammalian skin, will provide a framework for understanding the mechanistic basis of developmental disorders, and generate a more comprehensive overview of the mechanisms regulating differential gene expression, a process at the forefront of various human diseases and dysfunctions. Furthermore, the combined expertise of my advisor and co-advisor, coupled with the abundant resources available at Princeton University (e.g., flow cytometry core and advanced genomics core), will allow me to substantially expand my scientific training; from learning new skillsets (e.g., advanced genomic/transcriptomic techniques and bioinformatics) to career development.