Molecular mechanisms underlying spatial patterning of mammalian skin

项目摘要/摘要 协调早期胚胎发育的精确的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是 在基因水平上决定的，连接基因和表型的分子机制仍然存在 人们对此知之甚少。阐明这些机制对人类健康至关重要，因为 这些过程往往会导致各种发育缺陷和疾病状态。在这份提案中，我将研究 周期性色素图案(如条纹和斑点)的形成，以揭示其机制 在胚胎发育过程中，位置信息在哺乳动物的皮肤中被编码。周期性的色素图案出现 来自被编程为空间受限的非随机发育过程，因此 为了解组织如何获取位置信息提供了一个极好的模型。具体来说，我会 利用非洲条纹小鼠(横纹鼠)中自然进化的色素图案 Pumilio)，一种新兴的哺乳动物模型系统。我将结合各种基因组，转录，和 用功能方法剖析位置信息的分子和发育机制 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 将使用新的染色质图谱策略与功能方法相结合来识别顺式调控 控制色素沉着基因空间受限表达的区域及其相关因素。 其次，我将使用单细胞RNA测序来重建不同细胞的发育轨迹 胚胎皮肤的类型和识别早期建立模式边界的信号(S) 胚胎发生。然后，我将调节候选基因的表达，从功能上测试它们对色素的影响 图案的形成。综上所述，这项工作将大大促进我们对这些机制的理解 在哺乳动物早期发育过程中建立和实施空间模式。通过 结合使用新的哺乳动物模型系统和创新的尖端方法，这些 将对机制进行详细的研究，这在当前文献中是不存在的。这项工作将产生 对哺乳动物皮肤发育的基本过程的洞察，将为 了解发育障碍的机制基础，并产生更全面的 调控差异基因表达的机制概述，这是各种 人类的疾病和功能障碍。此外，我的顾问和合作顾问的专业知识结合在一起， 再加上普林斯顿大学丰富的资源(例如，流式细胞仪核心和高级 基因组学核心)，将允许我大大扩展我的科学训练；通过学习新的技能集(例如， 先进的基因组/转录组技术和生物信息学)到职业发展。

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.