Genetic and Cellular Mechanisms of Engrailed-1 Regulation and Function in Eccrine Sweat Gland Development

项目摘要 外分泌腺是人体皮肤的主要附属物，是人体体温调节所必需的。尽管他们 尽管它们具有重要意义，但对其发育的细胞和分子机制知之甚少。这项建议 建立在我们的工作涉及转录因子Engrailed-1作为一个独特的决定因素和驱动器的外分泌 腺形成，以确定诱导的规范和分化的遗传和细胞机制， 皮肤中的外泌汗腺。因此，本提案的目标是鉴定转录因子 在外分泌腺发育过程中直接调节Engrailed1表达（Aim1），并确定 下游Engrailed 1依赖性转录程序，然后执行外分泌腺形成（Aim2）。 该建议整合了体内测试和高通量发现的细胞和分子组成部分， 使用新产生的和现有的小鼠模型结合机械的外分泌腺发育 在培养的人皮肤细胞中的解剖以及新的小鼠和人单核之间的交叉验证 转录组数据集。因此，拟议的实验将描绘小汗腺发育 计划，通知努力建立一个体外系统，在其中研究外分泌腺，并提高 更广泛地了解外胚层附属物的命运和形成是如何在皮肤中建立的。从 从临床的角度来看，我们的发现将为未来的研究提供一个分子蓝图，以再生外分泌 腺体在体内，也提供了新的干预点，修复外分泌腺时，这些器官是 病理性改变或者损坏。

PROJECT SUMMARY Eccrine glands are the major appendage of human skin and required for human thermoregulation. Despite their importance, the cellular and molecular mechanisms of their development are poorly understood. This proposal builds on our work implicating the transcription factor Engrailed-1 as a unique determinant and driver of eccrine gland formation to define genetic and cellular mechanisms that induce the specification and differentiation of eccrine sweat glands in the skin. Accordingly, the goals of this proposal are to identify the transcription factors that directly modulate Engrailed1 expression during eccrine gland development (Aim1) and to define the downstream Engrailed 1-dependent transcriptional program that then executes eccrine gland formation (Aim2). This proposal integrates in vivo testing and high-throughput discovery of cellular and molecular components of eccrine gland development using both newly generated and existing mouse models combined with mechanistic dissection in cultured human skin cells and cross-validation between novel mouse and human single nucleus transcriptomic datasets. As such, the proposed experiments will delineate the eccrine gland developmental program, inform efforts to establish an in vitro system in which to study eccrine glands, and enhance understanding of how ectodermal appendage fate and formation are established in the skin more broadly. From a clinical perspective, our findings will provide a molecular blueprint for future studies to regenerate eccrine glands in vivo and also offer novel points of intervention for repair of eccrine glands when these organs are pathologically altered or damaged.