Cranial placode induction from human embryonic stem cells

描述(申请人提供)：颅骨胎盘是短暂的胚胎结构，最终发育成感觉器官和主要内分泌腺--脑下垂体前叶的一部分。这些安慰剂来自外胚层，外胚层是三个胚层之一，众所周知，外胚层可以形成表皮、大脑和脊髓。安慰剂在发育中的大脑和表皮两侧的一个小区域内发育。目前，我们对这些瞬时结构是如何在人体内发展的了解甚少。为了更好地了解这一过程，该项目旨在确定将外胚层限制为非神经命运的潜在分子机制，并允许它发育成不同的安慰剂。由于这些组织产生的早期时间点，将使用人类胚胎干细胞来模拟发育。干细胞将在不同的培养条件下确定最佳条件，首先是非神经性外胚层，然后是胎盘前外胚层，最后是不同的安慰剂。所有这些组织都由一组不同的基因定义，这些基因将被分析以在单细胞分辨率下共同表达。这些知识可以用来研究基因缺陷是如何改变胎盘发育的。作为这方面的一步，我们将设计一个EYA1基因敲除的人类胚胎干细胞系，并在单细胞分辨率下分析这些细胞在胎盘发育过程中转录谱的变化。我们从这项研究中学到的对于研究影响感官系统的人类疾病具有广泛的意义，包括内耳。

DESCRIPTION (provided by applicant): The cranial placodes are transient embryonic structures that eventually develop into portions of the sense organs and the anterior pituitary, a major endocrine gland. These placodes arise from the ectoderm, one of the three germ layers, which is known to give rise to the epidermis, brain, and spinal cord. The placodes develop in a small region that is flanked by the developing brain and epidermis. Currently we understand very little of how these transient structures develop in humans. To gain better insight into this process, this project aims to determine the underlying molecular mechanisms that restrict the ectoderm to a non-neural fate and allow it to develop into the different placodes. Due to the early time points in which these tissues arise, development will be modeled using human embryonic stem cells. Stem cells will be subjected to different culture conditions to determine the optimal conditions to derive first non-neural ectoderm, followed by pre-placodal ectoderm and finally the different placodes. All of these tissues are defined by a distinct set of genes, which will be analyzed for co-expression at single cell resolution. This knowledge can be used to study how defects in genes alter placode development. As a step towards this, we will engineer an EYA1 knockout human embryonic stem cell line and assay changes in the transcriptional profile of these cells during placode development at single cell resolution. What we learn from this research has broad implications for study of human disease affecting the sensory organ systems, including the inner ear.