In toto imaging and genomics to decode ear hair cell formation and regeneration

描述(申请人提供)：听力和平衡的丧失在人类中是一种普遍的和衰弱的医学状况，主要是由毛细胞的丧失引起的，毛细胞是内耳的主要感觉细胞。人类的毛细胞不会自然再生，这与斑马鱼等其他动物不同，斑马鱼的毛细胞很容易再生。为了了解毛细胞在发育过程中的形成和毛细胞在损伤后的再生，我们将采用基于系统生物学的综合方法。我们的方法集成在TOTO成像中，它提供了跨越毛细胞生成的空间和时间的系统高分辨率分析，以及允许对整个基因组的转录活动进行系统分析的基因组方法。具体地说，我们将使用TOTO成像技术，这是我们开发的一项技术，以生成一个基于细胞的4维数字耳朵，它全面量化斑马鱼毛细胞的形成和再生的细胞过程。我们将使用特定细胞类型的组蛋白修饰芯片序列来确定在毛细胞生成的所有关键步骤中，活跃在整个基因组中的增强剂、启动子和绝缘体。生物信息学方法将被用来将定义的增强子内的转录因子结合位点映射到它们控制的基因，以在我们的数字耳朵的虚拟细胞内构建一个全面的顺式调控网络。这项研究将对基因组如何编码毛细胞的逐步规范提供前所未有的见解，对人类毛细胞再生具有潜在的重要意义。与健康的相关性听力和平衡方面的缺陷是普遍存在的，并使人虚弱。它们主要是由于内耳毛发的感觉细胞(毛细胞)的丧失造成的，内耳毛发不能在人类身上再生，但在其他动物中可以再生。我们试图了解内耳毛细胞再生的遗传和细胞控制。 与公共卫生相关：听力和平衡方面的缺陷普遍存在，并使人虚弱。它们主要是由于内耳毛发的感觉细胞(毛细胞)的丧失造成的，内耳毛发不能在人类身上再生，但在其他动物中可以再生。我们试图了解内耳毛细胞再生的遗传和细胞控制。

DESCRIPTION (provided by applicant): Loss of hearing and balance is a widespread and debilitating medical condition in humans, and is predominantly caused by a loss of hair cells, the primary sensory cell of the inner ear. Hair cells do not naturally regenerate in humans, in contrast to other animals such as zebrafish where hair cells readily regenerate. To understand hair cell formation during development and hair cell regeneration in response to damage we will undertake an integrative systems biology based approach. Our approach integrates in toto imaging which provides systematic high resolution analysis across the space and time of hair cell generation with omic approaches that allow systematic analysis of transcriptional activity across the genome. Specifically, we will use in toto imaging, a technology we developed, to generate a 4-dimensional, cell-based Digital Ear that comprehensively quantifies the cellular processes that form and regenerate hair cells in zebrafish. We will use cell-type-specific ChIP-seq of histone modifications to determine the enhancers, promoters, and insulators active across the entire genome at all the key steps of hair cell generation. Bioinformatic approaches will be used to map transcription factor binding sites within defined enhancers to the genes they control to construct a comprehensive cis-regulatory network within the virtual cells of our Digital Ear. This research will provide unprecedented insight into how the genome encodes the stepwise specification of hair cells, with potentially important implications for hair cell regeneration in humans. Relevance to Health Deficiencies in hearing and balance are widespread and debilitating. They are principally caused by loss of the sensory cells (hair cells) of the inner ear hair which cannot regenerate in humans but can in other animals. We seek to understand the genetic and cellular control of inner ear hair cell regeneration. PUBLIC HEALTH RELEVANCE: Deficiencies in hearing and balance are widespread and debilitating. They are principally caused by loss of the sensory cells (hair cells) of the inner ear hair which cannot regenerate in humans but can in other animals. We seek to understand the genetic and cellular control of inner ear hair cell regeneration.