Human Specific Signaling Circuitry in Cone Precursor Development

锥体前体发育中的人类特异性信号传导电路

• 批准号: 9238776
• 负责人: David Cobrinik
• 金额: $ 41.63万
• 依托单位: CHILDREN'S HOSPITAL OF LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9238776
• 关键词: Animal Model; Animals; Biological Models; Blindness; Cell Differentiation process; Cell Lineage; Cells; Chromatin; Cone; Development; Disease; Disease model; Elements; Environment; Genes; Human; In Vitro; Individual; Knowledge; Lead; Light; Mediating; Mediator of activation protein; Mitotic; Modeling; Morphogenesis; Mus; Pathogenesis; Pathway interactions; Phototransduction; Portraits; Primates; Proteins; Recurrence; Regulatory Element; Research; Retina; Retinal; Retinal Cone; Retinal Diseases; Signal Pathway; Signal Transduction; Stem cells; Structure; System; Tissues; Vertebrates; Vision; cell type; differential expression; fetal; fovea centralis; human embryonic stem cell; improved; in vivo; insight; macula; novel strategies; novel therapeutics; precursor cell; programs; public health relevance; simulation; synaptogenesis; transcription factor; transcriptome; transcriptome sequencing

 DESCRIPTION (provided by applicant): Retinal diseases can lead to a devastating loss of vision and have grave personal and societal consequences. They are often modeled in animals, and more recently in human embryonic stem cell (hESC)-derived retina, in efforts to improve understanding of disease pathogenesis and to develop novel therapies. However, human retinal development differs from that of animal and hESC-derived retina models in important yet poorly understood ways. Defining such differences could reveal previously unrecognized human-specific features, identify developmentally important factors in the retina's in vivo ocular environment, and enable more accurate modeling of retinal diseases both in animals and in the hESC-derived retina system. This study aims to improve understanding of human retina features that fail to be modeled in mice or in hESC- derived retina, while also defining the differentiation states of a human retinal cell type in unprecedented detail. The study focuses on the post-mitotic differentiation of human cone photoreceptor precursors. These cells may be especially poorly represented in model systems because they form specialized structures (the macula and fovea) and express a proliferation-related program that is not evident in mouse or in hESC-derived models. We propose to define the differentiation states through which post-mitotic cone precursor's progress in vivo, in developing human and mouse retina, and also in vitro, in hESC-derived retina, and to explore the basis of major discrepant features of the model systems. In each context, we will a) isolate and define transcriptomes of individual cone precursors over a range of differentiation states, b) define and temporally order cone precursor differentiation states, and c) identify dynamically regulated genes and signaling pathways that mark the progression from one state to another. We will then probe the basis of human cone precursor cell state transitions by defining open chromatin regions and conserved cis-regulatory elements in dynamically regulated genes. We will also define the orthologous differentiation states through which human, mouse, and hESC-derived cone precursors progress and the major differences between the three settings, including but not limited to the human cone precursor proliferation-related program. Together, the studies will provide a detailed portrait of human cone precursor differentiation, enable development of more accurate in vivo and in vitro retinal disease models, and provide a novel approach to the study of human retinal development and disease.

 描述（由申请人提供）：视网膜疾病可导致毁灭性的视力丧失，并产生严重的个人和社会后果。它们通常以动物为模型，最近又以人类胚胎干细胞 (hESC) 来源的视网膜为模型，以努力增进对疾病发病机制的了解并开发新疗法。然而，人类视网膜发育与动物和 hESC 衍生的视网膜模型存在一些重要但人们知之甚少的不同之处。定义这些差异可以揭示以前未被识别的人类特异性特征，识别视网膜体内眼环境中发育重要的因素，并能够对动物和 hESC 衍生的视网膜系统中的视网膜疾病进行更准确的建模。这项研究旨在提高对无法在小鼠或 hESC 衍生视网膜中建模的人类视网膜特征的理解，同时以前所未有的细节定义人类视网膜细胞类型的分化状态。该研究的重点是人类视锥光感受器前体的有丝分裂后分化。这些细胞在模型系统中的表现可能特别差，因为它们形成特殊的结构（黄斑和中央凹）并表达与增殖相关的程序，而这在小鼠或 hESC 衍生模型中并不明显。我们建议定义有丝分裂后锥体前体在体内（在人类和小鼠视网膜发育过程中）以及在体外（在hESC衍生的视网膜中）进展的分化状态，并探索模型系统主要差异特征的基础。在每种情况下，我们将a）分离和定义一系列分化状态下各个视锥前体的转录组，b）定义并按时间排序视锥前体分化状态，以及c）识别标记从一种状态到另一种状态进展的动态调节基因和信号通路。然后，我们将通过定义开放染色质区域和动态调控基因中保守的顺式调控元件来探讨人类视锥细胞前体细胞状态转变的基础。我们还将定义人类、小鼠和 hESC 衍生的视锥细胞前体进展的直系同源分化状态，以及三种设置之间的主要差异，包括但不限于人类视锥细胞前体增殖相关程序。总之，这些研究将提供人类视锥细胞前体分化的详细描述，能够开发更准确的体内和体外视网膜疾病模型，并为人类视网膜发育和疾病的研究提供一种新方法。