Investigation of a neuromesendodermal progenitor population in the posterior avian endoderm

禽类后内胚层神经中内胚层祖细胞群的研究

基本信息

批准号：
10456910
负责人：
Nandan L Nerurkar
金额：
$ 39.96万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10456910
关键词：
Adult Atlases Biochemical Biophysics Birds Brain Cell Line Cells Chick Embryo Complex Cues Development Developmental Biology Dorsal Ectoderm Ectoderm Cell Embryo Embryology Embryonic Development Endoderm Endoderm Cell Genes Germ Layers Goals Invaded Investigation Malignant Neoplasms Mesenchyme Mesoderm Mesoderm Cell Multipotent Stem Cells Neural tube Organ Pathogenesis Pattern Population Process Regulation Research Shapes Signal Transduction Source Specific qualifier value Stereotyping Surface Time Tissues Totipotent Tube Work cell behavior directed differentiation epithelial to mesenchymal transition experimental study gastrulation in vivo Model notochord novel pluripotency progenitor programs prospective segregation single-cell RNA sequencing stem stem cell population stem cells transcriptomics

Adult Atlases Biochemical Biophysics Birds Brain Cell Line Cells Chick Embryo Complex Cues Development Developmental Biology Dorsal Ectoderm Ectoderm Cell Embryo Embryology Embryonic Development Endoderm Endoderm Cell Genes Germ Layers Goals Invaded Investigation Malignant Neoplasms Mesenchyme Mesoderm Mesoderm Cell Multipotent Stem Cells Neural tube Organ Pathogenesis Pattern Population Process Regulation Research Shapes Signal Transduction Source Specific qualifier value Stereotyping Surface Time Tissues Totipotent Tube Work cell behavior directed differentiation epithelial to mesenchymal transition experimental study gastrulation in vivo Model notochord novel pluripotency progenitor programs prospective segregation single-cell RNA sequencing stem stem cell population stem cells transcriptomics

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项目摘要

PROJECT SUMMARY/ABSTRACT Transformation of the early embryo from a seemingly disorganized ball of stem cells to the complex and precisely patterned adult form requires stereotyped regulation cell behaviors via biophysical and biochemical cues. Using the chick embryo, the long term goals of my research program are to understand from a highly quantitative perspective how this is orchestrated. Approximately half of the lab focuses on mechanobiology of embryonic development, seeking to understand the forces that shape tissues and organs in the embryo and the cues that specify those forces. The remainder of the lab studies cell fate determination, either in the context of early brain development or in the developing gut. The present application stems from the latter. During embryonic development, one of the first major steps in lineage segregation is gastrulation, when a single totipotent layer of cells separates into three germ layers, endoderm, mesoderm, and ectoderm. While conventionally, it has long been accepted that all progenitors of each germ layer are generated only during gastrulation, recent studies have challenged this, identifying a multipotent progenitor population that persists after gastrulation ends in the tailbud. Within this population, known as neuromesodermal progenitors, a single cell can give rise to progeny that contribute to both notochord (mesoderm) and neural tube (ectoderm). While studying gut tube formation in the chick embryo, we recently identified a population of endoderm cells that line the ventral surface of the tailbud, and appear to undergo a widespread and previously unreported epithelial-to- mesenchymal transition (EMT). These cells invade the neighboring tailbud tissue, reaching as far as the dorsal surface of the tailbud. Importantly, cells can be seen dispersed throughout the region where neuromesodermal progenitors are known to reside, as well as in the neural tube and notochord. This suggests that these endodermal cells represent an unappreciated source of neuromesodermal progenitors, which, if they retain the ability to generate endodermal tissues as well, would be remarkable. The present application combines classical embryology approaches such as fate mapping and grafting experiments with gene misexpression and single-cell RNA Sequencing in an effort to make sense of the simple yet puzzling observation that endoderm cells undergo a localized EMT and invade the tailbud at a time when gastrulation is complete. The proposal aims to identify the tissues to which this prospective progenitor population contributes, and to construct a detailed single-cell transcriptomic atlas of this population, from which identity, regulatory circuits, and proposed lineage trajectories can be inferred. At the completion of the proposed work, we will have established a new in vivo model to study endodermal EMT, and potentially identified a novel stem cell population residing in the endoderm, with the ability to give rise to mesodermal and ectodermal cells. This stands in contrast to long-held notions of lineage segregation during embryonic development, and therefore has the potential to be transformative for the fields of developmental biology and stem cell reprogramming.

项目摘要/摘要早期胚胎从看似混乱的干细胞球转变为复合物和精确的成年形式需要通过生物物理和生化的刻板印象调节细胞行为提示。使用小鸡胚胎，我的研究计划的长期目标是从高度了解定量观点如何精心策划。大约一半的实验室专注于机械生物学胚胎发育，寻求了解胚胎中组织和器官的力量指定这些力量的提示。其余的实验室研究细胞命运确定，要么在上下文中早期大脑发育或发展中的肠道。目前的应用源于后者。在胚胎发育期间，谱系隔离的第一个主要步骤之一是胃肠道，当单个细胞的单位层分成三个细菌层，内胚层，中胚层和外胚层。尽管通常，长期以来，人们一直认为，每个细菌的所有祖细胞仅在胃分解，最近的研究对此提出了质疑，确定了持续存在的多元祖细胞种群胃结束后，尾巴以尾巴结束。在该人群中，称为神经溶性祖细胞，一个细胞会引起后代，从而有助于脊索（中胚层）和神经管（外胚层）。尽管在研究雏鸡胚胎中的肠道形成时，我们最近确定了内胚层细胞的群体尾梁的腹表面，似乎经历了广泛的，以前未报告的上皮到 - 间充质转变（EMT）。这些细胞侵入相邻的尾bud组织，到达背侧尾舱的表面。重要的是，可以看到细胞分散在整个区域，神经抑制作用已知祖细胞居住，以及在神经管和脊索中。这表明这些内皮细胞代表神经抑制术的未欣赏来源，如果它们保留了产生内皮组织的能力也将是显着的。当前的应用程序结合了经典的胚胎学方法，例如命运图和嫁接实验，并具有基因的作用和单细胞RNA测序以理解内胚层的简单而令人困惑的观察细胞经过局部EMT，并在胃胃完成时侵入尾b。提案旨在确定该前瞻性祖细胞贡献的组织，并构建该人群的详细的单细胞转录组图集，从中身份，调节回路和提议可以推断谱系轨迹。拟议的工作完成后，我们将建立一个新的研究内皮EMT的体内模型，并有可能确定存在于内胚层具有产生中胚层和外胚层细胞的能力。这与长期存在相反胚胎发育过程中谱系隔离的概念，因此有潜力发育生物学和干细胞重编程领域的变革性。