Molecular and Cell Biology of Neuromesodermal Progenitors

神经中胚层祖细胞的分子和细胞生物学

• 批准号: 10689582
• 负责人: Benjamin L Martin
• 金额: $ 39.03万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10689582
• 关键词: Behavior; Biological; Biology; Biosensor; Cell Cycle; Cell Differentiation process; Cell Maintenance; Cell model; Cells; Cellular biology; Cyclin-Dependent Kinases; Development; Disease; Ectoderm; Embryo; Family; G1 Phase; G2 Phase; Gene Expression; Generations; Germ Layers; In Vitro; Invaded; Knowledge; Maintenance; Malignant Neoplasms; Mediating; Mesoderm; Mesoderm Cell; Molecular; Molecular Biology; Normal Cell; Pathway interactions; Phase; Play; Postdoctoral Fellow; Receptor Activation; Regenerative Medicine; Research; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Source; Spinal Cord; TCF Transcription Factor; Techniques; Tissues; Transcriptional Regulation; Transgenic Organisms; Undifferentiated; WNT Signaling Pathway; Work; Zebrafish; arm; cell behavior; cell motility; cell type; embryo cell; gastrulation; genome-wide analysis; human disease; improved; in vivo; insight; notochord; progenitor; stem cells; transcription factor; vertebrate embryos

Project Summary: We aim to understand the normal biology of neuromesodermal progenitors (NMPs), as well as to use NMPs to model cellular and molecular mechanisms in vivo. NMPs are basal progenitor cells located in the tailbud of vertebrate embryos that continue to make a germ-layer decision after gastrulation to generate ectoderm and mesoderm. As a primary source of cells generating the spinal cord, skeletal muscle, and other mesodermal derivatives, NMPs are a key cell type contributing to the formation of the vertebrate body plan. Studying NMPs has advanced our understanding of how body plans are generated, improved techniques for in vitro tissue generation, and provided critical insights into signaling pathway mechanisms. I discovered zebrafish NMPs as a postdoctoral fellow and my lab continues to focus on them. Our recent work has uncovered important roles for the transcription factor Sox2 in maintaining NMPs in an undifferentiated state through interactions with the canonical Wnt signaling pathway. This, combined with the past research showing the Wnt signaling effector bcat physically interacts with Sox2, indicates Wnt signaling can modulate gene expression and biological activity directly via Sox2. Little is known about this exciting branch of the Wnt/ bcat pathway. We will interrogate this pathway of Sox2 and Wnt/ bcat interactions at a number of biological levels, focusing on direct physical interactions between Sox2 and bcat, as well as genome wide analysis of Sox2/ bcat mediated transcriptional regulation. We hypothesize that Sox2/ bcat signaling represents a new arm of the Wnt/ bcat pathway distinct from the canonical TCF/LEF transcription factor family mediated signaling. We also recently developed a new Cyclin Dependent Kinase biosensor transgenic zebrafish and observed that NMPs and some of their derivatives exist in restricted cell cycle phases. The NMPs are held primarily in the G2 phase, while mesodermal notochord progenitors are restricted to the G1 phase. Cell cycle phase is broadly implicated in various aspects of stem cell maintenance, cell differentiation, and cell migration and invasion. We will manipulate the cell cycle in NMPs and their derivatives to understand how the cell cycle phase impacts normal NMP development. We hypothesize that the G2 phase restriction of NMPs is essential for maintenance of the undifferentiated state and for receipt of Wnt signaling based on G2 dependent receptor activation. We also hypothesize that the G1 phase of notochord progenitors is essential for their morphogenetic behavior of convergence and extension. Together, our work will shed important light on not only NMP development and vertebrate body plan formation, but also basic principles of cell biology and signaling. Wnt/ bcat signaling and Sox2 are found together and play important roles in numerous normal and diseased cellular contexts, including stem cells and cancer. The cell cycle is a fundamental aspect of normal development and is dysregulated in diseases such as cancer. Understanding how cell cycle phase impacts cell fate and morphogenetic behaviors in vivo will provide essential insight into normal and disease states.

项目概要： 我们的目标是了解神经中胚层祖细胞 (NMP) 的正常生物学，并利用 NMP 来 体内模型细胞和分子机制。 NMP 是位于尾芽的基底祖细胞 原肠胚形成后继续做出胚层决定以产生外胚层的脊椎动物胚胎 中胚层。作为产生脊髓、骨骼肌和其他中胚层细胞的主要来源 衍生物，NMP 是有助于脊椎动物身体计划形成的关键细胞类型。研究 NMP 增进了我们对身体计划如何生成的理解，改进了体外组织技术 的产生，并提供了对信号通路机制的重要见解。我发现斑马鱼 NMP 是 一名博士后研究员和我的实验室继续关注它们。我们最近的工作揭示了重要的角色 转录因子 Sox2 通过与 NMP 相互作用维持 NMP 处于未分化状态 经典Wnt信号通路。这与过去显示 Wnt 信号传导效应器的研究相结合 bcat 与 Sox2 发生物理相互作用，表明 Wnt 信号传导可以调节基因表达和生物学 直接通过 Sox2 进行活动。人们对 Wnt/bcat 通路的这个令人兴奋的分支知之甚少。我们将 在多个生物学水平上询问 Sox2 和 Wnt/bcat 相互作用的这条途径，重点关注直接 Sox2 和 bcat 之间的物理相互作用，以及 Sox2/ bcat 介导的全基因组分析 转录调控。我们假设 Sox2/ bcat 信号代表 Wnt/ bcat 的一个新分支 与典型的 TCF/LEF 转录因子家族介导的信号传导途径不同。我们最近也 开发了一种新的细胞周期蛋白依赖性激酶生物传感器转基因斑马鱼，并观察到 ​​NMP 和一些 它们的衍生物存在于受限的细胞周期阶段。 NMP 主要在 G2 阶段举行，而 中胚层脊索祖细胞仅限于 G1 期。细胞周期阶段广泛涉及 干细胞维持、细胞分化、细胞迁移和侵袭的各个方面。我们将 操纵 NMP 及其衍生物中的细胞周期，以了解细胞周期阶段如何影响正常 NMP开发。我们假设 NMP 的 G2 相限制对于维持 未分化状态并接收基于 G2 依赖性受体激活的 Wnt 信号传导。我们也 假设脊索祖细胞的 G1 期对其形态发生行为至关重要 收敛和延伸。我们的共同努力不仅将为 NMP 的开发和 脊椎动物身体计划的形成，也是细胞生物学和信号传导的基本原理。 Wnt/bcat 信号传导和 Sox2 一起被发现，并在许多正常和患病的细胞环境中发挥重要作用，包括 干细胞和癌症。细胞周期是正常发育的一个基本方面，并且在以下情况下失调： 癌症等疾病。了解细胞周期阶段如何影响细胞命运和形态发生行为 体内研究将为了解正常和疾病状态提供重要的见解。