Developmentally programmed remodeling of apical ECM

根尖 ECM 的发育程序重塑

• 批准号: 10344912
• 负责人: Maxwell Heiman
• 金额: $ 62.26万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10344912
• 关键词: Adolescent; Adult; Afferent Neurons; Animals; Apical; Attention; Biological Assay; Caenorhabditis elegans; Cell surface; Cells; Chemicals; Cilia; Complex; Cues; Data; Development; Diffuse; Disease; Environment; Epithelial; Epithelial Cells; Event; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Felis catus; Fluorescence; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Genetic Screening; Hermaphroditism; In Vitro; Lead; Life; Modeling; Morphogenesis; Neuroglia; Neurons; Neurotransmitters; Organ; Organism; Play; Process; Protein Family; Proteins; Regulatory Pathway; Role; Sense Organs; Sensory; Shapes; Structure; Surface; System; Testing; Therapeutic; Tissue Engineering; Tissues; Transducers; Tube; Work; base; cell type; density; differential expression; in vivo; innovation; male; mutant; nanoscale; novel; organ growth; prevent; programs; sex; targeted delivery; therapeutic target; tool; transcription factor

Abstract Apical extracellular matrix (aECM) coats the outward-facing surface of every organ, forming a barrier between the organism and its environment. Although it has been viewed historically as a static layer, aECM has been recently revealed to be dynamic across development and highly varied between cell types. It plays important roles in shaping organ morphogenesis and modulating cell activity. Identifying the regulatory mechanisms that control aECM composition and structure is therefore critical to understanding its role in development. Further, because aECM is highly accessible, there is enormous potential to manipulate it for targeted delivery of therapeutics or in tissue engineering. The major obstacles to studying aECM remodeling are that changes in aECM are difficult to visualize and need to be studied in vivo during highly dynamic processes that involve complex cell rearrangements. This project overcomes these obstacles by using an innovative model of developmentally programmed aECM remodeling. Preliminary data lead to the hypothesis that aECM structure is a discrete modular feature of cell identity, analogous to neurotransmitter types in neurons, rather than a continuum of stiffness/density. The C. elegans cuticle is an aECM that forms barrier between the animal and its environment. In order to directly access the external environment, the ciliated endings of some sensory neurons protrude through nanoscale pores in the cuticle, while those of other sensory neurons are embedded in specialized sheets of cuticle. Both types of cuticle structure are produced by glial cells that wrap the sensory neuron endings. One of these glial cells offers a remarkable example of developmentally programmed aECM remodeling: in juveniles of both sexes and in adult hermaphrodites it produces a cuticle sheet, but in adult males it produces a cuticle pore. This represents a discrete aECM remodeling event that occurs at a defined developmental stage without any major cell rearrangements. Preliminary data show that this aECM remodeling event is accompanied by a switch in gene expression in the glial cell. This includes expression of GRL-18, a novel class of aECM component that forms nanoscale rings in the cuticle. These observations leads to the hypothesis that a developmentally regulated switch in gene expression induces remodeling of aECM to form a nanoscale pore. To test this hypothesis, this project will (Aim 1) use mutant analysis and transcriptional profiling to define the gene expression switch that accompanies aECM remodeling; (Aim 2) determine how a novel developmentally regulated protein (GRL-18) contributes to aECM structure; and (Aim 3) test if changes in expression of GRL-18 and co-regulated genes are sufficient to remodel aECM into a nanoscale pore.

抽象的 顶端细胞外基质（AECM）覆盖每个器官的向外表面，形成障碍物 在生物体和环境之间。尽管历史上被视为静态层，但 最近已经揭示了AECM在整个开发过程中具有动态性，并且在细胞之间高度变化 类型。它在塑造器官形态发生和调节细胞活性方面起着重要作用。识别 因此，控制AECM组成和结构的调节机制对于 了解其在发展中的作用。此外，由于AECM高度可访问，因此有巨大的 操纵它的潜力，以靶向治疗药物或组织工程。专业 研究AECM重塑的障碍是，AECM的变化很难可视化，并且需要 在涉及复杂细胞重排的高度动态过程中，可以在体内研究。这 项目通过使用开发编程的AECM的创新模型来克服这些障碍 重塑。初步数据导致了以下假设：AECM结构是离散的模块化特征 细胞身份，类似于神经元中的神经递质类型，而不是刚度/密度的连续体。 秀丽隐杆线虫角质层是一种AECM，在动物及其环境之间形成障碍。在 为了直接访问外部环境，某些感觉神经元的纤毛结尾伸出 通过角质层中的纳米级孔，而其他感觉神经元的毛孔则嵌入 表皮的专业床单。两种类型的角质层结构都是由包裹的神经胶质细胞产生的 感觉神经元结尾。这些神经胶质细胞之一提供了一个非凡的开发例子 编程AECM重塑：在男女的少年中，在成年雌雄同体中，它会产生 角质层薄片，但在成年男性中会产生一个角质层孔。这代表离散的AECM重塑 事件发生在定义的发育阶段，而没有任何主要的细胞重排。初步的 数据表明，此AECM重塑事件伴随着基因表达的开关 细胞。这包括GRL-18的表达，GRL-18，这是一种形成纳米级环的新型AECM组件 在角质层中。这些观察结果导致了以下假设 基因表达诱导AECM的重塑形成纳米级孔。 为了检验该假设，该项目将（目标1）使用突变分析和转录分析到 定义AECM重塑伴随的基因表达开关； （目标2）确定小说如何 发育调节的蛋白质（GRL-18）有助于AECM结构。 （目标3）测试是否发生变化 在GRL-18和共同调节的基因的表达中，足以将AECM重塑为纳米级孔。