Notch-mediated modulation of Sonic hedgehog signaling in neural fate specification and differentiation

神经命运规范和分化中Notch介导的Sonic hedgehog信号传导调节

基本信息

批准号：
10223452
负责人：
BENNETT G NOVITCH
金额：
$ 19.25万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10223452
关键词：
Achievement Address Adopted Affect Automobile Driving Biological Phenomena Brain CRISPR/Cas technology Cell Maintenance Cells Chemicals Cilia Competence Complex Congenital Abnormality Data Data Set Defect Development Developmental Biology Disease Embryo Environment Exploratory/Developmental Grant Exposure to Fibroblasts Gene Expression Profiling Genes Genetic Transcription Goals Human Individual Instruction Ligands Maintenance Mammalian Cell Mediating Meta-Analysis Methods Movement Mus Mutation Nature Nervous System Trauma Nervous system structure Neuroglia Neurons Notch Signaling Pathway Organ Organism Outcome Pathway interactions Pattern Plant Roots Play Process Processed Genes Proteins Publishing Receptor Signaling Research Role SHH gene Shapes Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Skeletal Myoblasts Sonic Hedgehog Pathway Specific qualifier value Spinal Cord Structure Time Tissues Work body system cell fate specification cell type ciliopathy combinatorial developmental disease embryonic stem cell experimental study extracellular gain of function glial cell development hedgehog signal transduction in vivo insight morphogens nerve stem cell neurogenesis neuron development notch protein novel preservation prevent progenitor protein transport relating to nervous system repaired response smoothened signaling pathway sonic hedgehog receptor stem cells trafficking

项目摘要

PROJECT SUMMARY The formation of complex tissue and organs systems in developing organisms depends on the ability of dividing stem and progenitor cells to properly integrate extracellular signals present in the embryonic environment. The combined actions of these signals in turn direct diverse processes such as progenitor maintenance, differentiation, and assignment of specific cell fates. A key step towards understanding the basis of and means for preventing birth defect thus lies in defining how different signaling pathways mechanistically intersect, permitting the activation of one signaling pathway to influence responses to other signals. Moreover, the combinatorial activities have the potential to extend the range of outcomes possible from a limited range of developmental signals. Our studies in the developing spinal cord have identified an unexpected role for Notch receptor signaling in modulating the response of cells to the tissue morphogen Sonic hedgehog (Shh). Both activation and inactivation of the Notch pathway alters the dorsoventral register of neural progenitors, leading to corresponding changes in neuronal and glial cell fates (Kong et al. Dev Cell, 2015). In tracking down the mechanism behind these effects, we discovered that Notch signaling influences the trafficking of the Shh receptor Patched1 (Ptch1) and the key downstream Shh effector Smoothened (Smo) to primary cilia, leading to changes in downstream Shh pathway activities. Importantly, this role for Notch can be seen in multiple cell types including mouse and human neural progenitors, fibroblasts, and skeletal myoblasts, suggesting it may be a general feature of mammalian cells. Collectively, our studies reveal a novel and surprisingly proximal role for Notch in shaping the interpretation of the Shh morphogen gradient and thereby impacting cell fate determination. The means by which Notch influences the trafficking of Shh signaling proteins, however, remains unknown. Our preliminary data suggest that these actions of Notch are most likely transcriptionally mediated, raising the questions of what are the target genes regulated by Notch, and how do they impact the trafficking of Shh pathway components, and possibly other signaling proteins, to primary cilia? Moreover, do defects in this Notch-mediated pathway contribute to congenital defects affecting ciliary transport, collectively termed ciliopathies? Our proposed studies will address these questions, first by identifying the transcriptional targets of Notch and its downstream effector Hes1 that coincide with changes in ciliary trafficking, and second by developing a platform for investigating the function of these newly identified Notch target genes in Shh signaling through CRISPR/Cas9-mediated deletions. With this approach, we will: a) reveal the nature of the functional intersection between the Notch and Shh transduction pathways in neural fate selection and b) identify new regulators of Shh signaling and protein trafficking to primary cilia more generally.

项目摘要开发生物中复杂组织和器官系统的形成取决于将茎和祖细胞分开以正确整合胚胎中存在的细胞外信号环境。这些信号的综合动作反过来直接多样化的过程，例如祖先特定细胞命运的维持，分化和分配。理解基础的关键步骤因此，预防出生缺陷的方法和手段在于定义机械上不同的信号通路相交，允许激活一个信号通路以影响对其他信号的响应。而且，组合活动有可能从有限的范围扩大结果范围发展信号。我们在发育中的脊髓的研究确定了Notch的意外作用受体信号传导调节细胞对组织形态学声音刺猬（SHH）的反应。两个都 Notch途径的激活和失活改变了神经祖细胞的背腹式寄存器，领先神经元和神经胶质细胞命运的相应变化（Kong等人Dev Cell，2015）。在追踪这些效果背后的机制，我们发现Notch信号传导会影响SHH的运输受体修补1（PTCH1）和钥匙下游SHH效应器平滑（SMO）到原发性纤毛，导致下游SHH路径活动的变化。重要的是，在多个单元中可以看到Notch的角色包括小鼠和人类神经祖细胞，成纤维细胞和骨骼成肌细胞在内的类型，表明可能是哺乳动物细胞的一般特征。总的来说，我们的研究揭示了一个新颖而令人惊讶的近端作用在塑造SHH形态梯度的解释时缺口，从而影响细胞命运决心。但是，Notch影响SHH信号蛋白的运输的手段仍然未知。我们的初步数据表明，Notch的这些动作很可能是转录的介导的，提出了notch调节的目标基因的问题，以及它们如何影响将SHH途径成分以及可能其他信号蛋白的贩运运输到原发性纤毛？而且，做这种缺陷介导的途径中的缺陷导致了影响睫状运输的先天性缺陷被称为纤毛病？我们提出的研究将首先通过识别转录来解决这些问题 Notch及其下游效应子Hes1的目标与睫毛贩运的变化一致，第二通过开发一个平台来研究SHH中这些新鉴定的Notch靶基因的功能通过CRISPR/CAS9介导的缺失发出信号。使用这种方法，我们将：a）揭示神经命运选择和b中缺口和SHH转导途径之间的功能相交更普遍地将SHH信号传导和蛋白质运输的新调节剂确定为原发性纤毛。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

In vitro atlas of dorsal spinal interneurons reveals Wnt signaling as a critical regulator of progenitor expansion.

DOI：
10.1016/j.celrep.2022.111119
发表时间：
2022-07-19
期刊：
CELL REPORTS
影响因子：
8.8
作者：
Gupta, Sandeep;Kawaguchi, Riki;Heinrichs, Eric;Gallardo, Salena;Castellanos, Stephanie;Mandric, Igor;Novitch, Bennett G.;Butler, Samantha J.
通讯作者：
Butler, Samantha J.

Derivation of dorsal spinal sensory interneurons from human pluripotent stem cells.