Feedback between multiple signaling modes in control of a plant stem cell niche

控制植物干细胞生态位的多种信号传导模式之间的反馈

基本信息

批准号：
8894020
负责人：
ELLIOT M MEYEROWITZ
金额：
$ 31.3万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8894020
关键词：
Address Affect Affinity Chromatography Anabolism Angiosperms Animal Model Animals Apical Arabidopsis Attention Behavior Binding Biological Models Biology Cell Communication Cell Proliferation Cell division Cells ChIP-seq Chromatin Collection Communication Complex Computer Simulation Coupled Cytokinins Data Development Differentiated Gene Disease Encapsulated Epidermis Experimental Models Feedback Gene Expression Gene Expression Profile Gene Targeting Genes Genetic Genetic Transcription Genome Genomics Goals Growth and Development function Health Hormonal Hormones Hybrids Image Imaging Techniques In Vitro Laboratories Lead Libraries Life Location Maintenance Meristem Metabolism Methods Microscopy Modality Modeling Molecular Molecular Profiling Mouse-ear Cress Mutation Nuclear OmpR protein Organism Pathway interactions Peptide Signal Sequences Peptides Plant Shoots Plant Stems Plants Proteins Publishing RNA Regulation Regulator Genes Reporter Research Resolution Ribosomes Role Series Signal Pathway Signal Transduction Signaling Molecule Stem Cell Development Stem cells Sum Surface System Techniques Technology Testing Therapeutic Tissues Transgenes Transgenic Organisms Translating Yeasts cell behavior cell type comparative design feeding gain of function health application human disease human stem cells imaging modality in vivo intercellular communication mutant new technology novel novel strategies novel therapeutics organ regeneration overexpression receptor regenerative research study response screening stem cell biology stem cell niche stem cell population tissue regeneration transcription factor transcriptome sequencing yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Understanding the mechanism of maintenance and differentiation of stem cell populations is a central scientific issue on the path to realizing thefull therapeutic potential of stem cells in the treatment of human disease. Characterizing the cellular signals that maintain and regulate stem cell populations will lead to a new appreciation of the principles by which living organisms control growth and development and to the development of new technologies to utilize stem cells in health applications including tissue and organ regeneration. The proposed study, by characterizing the factors that regulate stem cell proliferation, differentiation, and specification in a model system, will reveal new principles of stem cell niche organization. The research proposed is designed to answer the central unanswered question in stem cell biology: what is the mechanism by which cell-cell communication instructs cellular behavior in stem cell populations? Because of its amenability to the full spectrum of methods from modern systems and genomic biology, as well as the remarkable regenerative abilities of plants, the shoot apical meristem (SAM) of the flowering plant Arabidopsis thaliana will be used as a model system. Two key signaling pathways, and their interactions, will be studied: the CLAVATA-WUSCHEL peptide signaling pathway and the hormonal cytokinin signaling pathway. The result of accomplishing the aims of the proposal will be a new level of understanding of stem cell niches, and the principles of feedback, signaling, and local cell type response that allow them to serve as permanent and regulated reservoirs of cells for creation of tissues. The experiments will lead to a new level of sophistication in our understanding of development, and of stem cell regulation in plants; and therefore to a more complete understanding of, and comparative view of, the principles of stem cell regulation in both plants and animals. Each aim will use non-invasive live imaging techniques that are particularly effective in plants, because their stem cells are near the surface, and transgenic reporter transgenes, as well as an extensive array of mutations that change the regulation of stem cell behavior. The goal of the first aim is a full characterization of the mechanisms by which alterations in the levels of the hormone cytokinin affect the different domains of the SAM stem cell niche, both directly, via regulation of the peptide signaling pathway, and by the multiple feedbacks of the hormone signaling system on itself. The second aim is a systems approach to characterize the cytokinin signaling pathway from receptor to response regulator transcription. Using the novel TRAP-seq and INTACT methods, cytokinin target genes will be characterized at cellular and molecular resolution. The third aim is an unbiased yeast-one hybrid screen to identify transcriptional regulators of cytokinin signaling, and cytokinin biosynthetic genes. The fourth and final aim is to use the data generated through these approaches to parameterize computational models that encapsulate the complex feedbacks in the system, to test new hypotheses of the mechanism by which cell-cell communication regulates the SAM stem cell niche.

描述（由申请人提供）：了解干细胞群体的维持和分化机制是实现干细胞在治疗人类疾病中的全部治疗潜力的核心科学问题。表征维持和调节干细胞群的细胞信号将导致对生物体控制生长和发育的原理有新的认识，并导致新技术的开发，以在包括组织和器官再生在内的健康应用中利用干细胞。拟议的研究通过表征模型系统中调节干细胞增殖、分化和规范的因素，将揭示干细胞生态位组织的新原理。拟议的研究旨在回答干细胞生物学中未解答的核心问题：细胞间通讯指导干细胞群中细胞行为的机制是什么？由于其适用于现代系统和基因组生物学的全方位方法，以及植物卓越的再生能力，开花植物拟南芥的茎尖分生组织（SAM）将被用作模型系统。将研究两个关键信号通路及其相互作用：CLAVATA-WUSCHEL 肽信号通路和激素细胞分裂素信号通路。实现该提案目标的结果将是对干细胞生态位以及反馈、信号传导和局部细胞类型反应原理的新水平的理解，使它们能够作为用于创建组织的永久和受调节的细胞库。这些实验将使我们对植物发育和干细胞调控的理解达到一个新的水平。因此，可以更全面地了解和比较植物和动物干细胞调节的原理。每个目标都将使用对植物特别有效的非侵入性实时成像技术，因为它们的干细胞靠近表面，以及转基因报告基因，以及改变干细胞行为调节的广泛突变。第一个目标的目标是全面表征激素细胞分裂素水平的变化影响 SAM 干细胞生态位不同域的机制，既通过肽信号传导途径的调节直接影响，又通过多重反馈影响激素信号系统自身的影响。第二个目标是一种系统方法来表征从受体到反应调节器转录的细胞分裂素信号传导途径。使用新型 TRAP-seq 和 INTACT 方法，细胞分裂素靶基因将以细胞和分子分辨率进行表征。第三个目标是进行无偏见的酵母一杂交筛选，以鉴定细胞分裂素信号传导和细胞分裂素生物合成基因的转录调节因子。第四个也是最后一个目标是使用通过这些方法生成的数据来参数化封装系统中复杂反馈的计算模型，以测试细胞间通讯调节 SAM 干细胞生态位的机制的新假设。