Dissecting gene regulation of stem cell quiescence in Ciona

剖析玻璃海鞘干细胞静止的基因调控

• 批准号: 10679206
• 负责人: Eduardo D. Gigante
• 金额: $ 7.2万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679206
• 关键词: Adolescent; Adult; Apoptosis; Behavior; Biological Assay; Biological Metamorphosis; Brain; Bypass; C-terminal; CRISPR/Cas technology; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cells; Cephalic; Chordata; Cues; Data; Development; Developmental Biology; Disease; Embryo; Equilibrium; Fluorescence Microscopy; Foundations; Future; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Gills; Goals; Homeobox; Homologous Gene; Human; Individual; Invertebrates; Investigation; Knock-out; Larva; Life; Link; Maintenance; Mammals; Marine Invertebrates; Mediating; Modeling; Molecular; Motor Neurons; Muscle; Natural regeneration; Neck; Nervous System; Neuronal Differentiation; Neurons; Organ; Organism; Outcome; Pathway interactions; Pattern; Pharyngeal structure; Play; Population; Process; Proliferating; RNA Polymerase II; Regenerative capacity; Regulation; Regulator Genes; Regulatory Element; Reporter; Role; Signal Pathway; Signal Transduction; Sister; Specific qualifier value; Stereotyping; Structure; Time; Tissues; Transcription Elongation; Transcriptional Regulation; Urochordata; Vertebrates; Work; antagonist; ascidian; bone; brain cell; cell behavior; cholinergic neuron; extracellular; genetic architecture; genome editing; in vivo Model; innovation; knockout gene; mutant; nerve stem cell; nervous system development; neurodevelopment; new technology; novel; prevent; progenitor; receptor; regeneration potential; regenerative; regenerative biology; regenerative therapy; repaired; response; spatiotemporal; stem cell niche; stem cell population; stem cells

PROJECT SUMMARY During development, the formation of heterogenous tissues, organs, and cellular networks depends on a careful balance between cellular proliferation, quiescence, and differentiation. Once cells have fully differentiated, their ability to proliferate or differentiate are most often lost, and so most adult cells are generally incapable of self- regeneration or repair. The development of regenerative therapies will require harnessing this latent ability, but first we need a deeper understanding of the processes that control it. Thus, understanding the genetic architecture underlying quiescent progenitor behavior is key to developing and applying emerging therapies with new technologies such as cellular reprogramming or genome editing. The objective of this proposal is to characterize the regulation and functions of potentially important genes controlling quiescence, differentiation, and proliferation in marine invertebrate and chordate Ciona robusta. Ciona are among our closest invertebrate relatives, and so during development we share similar structural and molecular features. However, Ciona undergo a dramatic conversion from larval to adult forms called metamorphosis, when the larval body plan degenerates and is replaced by quiescent progenitors which must bypass programmed cell death to reemerge post-metamorphosis and generate the adult body plan. One such cell population are larval neural progenitors called Neck cells which are established in a discrete stem cell niche-like compartment. The signaling pathways and genetic components that direct Neck entry, maintenance, and exit from quiescence remain uncharacterized. I propose using the Neck cell population as a model to identify unique mechanisms regulating quiescence and regeneration that can be harnessed for future therapies. The rationale for this proposal is that, by exploiting the tractability of Ciona, the accessibility of these Neck cells, and their stereotyped cellular behaviors, I can closely examine regulatory control of Neck cell quiescence, proliferation, and differentiation. I will do so by pursuing two specific aims. 1) To investigate the control of Neck quiescence and proliferation during the larval stage by the integration of extracellular cues and intracellular transcriptional control. 2) To investigate a novel mechanism for transcriptional priming and delay of Neck cell differentiation. I will pursue these aims using an innovative approach that combines cell lineage-specific, CRISPR/Cas9-based somatic gene knockouts and fluorescence microscopy. The expected outcomes of the proposed work include identifying previously unrecognized functions for conserved but poorly studied genes in neurodevelopment and how their spatiotemporal regulation can be instructive for the precise timing of quiescence. This will establish a foundation for a targeted investigation of neurodevelopmental processes underlying a wide range of human disorders.

项目概要 在发育过程中，异质组织、器官和细胞网络的形成取决于仔细的 细胞增殖、静止和分化之间的平衡。一旦细胞完全分化，它们的 增殖或分化的能力通常会丧失，因此大多数成体细胞通常无法自我增殖或分化。 再生或修复。再生疗法的发展需要利用这种潜在能力，但是 首先，我们需要更深入地了解控制它的过程。因此，了解遗传 静态祖细胞行为的基础结构是开发和应用新兴疗法的关键 新技术，例如细胞重编程或基因组编辑。该提案的目的是 表征控制静止、分化的潜在重要基因的调控和功能， 以及海洋无脊椎动物和脊索动物海鞘的增殖。 Ciona 是与我们最亲近的无脊椎动物之一 亲戚，因此在发育过程中我们具有相似的结构和分子特征。然而，西奥娜 当幼虫身体规划时，经历从幼虫到成虫形式的戏剧性转变，称为变态 退化并被静止的祖细胞取代，这些祖细胞必须绕过程序性细胞死亡才能重新出现 变态后并生成成人身体计划。其中一种细胞群是幼虫神经祖细胞 称为颈部细胞，它们建立在离散的干细胞生态位样区室中。信号通路 指导颈部进入、维持和退出静止状态的遗传成分仍然未知。 我建议使用颈部细胞群作为模型来识别调节静止和 可用于未来治疗的再生。该提案的基本原理是，通过利用 Ciona 的易处理性、这些颈部细胞的可及性以及它们的刻板细胞行为，我可以密切关注 检查颈细胞静止、增殖和分化的调节控制。我将通过追求两个来做到这一点 具体目标。 1) 研究幼虫阶段颈部静止和增殖的控制 细胞外线索和细胞内转录控制的整合。 2）研究新机制 颈细胞分化的转录启动和延迟。我将通过创新的方式来实现这些目标 结合细胞谱系特异性、基于 CRISPR/Cas9 的体细胞基因敲除和荧光的方法 显微镜。拟议工作的预期成果包括确定以前未认识到的功能 神经发育中保守但研究很少的基因以及它们的时空调节如何 对于精确的静止时间具有指导意义。这将为有针对性的调查奠定基础 多种人类疾病背后的神经发育过程。