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.

项目摘要 在开发过程中，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 扩散或分化的能力最常丢失，因此大多数成年细胞通常无能力自我 再生或维修。再生疗法的发展将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 静态祖细胞行为的构建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。该提议的目的是 表征控制静止，分化，可能重要的基因的调节和功能 海洋无脊椎动物和脊椎动物ciona robusta的扩散。西奥纳是我们最接近的无脊椎动物 亲戚，因此在开发过程中，我们具有相似的结构和分子特征。但是，西奥纳 当幼虫身体计划时，从幼虫到成人形式进行了戏剧性的转换 退化并由静态的祖细胞代替，这些祖细胞必须绕过编程的细胞死亡才能重新出现 术后并产生成人身体计划。这样的细胞人群是幼虫神经祖细胞 称为颈部细胞，在离散的干细胞小众样室中建立。信号通路 直接进入颈部，维持和从静止的遗传成分仍然没有表征。 我建议使用颈部细胞种群作为模型来识别调节静止和的独特机制 可以用于未来疗法的再生。该提议的理由是，通过利用 Ciona的障碍性，这些颈部细胞的可及性及其刻板印象的细胞行为，我可以紧密 检查颈部细胞静止，增殖和分化的调节控制。我会追求两个 具体目标。 1）研究在幼虫阶段对颈部静止和增殖的控制 细胞外提示和细胞内转录对照的整合。 2）研究一种新的机制 转录启动和颈部细胞分化的延迟。我将使用创新来追求这些目标 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。拟议工作的预期结果包括识别先前未认可的功能 对于神经发育中的保守但研究不足的基因，以及它们的时空调节如何是 对静止的精确时机有启发性。这将为针对性调查的基础 神经发育过程是广泛的人类疾病。