Lineage-Specific Mechanisms of Cell Cycle Timing Control

细胞周期计时控制的谱系特异性机制

• 批准号: 10715965
• 负责人: Pavak Kirit Shah
• 金额: $ 33.81万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10715965
• 关键词: 3-Dimensional; Animals; Atlases; Biological Models; Caenorhabditis elegans; Cell Culture Techniques; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Fate Control; Cell Lineage; Cells; Collaborations; Dependence; Development; Developmental Process; Disease; Embryo; Evolution; Exhibits; Gene Transfer Techniques; Genetic; Homeostasis; Human; Image; In Vitro; Individual; Label; Link; Malignant Neoplasms; Microscopy; Modeling; Molecular; Molecular Analysis; Natural regeneration; Nematoda; Pattern; Recording of previous events; Regulation; Resources; Same-sex; Somatic Cell; Stereotyping; Structure; Study models; Techniques; Technology; Tissues; Work; cell fate specification; cell transformation; deep learning; design; gene network; insight; mutant; novel strategies; programs; reconstruction; stem cells; tool; transcriptomics

Project Summary Decades of studying animal development and in vitro human cell culture have produced many observed tight correlations between the duration of a cell’s cycle and its identity or the fates of its progeny. These links represent a unique opportunity to understand the regulatory relationships between genetic programs of cell fate and the regulation of the cell cycle, both central questions in the study of development, tissue homeostasis, regeneration, and proliferative disorders such as cancer. The nematode Caenorhabditis elegans has been a powerful model in which to study the regulation of cell fate and cell cycle control owing to its genetic tractability, transparent body and embryo, and stereotyped cell lineage. Like most nematodes, C. elegans exhibits eutely or a fixed number of somatic cells in each individual of the same sex. Cell fate in the wild-type animal can thus be determined solely on the basis of its lineage history, for which we have developed extensive tools and approaches for automated reconstruction via 3D timelapse microscopy. Using C. elegans and genetic perturbations that result in transformations of cell fate with its lineage, in combination with automated lineage tracing and spatial transcriptomics approaches, we will investigate the mechanisms by which cell fate influences the duration of a stem cell’s cell cycle as well as the mechanisms by which the duration of a cell cycle can influence cell fate. The work described in this proposal represents a novel approach to considering these links, enabled by our development of lineage tracing technologies and quantitative approaches to discovering structure in cell lineages. Building on this expertise, as well as our imaging resources and collaborations with other tools developers, theorists, and developmental biologists, we will continue to advance the state-of-the-art in lineage- resolved studies of metazoan development. In particular, using our advances in deep learning techniques to enable label-free automated lineage tracing in non-model species in which transgenesis remains impossible or difficult, we will leverage an evolutionary approach to understanding the design principles of gene networks that drive cell fate decisions and control cell cycle progression in the early embryo. Over the next five years we will complete detailed characterizations of co-dependencies between cell cycle timing and cell fate in the C. elegans embryo, create a molecular atlas of cell fate and cell cycle regulation in wild type and mutant C. elegans where cell fate patterning is perturbed, and complete the reconstruction and quantitative analysis of the embryonic lineages of S. stercoralis, P. pacificus, and C. angaria. In the long term, we plan to extend our molecular analyses to these species as well, beginning with C. angaria as an attractive model for studying the evolution of cell fate control networks and their interactions with regulators of the cell cycle. These insights will be of broad value to our understanding of developmental processes, and the resources we will establish will facilitate the work of others on diverse problems in emerging model systems.

项目摘要 数十年来研究动物发育和体外人类细胞培养已经产生了许多观察到的紧密 细胞周期的持续时间与其身份或进步的命运之间的相关性。这些链接 代表了了解细胞遗传程序之间的监管关系的独特机会 命运和细胞周期的调节，这两个中心问题在发育研究中，组织 体内稳态，再生和增殖疾病，例如癌症。线虫秀丽隐杆线虫 一直是一个强大的模型，用于研究细胞命运和细胞周期控制的调节 遗传障碍性，透明的身体和胚胎以及刻板印象的细胞谱系。像大多数线虫一样，C。 秀丽隐杆线虫在每个性别的每个个体中都表现出固定数量的体细胞。细胞命运 因此，野生型动物可以仅根据其谱系历史来确定，我们为此开发了 通过3D时间解体显微镜进行自动重建的广泛工具和方法。使用秀丽隐杆线虫 以及导致细胞命运及其谱系转化的遗传扰动，结合 自动谱系跟踪和空间转录组学方法，我们将通过 哪些细胞脂肪会影响干细胞细胞周期的持续时间以及 细胞周期的持续时间会影响细胞脂肪。 本提案中描述的工作代表了一种考虑这些链接的新颖方法，由我们启用 谱系追踪技术的开发和发现细胞中发现结构的定量方法 血统。以这些专业知识以及我们的成像资源和与其他工具的合作为基础 开发人员，理论家和开发生物学家，我们将继续推进血统的最先进 解析后生动物发展的研究。特别是，利用我们深入学习技术的进步 在非模型物种中启用无标签的自动谱系跟踪，其中转基因仍然不可能或 困难，我们将利用一种进化方法来理解基因网络的设计原理 这推动了细胞命运的决策和控制早期胚胎的细胞周期进程。在接下来的五年中 将在C中完全详细详细的依赖性特征。 秀丽隐杆线虫在野生型和突变体中创建一个细胞命运和细胞周期调节的分子图。 秀丽隐杆线虫在扰动细胞脂肪的情况下，并完成对的重建和定量分析 S. stercoralis，P。Pacificus和C. angaria的胚胎谱系。从长远来看，我们计划扩展我们的 分子分析也对这些物种进行了分析，首先是Anagria作为研究的有吸引力的模型 细胞命运控制网络的演变及其与细胞周期调节剂的相互作用。这些见解会 对于我们对发展过程的理解和我们将建立的资源具有广泛的价值 促进其他人对新兴模型系统中潜水员问题的工作。