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.

项目概要 数十年的动物发育和体外人类细胞培养研究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其后代的命运之间的相关性这些联系。 代表了了解细胞遗传程序之间的调控关系的独特机会 命运和细胞周期的调节，都是发育、组织研究的核心问题 体内平衡、再生和增殖性疾病，例如癌症。 已成为研究细胞命运调节和细胞周期控制的强大模型 遗传易处理性、透明的身体和胚胎以及定型的细胞谱系，与大多数线虫一样，C. 线虫在同一性别的每个个体中表现出相同或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，为此我们开发了 使用线虫通过 3D 延时显微镜进行自动重建的广泛工具和方法。 以及导致细胞命运及其谱系转变的遗传扰动，结合 自动谱系追踪和空间转录组学方法，我们将通过以下方式研究其机制 哪些细胞命运影响干细胞细胞周期的持续时间以及细胞命运的机制 细胞周期的持续时间可以影响细胞的命运。 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，由我们的支持 谱系追踪技术和发现细胞结构的定量方法的发展 建立在这种专业知识以及我们的成像资源和与其他工具的合作的基础上。 开发者、理论家和发育生物学家，我们将继续推进谱系的最先进水平- 特别是利用我们在深度学习技术方面的进步来解决后生动物发育的研究。 在转基因仍然不可能的非模型物种中实现无标记的自动谱系追踪，或者 困难，我们将利用进化方法来理解基因网络的设计原理 在接下来的五年里，我们将驱动细胞命运决定并控制早期胚胎的细胞周期进程。 将完成 C 中细胞周期时间和细胞命运之间相互依赖性的详细表征。 线虫胚胎，创建野生型和突变型线虫细胞命运和细胞周期调控的分子图谱。 细胞命运模式受到干扰的线虫，并完成了细胞命运模式的重建和定量分析 从长远来看，我们计划扩展 S. stercoralis、P. pacificus 和 C. angaria 的胚胎谱系。 对这些物种也进行了分子分析，首先从 C. angaria 作为研究 这些见解将揭示细胞命运控制网络的进化及其与细胞周期调节因子的相互作用。 对我们理解发展过程以及我们将建立的资源具有广泛的价值 促进其他人解决新兴模型系统中的各种问题的工作。