Investigations of proteome turnover kinetics under cellular differentiation

细胞分化下蛋白质组周转动力学的研究

• 批准号: 10705639
• 负责人: Edward Lau
• 金额: $ 38.88万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-17 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705639
• 关键词: Animal Model; Area; Biological; Cardiac Myocytes; Cell Differentiation process; Cells; Data Set; Deuterium; Deuterium Oxide; Disease Marker; Disease Progression; Disease model; Ensure; Event; Hepatocyte; Human; Investigation; Kinetics; Knowledge; Maps; Mass Spectrum Analysis; Methods; Modeling; Molecular; Physiological; Process; Production; Protein Biosynthesis; Proteins; Proteome; Research; Software Tools; Stable Isotope Labeling; Techniques; Tissue Engineering; Transcript; cell type; data tools; density; dynamic system; experimental study; induced pluripotent stem cell; kinetic model; knowledge of results; nerve stem cell; novel; protein degradation; stem cell differentiation; success

PROJECT SUMMARY Investigations of Proteome Turnover Kinetics Under Cellular Differentiation: Proteins are in a constant flux of continuous synthesis and degradation, both of which contribute to regulate protein levels in the cell. Although this fact has been known for 80 years, currently most biological inquiries are limited to static snapshots of overall transcript and protein levels, whereas knowledge into the dynamic changes of protein turnover remains severely lagging. This ESI MIRA proposal seeks to advance the current understanding of how cellular proteomes remodel during cell state transitions, by incorporating protein turnover kinetics information at key stages of human induced pluripotent stem cell (iPSC) differentiation into distinct cellular lineages. Working with collaborators, we previously developed deuterium stable isotope labeling, mass spectrometry, and kinetic modeling methods to quantitate protein turnover in animal models and in human. In doing so, we found many novel cell states and disease markers may be discovered from integrating orthogonal protein abundance and kinetics information. In the next five years, we propose to: (1) apply these methods to acquire a high temporal density map of human iPSC trilineage differentiation into cardiomyocyte, hepatocyte, and neuroprogenitor cells; (2) interrogate the regulatory principles that govern turnover flux across different differentiation stages; and (3) assess the functional consequences of protein degradation on the success and cell maturity of terminal cell production. Finally, a limitation of current techniques is that the kinetic models used in protein turnover studies largely assume a non-changing protein pool size at steady state, which does not apply to differentiating cells or progressing diseases. We propose to expand current models of analyzing heavy water stable isotope label experiments toward dynamical systems with variable protein pool sizes. If successful, the proposed research would greatly expand the current knowledge of molecular events that take place during human iPSC differentiation, as well as generate publicly available data sets and software tools to advance protein turnover studies in diverse areas.

项目总结 细胞分化条件下蛋白质组周转动力学研究：蛋白质处于恒流状态 持续的合成和降解，两者都有助于调节细胞中的蛋白质水平。虽然 这一事实已经知道80年了，目前大多数生物查询仅限于静态快照 整体转录本和蛋白质水平，而对蛋白质周转动态变化的知识仍然存在 严重滞后。这项ESI Mira提案旨在推进当前对细胞 蛋白质组在细胞状态转换过程中通过在关键位置加入蛋白质周转动力学信息来进行重构 人类诱导多能干细胞(IPSC)分化为不同细胞系的阶段。使用 合作者们，我们之前开发了稳定的氚同位素标记、质谱学和动力学 动物模型和人类中蛋白质周转量化的建模方法。在这样做的过程中，我们发现了 通过将正交蛋白质丰度和疾病标记相结合，可以发现新的细胞状态和疾病标记物 运动学信息。在接下来的五年里，我们建议：(1)应用这些方法来获得较高的时间 人IPSC三系分化为心肌细胞、肝细胞和神经前体细胞的密度图； (2)询问管理不同分化阶段的周转流量的监管原则；以及(3) 评估蛋白质降解对终末细胞成熟度和成熟度的影响 制作。最后，目前技术的局限性是，蛋白质周转研究中使用的动力学模型 在很大程度上假设蛋白质池大小在稳定状态下不变，这不适用于分化细胞 或进展中的疾病。我们建议扩展现有的重水稳定同位素标记分析模型 对具有可变蛋白质池大小的动力系统的实验。如果成功，拟议的研究 将极大地扩展目前关于人类IPSC期间发生的分子事件的知识 以及生成公开可用的数据集和软件工具以提高蛋白质周转 不同领域的研究。