Understanding the robustness of cell cycles

项目摘要 生物振荡器对细胞和发育中的各种循环过程至关重要。其中包括Cell 分裂，心跳和体节发生受损的生物振荡器可能会导致失眠等疾病， 癌因此，对于振荡器来说，开发针对变化保持稳定功能的能力至关重要 in environmental环境conditions条件.许多振荡器的结构在物种之间是高度保守的， 实际的分子可能因物种而异。这就凸显了网络拓扑的重要作用 生物振荡器的功能。网络结构如何与生物的某些功能联系起来 振荡器仍然是系统和合成生物学中一个开放的挑战性问题。本提案的目的是 确定时钟网络稳健运行的基本原则。为了实现这一目标， 系统计算方法将被应用于分析所有拓扑修改， 影响时钟网络的鲁棒性和可调谐性。作为计算研究的比较，该建议 将通过实验研究细胞周期保持稳定振荡的可能机制。的 提出的实验利用基于液滴的微流体系统，其中无细胞提取物被 包封在液滴中以模拟经历有丝分裂周期的单细胞。这个人造细胞系统将 结合活胚胎成像和随机建模，跟踪和分析许多单个振荡器 同时，从而量化有丝分裂周期对环境变化的鲁棒性， 分子随机性为了研究网络结构在时钟鲁棒性中的作用， 振荡器将与其子网络受损的振荡器进行比较。有丝分裂的结果 时钟可以应用于共享类似拓扑核心的广泛的其他时钟集合。结果还应提供 关于如何设计一个强大的合成时钟的宝贵见解。

Project Summary Biological oscillators are essential to a variety of cyclic processes in cells and development. These include cell divisions, heartbeats, and somitogenesis. Impaired biological oscillators may cause diseases from insomnia to cancer. It is thus crucial for an oscillator to develop the ability to maintain a stable function against the changes in environmental conditions. The architecture of many oscillators is highly conserved among species, despite that the actual molecules may vary from species to species. This highlights the important role of network topology in the functions of biological oscillators. How the network structure is linked to the certain functions of biological oscillators is still an open challenging question in systems and synthetic biology. The goal of this proposal is to identify the fundamental principles underlying the robust functioning of clock networks. To achieve the goal, a systematic computational approach will be applied to analyze all topological modifications that significantly impact the robustness and tunability of clock networks. As a comparison to computational studies, this proposal will experimentally investigate the possible mechanisms by which cell cycles retain robust oscillations. The proposed experiments make use of a droplet-based microfluidic system, where cell-free extracts are encapsulated in droplets to mimic single cells that undergo mitotic cycles. This artificial cell system will be integrated with live embryo imaging and stochastic modeling, to track and analyze many single oscillators simultaneously, and thereby quantify the robustness of the mitotic cycles to environmental changes and molecular stochasticity. To study the role of network structure in the robustness of the clock, results from intact oscillators will be compared with the ones whose sub-networks are compromised. The results from the mitotic clock may apply to a broad set of other clocks that share similar topological cores. The results should also provide valuable insights on how to design a robust synthetic clock.