Characterizing dynamic properties and responses of an artificial mitotic cell

表征人工有丝分裂细胞的动态特性和响应

• 批准号: 1817909
• 负责人: Qiong Yang
• 金额: $ 67.5万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1817909&HistoricalAwards=false
• 关键词: Characterizing; dynamic; properties; responses; artificial

Mitosis is the important process by which cells divide and proliferate. A cell divides into two, and two into four, and so on. This process is finely controlled by a set of genes and proteins. These are known as the mitotic oscillator. Defects in mitosis are thought to increase the chance of developing cancer. It is difficult to study the oscillator inside the cell, where many other complicated cellular processes are also occurring. This project will build a synthetic mitotic oscillator into tiny droplets to mimic real cells. Without the complicated effects from cell growth and cell divisions, these microdroplets enable quantitative manipulation and characterization of the mitotic oscillator. The knowledge gained through this research will be disseminated to the broader scientific community through publications, conferences, and workshops. The interdisciplinary technologies will be disseminated to youth and the public through proposed outreach activities such as demonstrations and lab open day in collaboration with local professional educators and the museum. These activities will allow us to engage underrepresented minority students in grades 6-9 interested in science, expose them to research excitement, and prepare them for STEM careers.The goal of this project is to quantitatively analyze the stochastic dynamics of a minimal mitotic oscillator and its responses to various environmental signals. To constitute a minimal cytoplasmic mitotic oscillator, Xenopus egg cytosolic extracts will be encapsulated in microfluidic water-in-oil emulsions that mimic single cell behaviors. Additionally, a more complicated cell that contains nuclei will be built on top of this minimal oscillator. This will recapitulate complex phenomena in vitro such as nuclear assembly, chromatin condensation, and protein localization. Several questions will be examined in this system, including the role of cell size, temperature, and energy depletion in modulating properties of the oscillator. To achieve these goals, computational modeling, droplet-based microfluidic methods, and time-lapse fluorescence imaging will be integrated. This work will build a cyclic cell bottom-up, ranging from the simplest form containing no nuclei to the complicated ones driving various nuclear activities. The success of this work will provide valuable guidance in search of new targets for drug development and regenerative medicine in preventing and treating mitotic oscillator-related cancerous diseases, and thus be relevant and of great importance to broader communities in cancer biology, and cell and developmental biology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

有丝分裂是细胞分裂和增殖的重要过程。一个细胞分裂为两个，两个分裂为四个，等等，这个过程是由一组基因和蛋白质精细控制的。这些被称为有丝分裂振荡器。有丝分裂的缺陷被认为会增加患癌症的机会。很难研究细胞内的振荡器，因为许多其他复杂的细胞过程也在发生。这个项目将把一个合成的有丝分裂振荡器构建成微小的液滴来模仿真实的细胞。在没有细胞生长和细胞分裂的复杂影响的情况下，这些微滴使得有丝分裂振荡器的定量操作和表征成为可能。通过这项研究获得的知识将通过出版物、会议和讲习班传播给更广泛的科学界。跨学科技术将通过与当地专业教育工作者和博物馆合作的示范和实验室开放日等拟议的外联活动向青年和公众传播。这些活动将使我们能够吸引6-9年级对科学感兴趣的少数民族学生，让他们接触到研究的兴奋，并为STEM职业做好准备。本项目的目标是定量分析最小有丝分裂振荡器的随机动力学及其对各种环境信号的响应。为了构成最小的细胞质有丝分裂振荡器，将爪蟾卵胞质提取物封装在模拟单细胞行为的微流体油包水乳液中。此外，将在这个最小振荡器的基础上构建一个包含细胞核的更复杂的细胞。这将重演复杂的现象，在体外，如核组装，染色质凝聚，和蛋白质定位。几个问题将在这个系统中，包括细胞大小，温度和能量消耗的作用，在调制特性的振荡器进行检查。为了实现这些目标，将整合计算建模，基于液滴的微流体方法和延时荧光成像。这项工作将自下而上建立一个循环细胞，从不含核的最简单形式到驱动各种核活动的复杂形式。这项工作的成功将为药物开发和再生医学在预防和治疗有丝分裂增殖相关癌症疾病中寻找新靶点提供有价值的指导，因此对更广泛的癌症生物学社区具有重要意义。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值进行评估来支持和更广泛的影响审查标准。

项目成果

Building Dynamic Cellular Machineries in Droplet-Based Artificial Cells with Single-Droplet Tracking and Analysis

通过单液滴跟踪和分析在基于液滴的人造细胞中构建动态细胞机器

• DOI: 10.1021/acs.analchem.9b01481
• 发表时间: 2019
• 期刊: Analytical Chemistry
• 影响因子: 7.4
• 作者: Sun, Meng;Li, Zhengda;Wang, Shiyuan;Maryu, Gembu;Yang, Qiong
• 通讯作者: Yang, Qiong

Plug-in tubes allow tunable oil removal, droplet packing, and reaction incubation for time-controlled droplet-based assays

插入式管可实现可调节的除油、液滴堆积和反应孵育，以进行基于液滴的时间控制测定

• DOI: 10.1063/5.0047924
• 发表时间: 2021
• 期刊: Biomicrofluidics
• 影响因子: 3.2
• 作者: Sun, Meng;Maryu, Gembu;Wang, Shiyuan;Yang, Qiong;Bailey, Ryan C.
• 通讯作者: Bailey, Ryan C.