BBSRC-NSF/BIO: Synthetic Control of Pattern Formation and Morphogenesis in a Purposefully Rewired Vertebrate Cell

这个项目的目标是合成(人工)控制细胞“皮质”的行为--细胞的最外层。正常情况下，大脑皮质为许多基本的生物过程提供动力，无论是在单个细胞内，还是在组织和器官中。该项目的完成将带来四个重要的研究成果：第一，它将测试目前关于生命系统如何执行细胞分裂、细胞运动和细胞形状变化等过程的想法。其次，它将提供新的工具和技术，允许操纵活着的有机体中的细胞行为。第三，它将提供促进新的、潜在有益的细胞行为的手段。第四，它将导致新的计算方法的发展，用于分析和理解复杂的细胞行为。该项目的成功完成还将产生若干重要的教育和培训成果。该项目是美国威斯康星大学麦迪逊分校和英国爱丁堡大学研究人员的合作项目，将支持两名博士后研究人员和一名研究生在细胞、分子和计算生物学方面的培训，进一步为他们在科学领域的职业生涯做好准备。此外，另有4-6名本科生将接受细胞和分子生物学方面的培训，为他们在科学或医学领域的职业生涯做好准备。最后，四名代表不足、经济资源不足的高中生将接受连续两个暑假的细胞和分子生物学培训，并接受如何在大学取得成功的额外培训。预计培训将为这些学生提供在科学、技术、工程和医学相关专业中脱颖而出所需的分析和定量技能，并可能最终在这些领域取得职业生涯。细胞皮质负责以适当的机械行为对各种内部和外部信号做出反应。这些行为包括细胞分裂、细胞运动和短期或长期的细胞形状变化。这个团队和其他人最近发现了一种动态过程--皮质兴奋性--各种类型的细胞利用这种过程来驱动不同的机械行为。大脑皮层的兴奋性外在表现为肌动蛋白组装的皮质波和控制肌动蛋白组装的各种大分子的互补波。皮层兴奋性本身受快速正反馈和延迟负反馈的共同控制。我们将开发在正常情况下不显示皮质兴奋性的细胞(即青蛙卵母细胞)中合成诱导皮质兴奋性的方法，并使用高分辨率活细胞成像来捕捉兴奋性的详细特征。这种诱导将基于人工合成的蛋白质结构，这种结构可以产生快速的正反馈，也可以产生延迟的负反馈。这些结构将能够产生不同的皮质兴奋性机制，要么是全局的(即整个皮质)，要么是局部的(即皮质的不同区域或模式)。通过组合不同的合成结构，我们将驱动简单的细胞形状变化(如沟槽)或复杂的细胞形状变化(如原肠形成)，使我们能够测试细胞形状控制的基本想法。此外，通过迭代地将实验与结果的计算建模相结合，将有可能发展对细胞分裂和形态发生等过程的定量的、机械性的理解。从细胞和发育生物学到计算建模，再到物理学，从事各种科学学科的人都会对这项研究感兴趣。此外，由于生成的数据将非常丰富的信息，我们预计它将成为许多其他对动态行为感兴趣的研究人员的资源。这一美英合作项目得到了美国国家科学基金会和英国生物技术和生物科学研究委员会的支持。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The goal of this project is to synthetically (artificially) control the behavior of the cell "cortex"—the outermost layer of the cell. It is the cortex that normally powers many fundamental biological processes, both within single cells, and in tissues and organs. Completion of the project will result in four important research outcomes: first, it will test current ideas about how living systems execute such processes as cell division, cell movement, and cell shape change. Second, it will provide new tools and technologies that permit manipulation of cell behavior in living organisms. Third, it will provide the means to promote new, and potentially beneficial cell behaviors. Fourth, it will result in the development of new computational approaches for the analysis and understanding of complex cell behaviors. Successful completion of the project will also lead to several important educational and training outcomes. This project, which is a collaboration between researchers at the University of Wisconsin-Madison (US) and the University of Edinburgh (UK), will support the training of two postdoctoral researchers and one graduate student in cell, molecular and computational biology, further preparing them for careers in science. Moreover, an additional four-six undergraduate students will be trained in cell and molecular biology, preparing them for careers in science or medicine. Finally, four under-represented and financially underresourced high school students will be trained in cell and molecular biology for two consecutive summers and provided with additional training on how to succeed in college. The training is anticipated to provide these students with the analytical and quantitative skills needed to excel in science, technology, engineering, and medicine related majors and, presumably, eventual careers in these areas.The cell cortex is responsible for responding to a variety of internal and external signals with the appropriate mechanical behavior. Such behaviors include cell division, cell locomotion and short or long-term cell shape changes. This team and others recently discovered a dynamical process—cortical excitability—that a variety of cell types harness to drive distinct mechanical behaviors. Cortical excitability is outwardly manifest as propagating cortical waves of actin assembly and complementary waves of the various macromolecules that control actin assembly. Cortical excitability is itself controlled by coupled fast positive feedback and delayed negative feedback. We will develop the means to synthetically induce cortical excitability in cells that do not normally display it, namely, frog oocytes, and employ high-resolution live cell imaging to capture the detailed features of excitability. The induction will be based on synthetic protein constructs engineered to produce either fast positive feedback or delayed negative feedback. These constructs will be capable of generating different cortical excitability regimes either globally (ie throughout the entire cortex) or locally (ie in distinct regions of or patterns in the cortex). By combining different synthetic constructs, we will drive simple cell shape changes (ie furrowing) or complex cell shape changes (ie gastrulation), allowing us to test basic ideas about cell shape control. In addition, by iteratively combining experiments with computational modeling of the results, it will be possible to develop both a quantitative, mechanistic understanding of processes such as cell division and morphogenesis. This research will be of interest to those working in a broad variety of scientific disciplines, ranging from cell and developmental biology to computational modeling, to physics. Moreover, because the data generated will be extraordinarily rich in information, we anticipate that it will serve as a resource for many other researchers interested in dynamical behavior.This collaborative US/UK project is supported by the US National Science Foundation and the UK Biotechnology and Biological Sciences Research Council.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.