Effects of Mechanically-Induced Stress on the Proteome and Development

这笔拨款将支持更好地了解自然环境如何影响生物学的工作。具体来说，这笔拨款将支持更好地了解蛋白质如何对机械力作出反应的工作。生物体不断地感知机械力并作出适当的反应。例如，地球的重力不断向下拉，大气的压力从四面八方压缩，但生物可以保持直立。如果感知这些身体影响的能力受到损害，就会导致发育障碍、肌肉和骨质流失、神经异常、免疫和与年龄有关的问题。这些情况甚至在宇航员身上也能观察到，他们长时间远离地球，暴露在微重力环境中。在这项拨款下完成的工作使用了模拟不同物理环境的新型微技术系统。这些微技术系统将用于研究物理环境中的机械变化对数千种蛋白质的不同影响。确定这些生物反应的特征将最终改善国民健康状况。例如，这项工作的结果可能最终为未来的治疗策略或疾病预防技术提供信息。该项目还将创建一个跨学科合作的教育和培训计划。工程师和生物科学家将共同努力，并将扩大代表性不足群体参与研究的范围。地球上不同类型的物理力量不断地影响着生物，作为它们不断适应外部刺激的一部分。虽然对特定感觉系统中的机械转导了解很多，但对整个生物体对周围物理环境的广义机械感觉知之甚少。缺乏知识的一个原因是机械刺激足够的整个生物体以进行全面的蛋白质组学分析的方法有限。为了研究广义机械感觉，将使用即将开发的微制造高通量装置，将数百个活的果蝇胚胎暴露在多种模式、振幅和持续时间的各种机械刺激下，如微重力、超重力和外部压缩。这项工作将利用2D-DIGE（二维差异凝胶电泳）的比较蛋白质组学方法来识别蛋白质丰度或翻译后调节的变化，以响应这些不同的机械刺激模式。这种方法也将使局部/急性与慢性/无处不在的机械刺激的蛋白质组学比较成为可能。通过比较蛋白质组学与发育表型分析相结合，这些机械模式特化的程度，或在整个生物体背景下机械转导的普遍途径中重叠的程度将被检查。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This grant will support work to better understand how the physical environment influences biology. Specifically, this grant will support work to better understand how proteins respond to mechanical forces. Organisms are continuously sensing mechanical forces and respond appropriately. For example, the Earth’s gravity is constantly pulling down and the atmospheric pressure which compresses from all directions, yet organisms can remain upright. If the ability to sense these physical effects is compromised, it can lead to developmental disorders, muscle and bone loss, neurological abnormalities, immunological, and age-related problems. These conditions have been observed even in astronauts, who are exposed to microgravity during prolonged times away from Earth. The work done under this grant use novel microtechnology systems that mimic different physical environments. These microtechnology systems will be used to investigate how mechanical changes in the physical environment differentially affect thousands of proteins. Characterizing these biological responses will ultimately lead to improvements in national health. For example, the results of this work may ultimately inform future therapeutic strategies or disease prevention techniques. This project will also create an educational and training program for interdisciplinary collaboration. Engineers and biological scientists will work together, and participation in research for underrepresented groups will be broadened. Different types of Earth’s physical forces continuously impact living beings as a part of their ceaseless adaptation to external stimuli. While much is known about mechanotransduction in specific sensory systems, little is known about generalized mechanosensation of the surrounding physical environment by whole organisms. One reason for this lack of knowledge is the limited methods to mechanically stimulate enough whole organisms to enable comprehensive proteomic analysis. To investigate generalized mechanosensation, hundreds of living Drosophila embryos will be exposed to multiple modes, amplitudes, and durations of various mechanical stimulation such as microgravity, hypergravity, and external compression using microfabricated high-throughput devices that will be developed. This work will utilize the comparative proteomics approach of 2D-DIGE (2 Dimensional Difference Gel Electrophoresis) to identify changes in protein abundance or post-translational regulation in response to these different modes of mechanical stimulation. This approach will also enable the proteomic comparison of local/acute to chronic/ubiquitous mechanical stimulation. By using comparative proteomics in conjunction with the analysis of developmental phenotypes, the extent to which these mechanical modes are specialized, or overlap in a universal pathway of mechanotransduction in the context of whole organisms will be examined.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.