CAREER: Optogel Proteomics: unbiased subcellular proteomics powered by photoreactions in hydrogel

将细胞想象成复杂的城市，蛋白质是城市的公民，每一个都在不同的地区扮演着特定的角色。为了让科学家了解这些细胞城市的功能，他们需要绘制出每个蛋白质公民的生活和工作地点。然而，这是一个相当大的挑战，因为目前的科学工具要么不够详细，要么一次只关注几个蛋白质。这个NSF职业项目介绍了一种名为光凝胶蛋白质组学的新方法，它就像一个高科技GPS，以惊人的精度跟踪比单个细胞还小的蛋白质。通过使用水凝胶和强大的显微镜，研究人员将从细胞内的准确位置分离和研究蛋白质。第一项任务是探索核层，这是一种包裹细胞细胞核的网络，可以因癌症或衰老而改变，以发现存在哪些蛋白质以及它们可能如何影响癌症的进展和衰老过程。该项目不仅旨在开创这项新技术，而且还将广泛分享，加强生物学和工程学方面的研究。此外，该项目的负责人是一名女科学家，她致力于通过特殊的夏令营和研讨会弥合科学领域的性别差距，赋予下一代女科学家权力。这项美国国家科学基金会的职业计划旨在通过发展光凝胶蛋白质组学来克服亚细胞空间蛋白质组学的局限性。通过使用膨胀显微镜、光反应和多光子显微镜，可以分离出精确的亚细胞位置的蛋白质，并随后通过质谱仪(MS)进行分析。该项目的目标是提供对亚细胞结构的无偏见、高分辨率的蛋白质组分析，最初的重点是核层，它与癌症相关和年龄相关的异染色质丢失有关。新的光凝胶蛋白质组学将与最新开发的超分辨率显微镜方法--标记保留扩展显微镜(LR-EXM)进行验证。更广泛的影响包括光凝胶蛋白质组学和LR-EXM的传播，以最低的成本提高生命科学的研究能力。首席研究员还将举办关于这些空间分析技术的年度研讨会，并促进性别平等和跨学科的科学暴露。这一奖项反映了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。

Imagine cells as complex cities, where proteins are the citizens, each with specific roles in different districts. For scientists to understand how these cellular cities function, they need to map where each protein citizen lives and works. However, this is quite a challenge because current scientific tools are either not detailed enough or only focus on a few proteins at a time. This NSF CAREER project introduces a new method named Optogel Proteomics, which acts like a high-tech GPS to track proteins with remarkable precision that is smaller than a single cell. By using hydrogels and a powerful microscope, the investigators will isolate and study proteins from exact locations within the cell. The first mission is to explore the nuclear lamina, a meshwork wrapping a cell's nucleus that can change due to cancer or aging, to discover what proteins are present and how they may influence cancer progression and the aging process. This project not only aims to pioneer this new technology but also to share it widely, enhancing research in biology and engineering. Additionally, the project’s leader, a woman scientist, is dedicated to bridging the gender gap in science through special camps and workshops, empowering the next generation of female scientists. This NSF CAREER project seeks to overcome the limitations of subcellular spatial proteomics through the development of Optogel Proteomics. By employing expansion microscopy, photoreactions, and multi-photon microscopy, proteins within precise subcellular locations can be isolated and subsequently analyzed via mass spectrometry (MS). The project's objective is to provide an unbiased, high-resolution proteomic analysis of subcellular structures, with an initial focus on the nuclear lamina, which has been implicated in cancer-related and age-related heterochromatin loss. The novel Optogel Proteomics will be validated against the newly developed super-resolution microscopy method, Label-Retention Expansion Microscopy (LR-ExM). The broader impacts include the dissemination of Optogel Proteomics and LR-ExM, enhancing research capacity in life sciences at minimal costs. The principal investigator will also conduct annual workshops on these spatial analysis technologies and promote gender equity and interdisciplinary science exposure.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.