Extended super-resolution three-dimensional mechanical probing in living cells.

活细胞中的扩展超分辨率三维机械探测。

• 批准号: EP/S004459/1
• 负责人: Marco Fritzsche
• 金额: $ 54.36万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FS004459%2F1
• 关键词: Extended; super; resolution; three; dimensional

New perspective of mechanobiology is currently emerging across multiple disciplines in the physical and biomedical research fields. In contrast to conventional beliefs, recent evidence indicates that cells regulate their cell mechanics not only downstream of signalling events triggered by external stimuli, but that cells employ a diversity of feedback mechanisms of their cytoskeleton enabling them to dynamically adjust cell mechanics to meet physiological needs. Consequently, this provides a previously unforeseen picture wherein cells actively exert and resist biomechanical force to tune their mechanobiology, and thus facilitate their function. Quantifying cellular forces has therefore become an important mission across multiple disciplines at the interface of bioengineering and biomedical sciences. The overall goal of this project is the development of a new-generation force probing methodology to enable physiological mechanical probing in living cells at unprecedented accuracy and resolution. To engineer this technology, we will combine a new technique of state-of-the-art 3D high-speed total-internal-reflection (TIRF) and cutting-edge super-resolution structural-illumination (SIM) microscopy coupled with extended mechanical force TFM probing to create a novel breakthrough technology for mechanical probing in living cells. We will demonstrate the power of the new methodology by quantifying mechanical force production in a variety of adherent cells and activating immune T cells. We anticipate that our research might lead to the replacement of conventional TFM measurements with major implications for the understanding of the cellular mechanobiology. Ultimately, we therefore aim to commercialise the 3D eTIRF-SIM-TFM method.

机械生物学的新视角正在跨越物理和生物医学研究领域的多个学科。与传统观点相反，最近的证据表明，细胞不仅在外部刺激触发的信号事件下游调节其细胞力学，而且细胞还利用细胞骨架的多种反馈机制，使它们能够动态调节细胞力学以满足生理需要。因此，这提供了一个以前无法预见的画面，其中细胞主动施加和抵抗生物力学力来调整其机械生物学，从而促进其功能。因此，细胞力的量化已成为生物工程和生物医学交叉领域的一项重要任务。该项目的总体目标是开发新一代力探测方法，以前所未有的精度和分辨率在活细胞中进行生理机械探测。为了设计这项技术，我们将结合最先进的3D高速全内反射（TIRF）新技术和尖端的超分辨率结构照明（SIM）显微镜，再加上扩展的机械力TFM探测，为活细胞的机械探测创造一种新的突破性技术。我们将通过量化各种贴壁细胞和激活免疫T细胞的机械力产生来证明新方法的力量。我们预计我们的研究可能会导致取代传统的TFM测量，对细胞力学生物学的理解产生重大影响。最终，我们的目标是将3D eTIRF-SIM-TFM方法商业化。

项目成果

Semaphorin 3A induces cytoskeletal paralysis in tumor-specific CD8+ T cells

• DOI: 10.1101/849083
• 发表时间: 2019-11
• 期刊: bioRxiv
• 作者: M. Barnkob;Y. Michaels;Violaine Andre;P. Macklin;U. Gileadi;Salvatore Valvo;Margarida Rei;Corinna A. Kulicke;Ji-Li Chen;V. Jain;V. Woodcock;Huw Colin-York;A. Hadjinicolaou;Y. Kong;V. Mayya;J. Bull;P. Rijal;C. Pugh;A. Townsend;L. Olsen;M. Fritzsche;T. Fulga;Michael Loran Dustin;E. Jones;V. Cerundolo

Two-dimensional TIRF-SIM-traction force microscopy (2D TIRF-SIM-TFM).

二维 TIRF-SIM-牵引力显微镜 (2D TIRF-SIM-TFM)

• DOI: 10.1038/s41467-021-22377-9
• 发表时间: 2021-04-12
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Barbieri L;Colin-York H;Korobchevskaya K;Li D;Wolfson DL;Karedla N;Schneider F;Ahluwalia BS;Seternes T;Dalmo RA;Dustin ML;Li D;Fritzsche M
• 通讯作者: Fritzsche M

Sterile activation of invariant natural killer T cells by ER-stressed antigen-presenting cells

• DOI: 10.1073/pnas.1910097116
• 发表时间: 2019-11-19
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Bedard, Melissa;Shrestha, Dilip;Cerundolo, Vincenzo
• 通讯作者: Cerundolo, Vincenzo

Mechanobiological Control of the Immune Response

免疫反应的机械生物学控制

• DOI: 10.1016/j.bpj.2018.11.2959
• 发表时间: 2019
• 期刊: Biophysical Journal
• 影响因子: 3.4
• 作者: Colin-York H