Transduction of Substrate Mechanical Cues via Piezo1 Ion Channels

通过 Piezo1 离子通道转导基底机械信号

• 批准号: 2015964
• 负责人: Zonglu Susan Hua
• 金额: $ 39.05万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2015964&HistoricalAwards=false
• 关键词: Transduction; Substrate; Mechanical; Cues; via

This award will advance our fundamental understanding of how cells sense and react to diverse mechanical inputs from their environments. Cells utilize local environmental cues from the extracellular matrix (ECM), adhesive molecules, and local stiffness to make decisions about growth and movement. The process of converting mechanical stimulus to electrical energy at the ECM is commonly thought to act via adhesion proteins such as integrins. Emerging research on Piezo ion channels reveals that these channels can also respond to changes in the substrate mechanical properties. Therefore, these channels may serve as additional force sensors. This work will explore the role of a novel cell sensor, called Piezo1, in detecting ECM forces. The results may ultimately impact on human health. For example, the body’s mishandling of these mechanical signals results in a variety of diseases, including chronic kidney disease and cancer. The results of this work may lead to new types of treatment for these conditions. The outcome has the potential to be broadly applicable, since Piezo1 is found in many cell types and organs where the mechanical environment plays a major role in proper function. The research findings will be integrated into outreach activities through an Education through Experimentation (E2E) platform that involves graduate, undergraduate, and local high school students in this project.The project is based on the hypothesis that Piezo1 is the primary force sensor in epithelial cells, and that it detects the substrate mechanical cues translated into membrane tension. This occurs through a feedforward mechanism: local tension activates Piezo1 channels and calcium influx, which stimulates calcium-dependent contractility and cell spreading; the changes in cell geometry in turn, promote more Piezo1 activity and the migration of Piezo1 to regions under stress, causing further growth in the same direction. This hypothesis will be tested with substrate modulation as mechanical stimulus, a FRET-based fluorescent probe to report opening transitions in Piezo1, and a wound healing assay to screen cell mobility. There are three primary objectives. First, to identify ECM force sensors in cells by inhibiting Piezo1 activity and by knockdown of Piezo1. Second, to investigate how substrates alter Piezo1 responsiveness by visualizing the location and opening of channels during cell reshaping. Third, to test the hypothesis that functional Piezo1 expression level determines the identity of leader cells guiding collective cell migration. Taken together, the results of this work have the potential to transform our understanding of the mechanisms of various disease states that affect epithelial cells.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.

这个奖项将推进我们对细胞如何感知和反应来自环境的各种机械输入的基本理解。细胞利用来自细胞外基质（ECM）、粘附分子和局部刚度的局部环境信号来决定生长和运动。在ECM将机械刺激转化为电能的过程通常被认为是通过粘附蛋白（如整合素）起作用的。对压电离子通道的新研究表明，这些通道也可以响应衬底力学性能的变化。因此，这些通道可以作为额外的力传感器。这项工作将探索一种名为Piezo1的新型细胞传感器在检测ECM力方面的作用。其结果可能最终影响人类健康。例如，身体对这些机械信号的处理不当会导致各种疾病，包括慢性肾病和癌症。这项工作的结果可能会导致治疗这些疾病的新类型。该结果具有广泛应用的潜力，因为Piezo1存在于许多细胞类型和器官中，其中机械环境在正常功能中起主要作用。研究成果将通过实验教育（E2E）平台整合到推广活动中，参与该项目的研究生、本科生和当地高中生都将参与其中。该项目基于一个假设，即Piezo1是上皮细胞中的主要力传感器，它可以检测转化为膜张力的基质机械信号。这是通过前馈机制发生的：局部张力激活Piezo1通道和钙内流，从而刺激钙依赖性收缩和细胞扩散；细胞几何形状的变化反过来又促进了更多的Piezo1活性和Piezo1向应力区域的迁移，从而导致在同一方向上进一步生长。这一假设将通过底物调制作为机械刺激，基于fret的荧光探针来报告Piezo1的开放转变，以及伤口愈合试验来筛选细胞流动性来验证。有三个主要目标。首先，通过抑制Piezo1活性和敲低Piezo1来识别细胞中的ECM力传感器。其次，研究基底如何通过可视化细胞重塑过程中通道的位置和开放来改变Piezo1的响应性。第三，验证功能性Piezo1表达水平决定引导集体细胞迁移的领导细胞身份的假设。综上所述，这项工作的结果有可能改变我们对影响上皮细胞的各种疾病状态机制的理解。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。

项目成果

LOCAL ECM STIFFNESS MODULATES EPITHELIAL CELL RESPONSE TO MICROPATTERNS

局部 ECM 硬度调节上皮细胞对微图案的反应

• 发表时间: 2023
• 期刊: SB3C2023 Proceedings Book
• 作者: Tasnim Shireen, Rajath D.Prabhu

Contribution of Piezo1 in ECM stiffness incited epithelial cell remodeling

Piezo1 在 ECM 硬度刺激上皮细胞重塑中的贡献

• 发表时间: 2022
• 期刊: IMECE2022
• 作者: D. Jetta, Tasnim Shireen

Physical memory of astrocytes

星形胶质细胞的物理记忆

• DOI: 10.1016/j.brainres.2022.148076
• 发表时间: 2022
• 期刊: Brain Research
• 影响因子: 2.9
• 作者: Shireen, Tasnim;Sachs, Frederick;Hua, Susan Z.
• 通讯作者: Hua, Susan Z.

Piezo1 Is Essential for Cell Remodeling on Micropatterns

Piezo1 对于微图案的细胞重塑至关重要

• DOI: 10.1016/j.bpj.2020.11.1554
• 发表时间: 2021
• 期刊: Biophysical Journal
• 影响因子: 3.4
• 作者: Reza Bahrani Fard, Mohammad;Jetta, Deekshitha;Munechika, Katie;Hua, Susan Z.
• 通讯作者: Hua, Susan Z.