Piezo1机械力敏感离子通道的定量生物学和合成生物学研究

合成生物学利用工程化的思想，改造设计生物元件，构建合成基因线路，对细胞功能进行优化编程，使其能够按设计需求处理信息和行使功能，从而实现抵抗疾病和改善人类健康的目的，现已取得重大的基础和应用研究突破。然而力学感应这个生命过程中至关重要的信号系统，至今尚未被合成生物学成功改造。本项目通过对机械力敏感离子通道Piezo1的系统、定量刻画，研究其与外源表达的底盘细胞在静态和动态外力下的感知机制。通过运用合成生物学工具降低Piezo1外源表达的异质性，我们发现其开启只限于静态剪切力的特定强度以及音频振动的特定频率。在对其进行合成生物学改造前，我们也将对高强度剪切力脱敏和底盘细胞骨架对频率滤波的科学问题进行进一步的研究。为此我们将发展对细胞膜或细胞骨架直接施加高频力学扰动的技术，也将检测Piezo1的力学感知能够识别发生转移的肺腺癌细胞的现象，探讨Piezo1用于合成生物学肿瘤转移诊断基因线路的可行性

Synthetic biology utilizes engineering approach to adapt, improve and design biological components, construct synthetic gene circuits. These circuits can be used to optimize cellular programs for information processes and other functions according to human design. Synthetic biology is in the process of helping fight against diseases and improving human health. Mechanical sensing is one of the most fundamental and critical biological signal processes. However, there is no successful attempt reported for engineering mechanical sensing for synthetic biology applications. Here we perform systematic quantitative characterization of Piezo1, the most promising mechanical sensing ion channel. Specifically, we investigate how the interactions between it and the host cell affect its sensing to static stress and sonic wave. By using a synthetic expression tool to reduce heterogeneity in exogenous gene expression, we observed two novel phenomena, Piezo1 channel opening is response to shear stress in a biphasical manner, and it is sensitive to a specific frequency of sound vibrations. Before go along with engineering Piezo1 for synthetic biology applications, we will investigate the possibility of desensitization by larger shear stress, and if the cytoskeleton of host cell serves as a sound vibration filter. Towards these efforts, we will a technology to enable direct apply high frequency vibration to membrane or cytoskeleton of host cell. Finally we will express Piezo1 into A549 cells, to determine if it could sense the stiffness increase accompany EMT and metastasis, and investigate the viability of using it to construct synthetic circuit to detect cancer metastasis.