Development of next-generation ion conductance microscopy to unravel the phenotype and gene expression state of cancer cells

开发下一代离子电导显微镜以揭示癌细胞的表型和基因表达状态

基本信息

批准号：
21H01770
负责人：
Korchev Yuri.E.
金额：
$ 11.15万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2021
资助国家：
日本
起止时间：
2021-04-01 至 2025-03-31
项目状态：
未结题

项目摘要

1. We have established a new SICM quantitative nanoscale mapping technique to measure the stiffness of breast cancer MCF7 cells and skin cancer melanoma A375M cells without damaging the cells. In addition, we formed a Pt-functionalized carbon nanoelectrode at the tip of nanopipette to measure the ROS distribution of individual cancer cells. Now we can phenotype individual breast cancer MCF7 cells and skin cancer melanoma A375M cells with their stiffness and ROS distribution at single cell level.2. It is becoming clear that an acid pHe plays an essential role in cancer cell progression, invasiveness and resistance to therapy. Monitoring the pHe variation of cancer cells will provide critical information for understanding tumour heterogeneity during pathological processes such as epithelial-mesenchyme transition. Based on our previously studies on enzyme-based pH-sensing nanoprobes, we have developed non-enzyme pH-sensing nanoprobes for pHe measuring from individual cancer cells.3. Regarding gene expression evaluation, conventional methods disrupt cells and cause the instability in their results. It has been difficult to know the intracellular gene expression status of individual cancer cells. Gene sampling with SICM nanopipette can perform repeat sampling from the same cell at different time, which allows a dynamical gene expression evaluation. Under the help of SICM biopsy sampling methods, we have evaluated the mRNA expression in cytoplasm of individual cancer cells.

1.我们建立了一种新的SICM定量纳米级绘图技术，可以在不损伤细胞的情况下测量乳腺癌MCF7细胞和皮肤癌黑色素瘤A375M细胞的硬度。此外，我们在纳米移液器尖端形成了铂功能化碳纳米电极，以测量单个癌细胞的ROS分布。现在我们可以在单细胞水平上对单个乳腺癌MCF7细胞和皮肤癌黑色素瘤A375M细胞进行表型分析，了解它们的硬度和ROS分布。2．越来越清楚的是，酸性 pHe 在癌细胞进展、侵袭性和治疗耐药性中发挥着重要作用。监测癌细胞的 pHe 变化将为了解上皮-间质转化等病理过程中肿瘤异质性提供关键信息。基于我们之前对基于酶的pH传感纳米探针的研究，我们开发了非酶pH传感纳米探针，用于测量单个癌细胞的pHe。 3．在基因表达评估方面，传统方法会破坏细胞并导致结果不稳定。了解单个癌细胞的细胞内基因表达状态一直很困难。使用 SICM 纳米移液器进行基因采样可以在不同时间对同一细胞进行重复采样，从而实现动态基因表达评估。在SICM活检取样方法的帮助下，我们评估了单个癌细胞细胞质中的mRNA表达。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Phycosphere pH of unicellular nano- and micro- phytoplankton cells and consequences for iron speciation.

DOI：
10.1038/s41396-022-01280-1
发表时间：
2022-10
期刊：
ISME JOURNAL
影响因子：
11
作者：
Liu, Fengjie;Gledhill, Martha;Tan, Qiao-Guo;Zhu, Kechen;Zhang, Qiong;Salaun, Pascal;Tagliabue, Alessandro;Zhang, Yanjun;Weiss, Dominik;Achterberg, Eric P.;Korchev, Yuri
通讯作者：
Korchev, Yuri