Development of a method for the analysis of single-cell and single-molecule enzyme activities from oral cancer liquid biopsy.

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K19637
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 57:Oral science and related fields
• Research Institution: Nippon Medical School
• Principal Investigator: Honda Kazufumi 日本医科大学, 大学院医学研究科, 大学院教授 (10260936)
• Co-Investigator(Kenkyū-buntansha): 吉本 世一 国立研究開発法人国立がん研究センター, 中央病院, 科長 (00462242) ; 本間 義崇 国立研究開発法人国立がん研究センター, 中央病院, 医長 (30719943)
• Project Period (FY): 2022-06-30 – 2024-03-31
• Keywords: 1細胞酵素活性 / 口腔がん

Outline of Final Research Achievements

We have established a technical theory for encapsulating cancer cells in microwells one by one. Oral cancer cell lines were encapsulated for each cell, and the enzyme activity of each cell was confirmed, and MMP activity was confirmed, and (1) each cell retained the property of continuously enhancing MMP activity in the culture medium, and (2) there were cells in which MMP activity could not be confirmed in the culture medium even after a long period of time. These results can be interpreted as secretory MMPs that control the invasion of cancer cells represented by secretory MMP2 and MMP9 in (1) and (2). In this study, we observed for the first time in the world that heterogeneity exists in each cell of MMP. Single-cell enzyme activity is a completely new concept that explains the tumor microenvironment at high resolution.

Free Research Field

分子腫瘍学

Academic Significance and Societal Importance of the Research Achievements

MMP活性に関してはzymographyなどにより細胞全体が持つ活性を計測できる従来技術が存在した。しかしながら、１細胞毎でその活性を計測する方法は確立されていない。MMPの存在はがん手術材料の免疫染色などを利用すれば その発現状況は確認できるが、1細胞活性状態を評価する技術解析法は全くない。今回の研究結果から、がん細胞が持つMMPの酵素活性は1細胞毎にheterogeneityが存在することを世界で初めて観察した。1細胞毎の酵素活性heterogeneityはがん細胞巣の微小環境を高分解能で説明する全く新しい概念であり、がん細胞生物学を語るうえで大きなブレークスルーになると期待できる。