Nashimoto Yuji, Shishido Shotaro, Onuma Kunishige, Ino Kosuke, Inoue Masahiro, Shiku Hitoshi
Institute of Bioengineering and Biomaterials (IBB), Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
Frontier Research Institute for Interdisciplinary Sciences (FRIS), Tohoku University, Sendai, Miyagi, Japan.
Front Bioeng Biotechnol. 2023 May 18;11:1184325. doi: 10.3389/fbioe.2023.1184325. eCollection 2023.
Heterogeneous nature is a pivotal aspect of cancer, rendering treatment problematic and frequently resulting in recurrence. Therefore, advanced techniques for identifying subpopulations of a tumour in an intact state are essential to develop novel screening platforms that can reveal differences in treatment response among subpopulations. Herein, we conducted a non-invasive analysis of oxygen metabolism on multiple subpopulations of patient-derived organoids, examining its potential utility for non-destructive identification of subpopulations. We utilised scanning electrochemical microscopy (SECM) for non-invasive analysis of oxygen metabolism. As models of tumours with heterogeneous subpopulations, we used patient-derived cancer organoids with a distinct growth potential established using the cancer tissue-originated spheroid methodology. Scanning electrochemical microscopy measurements enabled the analysis of the oxygen consumption rate (OCR) for individual organoids as small as 100 µm in diameter and could detect the heterogeneity amongst studied subpopulations, which was not observed in conventional colorectal cancer cell lines. Furthermore, our oxygen metabolism analysis of pre-isolated subpopulations with a slow growth potential revealed that oxygen consumption rate may reflect differences in the growth rate of organoids. Although the proposed technique currently lacks single-cell level sensitivity, the variability of oxygen metabolism across tumour subpopulations is expected to serve as an important indicator for the discrimination of tumour subpopulations and construction of novel drug screening platforms in the future.
异质性是癌症的一个关键特征，这使得治疗变得困难，并常常导致复发。因此，用于完整状态下识别肿瘤亚群的先进技术对于开发能够揭示亚群间治疗反应差异的新型筛查平台至关重要。在此，我们对患者来源的类器官的多个亚群进行了氧代谢的非侵入性分析，研究其在非破坏性识别亚群方面的潜在效用。我们利用扫描电化学显微镜（SECM）对氧代谢进行非侵入性分析。作为具有异质性亚群的肿瘤模型，我们使用了通过癌症组织来源的球体方法建立的具有不同生长潜能的患者来源的癌症类器官。扫描电化学显微镜测量能够分析直径小至100 µm的单个类器官的氧消耗率（OCR），并能检测所研究亚群之间的异质性，这在传统的结肠癌细胞系中未观察到。此外，我们对预分离的具有缓慢生长潜能的亚群进行的氧代谢分析表明，氧消耗率可能反映类器官生长速率的差异。尽管所提出的技术目前缺乏单细胞水平的灵敏度，但肿瘤亚群间氧代谢的变异性有望在未来作为区分肿瘤亚群和构建新型药物筛选平台的重要指标。
