肿瘤代谢的异质性是肿瘤生物学的一个重要但仍知之甚少的方面。目前的工作重点是使用氧化还原辅因子 NAD(P)H 的荧光寿命成像 (FLIM) 对结直肠癌细胞代谢异质性进行可视化和量化。在体外四种癌细胞系（HT29、HCT116、CaCo2 和 CT26）、体内四种类型的小鼠结直肠肿瘤以及离体患者肿瘤样本中对 NAD(P)H 进行了 FLIM 显微镜检查。评估衰减参数的分散性和双峰性以量化细胞间代谢异质性。我们的结果表明，与培养细胞和肿瘤异种移植物相比，患者的结直肠肿瘤具有显着更高的能量代谢异质性，这表现为内部 NAD(P)H 游离（糖酵解）部分的贡献更广泛且频繁的双峰分布。一个样本。在患者的肿瘤中，高级别和早期肿瘤的离散度较大，但与双峰性没有任何关联。这些结果表明，通过 NAD(P)H FLIM 评估的细胞水平代谢异质性有可能成为临床预后因素。© 2024，Komarova、Sinyushkina、Shchechkin 等人。

Heterogeneity of tumor metabolism is an important, but still poorly understood aspect of tumor biology. Present work is focused on the visualization and quantification of cellular metabolic heterogeneity of colorectal cancer using fluorescence lifetime imaging (FLIM) of redox cofactor NAD(P)H. FLIM-microscopy of NAD(P)H was performed in vitro in four cancer cell lines (HT29, HCT116, CaCo2 and CT26), in vivo in the four types of colorectal tumors in mice and ex vivo in patients' tumor samples. The dispersion and bimodality of the decay parameters were evaluated to quantify the intercellular metabolic heterogeneity. Our results demonstrate that patients' colorectal tumors have significantly higher heterogeneity of energy metabolism compared with cultured cells and tumor xenografts, which was displayed as a wider and frequently bimodal distribution of a contribution of a free (glycolytic) fraction of NAD(P)H within a sample. Among patients' tumors, the dispersion was larger in the high-grade and early stage ones, without, however, any association with bimodality. These results indicate that cell-level metabolic heterogeneity assessed from NAD(P)H FLIM has a potential to become a clinical prognostic factor.© 2024, Komarova, Sinyushkina, Shchechkin et al.