Single cell metabolic imaging of tumor and immune cells in vivo in melanoma bearing mice.
Single cell metabolic imaging of tumor and immune cells in vivo in melanoma bearing mice.
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DOI：
10.3389/fonc.2023.1110503
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发表时间：
2023
影响因子：
4.7
通讯作者：
Skala, Melissa C.
中科院分区：
文献类型：
--
作者：
Heaton, Alexa R.;Rehani, Peter R.;Hoefges, Anna;Lopez, Angelica F.;Erbe, Amy K.;Sondel, Paul M.;Skala, Melissa C.
关键词：
癌症和免疫细胞的代谢重编程发生在肿瘤发生过程中，对癌症的进展有重要影响。不幸的是，目前测量肿瘤和免疫细胞代谢的技术需要样品破坏和/或细胞分离，从而消除空间背景。自荧光代谢辅酶烟酰胺腺嘌呤二核苷酸（磷酸）（NAD(P)H）和黄素腺嘌呤二核苷酸（FAD）的双光子荧光寿命成像显微镜（FLIM）提供了单细胞水平的细胞代谢的体内图像。在这里，我们报告了一个免疫活性的mCherry报告小鼠模型，用于在分化过程中表达CD4或成熟状态下表达CD4和/或CD8的免疫细胞，并在同基因B78黑色素瘤模型中对免疫细胞和癌细胞进行体内成像。我们还报道了一种在体内图像中表达mccherry的免疫细胞的单细胞分割算法。我们发现，与脾脏内的免疫细胞相比，B78肿瘤内的免疫细胞的FAD平均寿命缩短，结合FAD的比例增加。肿瘤浸润免疫细胞的大小也比脾脏免疫细胞增大。这些变化与肿瘤浸润免疫细胞与脾脏免疫细胞相比，向激活和增殖增加的转变是一致的。与同一肿瘤内的B78肿瘤细胞相比，肿瘤浸润免疫细胞的FAD平均寿命和蛋白结合FAD寿命均增加。在体内免疫细胞和肿瘤细胞中均观察到单细胞代谢异质性。这种方法可用于监测肿瘤细胞和免疫细胞的单细胞代谢异质性，从而在体内环境中研究有希望的癌症治疗方法。
Metabolic reprogramming of cancer and immune cells occurs during tumorigenesis and has a significant impact on cancer progression. Unfortunately, current techniques to measure tumor and immune cell metabolism require sample destruction and/or cell isolations that remove the spatial context. Two-photon fluorescence lifetime imaging microscopy (FLIM) of the autofluorescent metabolic coenzymes nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD) provides in vivo images of cell metabolism at a single cell level. Here, we report an immunocompetent mCherry reporter mouse model for immune cells that express CD4 either during differentiation or CD4 and/or CD8 in their mature state and perform in vivo imaging of immune and cancer cells within a syngeneic B78 melanoma model. We also report an algorithm for single cell segmentation of mCherry-expressing immune cells within in vivo images. We found that immune cells within B78 tumors exhibited decreased FAD mean lifetime and an increased proportion of bound FAD compared to immune cells within spleens. Tumor infiltrating immune cell size also increased compared to immune cells from spleens. These changes are consistent with a shift towards increased activation and proliferation in tumor infiltrating immune cells compared to immune cells from spleens. Tumor infiltrating immune cells exhibited increased FAD mean lifetime and increased protein-bound FAD lifetime compared to B78 tumor cells within the same tumor. Single cell metabolic heterogeneity was observed in both immune and tumor cells in vivo. This approach can be used to monitor single cell metabolic heterogeneity in tumor cells and immune cells to study promising treatments for cancer in the native in vivo context.
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影响因子：
5.8
作者：
Gu, Zuguang;Eils, Roland;Schlesner, Matthias
通讯作者：
Schlesner, Matthias
影响因子：
10.9
作者：
Aiken TJ;Komjathy D;Rodriguez M;Stuckwisch A;Feils A;Subbotin V;Birstler J;Gillies SD;Rakhmilevich AL;Erbe AK;Sondel PM
通讯作者：
Sondel PM
影响因子：
5.4
作者：
Cham, Candace M.;Driessens, Gregory;O'Keefe, James P.;Gajewski, Thomas F.
通讯作者：
Gajewski, Thomas F.
影响因子：
48
作者：
Cutler, Kevin J.;Stringer, Carsen;Lo, Teresa W.;Rappez, Luca;Stroustrup, Nicholas;Peterson, S. Brook;Wiggins, Paul A.;Mougous, Joseph D.
通讯作者：
Mougous, Joseph D.
影响因子：
9.8
作者：
Haas R;Smith J;Rocher-Ros V;Nadkarni S;Montero-Melendez T;D'Acquisto F;Bland EJ;Bombardieri M;Pitzalis C;Perretti M;Marelli-Berg FM;Mauro C
通讯作者：
Mauro C