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荷黑素瘤小鼠中肿瘤细胞和免疫细胞的单细胞代谢成像

Single cell metabolic imaging of tumor and immune cells in melanoma bearing mice.

作者信息

Heaton Alexa R, Rehani Peter R, Hoefges Anna, Lopez Angelica F, Erbe Amy K, Sondel Paul M, Skala Melissa C

机构信息

Morgridge Institute for Research, Madison, WI, United States.

Department of Human Oncology, University of Wisconsin, Madison, WI, United States.

出版信息

Front Oncol. 2023 Mar 20;13:1110503. doi: 10.3389/fonc.2023.1110503. eCollection 2023.

DOI:10.3389/fonc.2023.1110503
PMID:37020875
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10067577/
Abstract

INTRODUCTION

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 images of cell metabolism at a single cell level.

METHODS

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 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 images.

RESULTS

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 .

DISCUSSION

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 context.

摘要

引言

癌症和免疫细胞的代谢重编程发生在肿瘤发生过程中，对癌症进展有重大影响。不幸的是，目前测量肿瘤和免疫细胞代谢的技术需要样本破坏和/或细胞分离，这会消除空间背景。对自发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）进行双光子荧光寿命成像显微镜（FLIM）可在单细胞水平提供细胞代谢图像。

方法

在此，我们报告一种免疫活性mCherry报告小鼠模型，用于在分化过程中表达CD4或在成熟状态下表达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成像。我们还报告了一种算法，用于在图像中对表达mCherry的免疫细胞进行单细胞分割。

结果

我们发现，与脾脏中的免疫细胞相比，B78肿瘤内的免疫细胞FAD平均寿命降低，结合型FAD比例增加。与脾脏中的免疫细胞相比，肿瘤浸润免疫细胞的大小也增加。这些变化与肿瘤浸润免疫细胞相比脾脏免疫细胞向激活和增殖增加的转变一致。与同一肿瘤内的B78肿瘤细胞相比，肿瘤浸润免疫细胞的FAD平均寿命增加，蛋白质结合型FAD寿命增加。在免疫细胞和肿瘤细胞中均观察到单细胞代谢异质性。

讨论

这种方法可用于监测肿瘤细胞和免疫细胞中的单细胞代谢异质性，以研究在自然环境中治疗癌症的有前景的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/593ca2b7ee03/fonc-13-1110503-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/7165b514c5c0/fonc-13-1110503-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/1ca52574c657/fonc-13-1110503-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/3581f3a0154d/fonc-13-1110503-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/3b5da5ca1b6b/fonc-13-1110503-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/9d185856191d/fonc-13-1110503-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/593ca2b7ee03/fonc-13-1110503-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/7165b514c5c0/fonc-13-1110503-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/1ca52574c657/fonc-13-1110503-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/3581f3a0154d/fonc-13-1110503-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/3b5da5ca1b6b/fonc-13-1110503-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/9d185856191d/fonc-13-1110503-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33fa/10067577/593ca2b7ee03/fonc-13-1110503-g006.jpg

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