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. 2021:2265:363-376.
doi: 10.1007/978-1-0716-1205-7_26.

Fate Mapping of Cancer Cells in Metastatic Lymph Nodes Using Photoconvertible Proteins

Ethel R Pereira  1   2 , Dmitriy Kedrin  1   3 , Timothy P Padera  4
Affiliations

Fate Mapping of Cancer Cells in Metastatic Lymph Nodes Using Photoconvertible Proteins

Ethel R Pereira et al. Methods Mol Biol. 2021.

Abstract

The lymph node microenvironment is extremely dynamic and responds to immune stimuli in the host by reprogramming immune, stromal, and endothelial cells. In normal physiological conditions, the lymph node will initiate an appropriate immune response to clear external threats that the host may experience. However, in metastatic disease, cancer cells often colonize local lymph nodes, disrupt immune function, and even leave the lymph node to create additional metastases. Understanding how cancer cells enter, colonize, survive, proliferate, and interact with other cell types in the lymph node is challenging. Here, we describe the use of photoconvertible fluorescent proteins to label and trace the fate of cancer cells once they enter the lymph node.

Keywords: Circulating tumor cells; Confocal microscopy; Dendra2; Intravital imaging; Lymph node; Metastasis; Photoconvertible proteins; Photodiode.

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Figures

Fig. 1
Fig. 1
Fate-mapping cancer cells in the lymph node. To trace cancer cells from the lymph node to distant organs, we engineer murine cancer cell lines to express the photoconvertible protein Dendra2. In the absence of light conversion, the primary tumor fluoresces green. Spontaneous lymphatic metastasis from the primary tumor results in cancer cell colonization in the lymph node in the mouse models we tested, which include B16F10 (melanoma), 4T1 (breast cancer), and SCCVII (squamous cell carcinoma). Green fluorescently labeled cancer cells in the lymph node are photoconverted to red fluorescence using a 405 nm photodiode. The red fluorescently labeled cancer cells in the lymph node are followed over time to trace their path inside the lymph node and beyond to distant organs such as the lung and liver. This is a powerful technology that can be utilized to trace the movement of cancer cells from a specific location through the complex metastatic cascade to their eventual colonization of secondary sites
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