Presentation: 2025 ND EPSCoR Annual conference
October 21, 2025, NDSU Memorial Union, Fargo, North Dakota
How Does RAGE Communicate with Immune Cells in the Tumor Microenvironment?
Peyton
Zaun
Doctoral Student
North Dakota State University
Co-authors: Yousuf Alam, Doctoral Student, North Dakota State University, Md Zahidul Hasan, Doctoral Student, North Dakota State University, Hilde van Gijssel, Professor, Valley City State University, Estelle Leclerc, Associate Professor, North Dakota State University
Session
Poster number: 155
Nueta Room
The receptor for advanced glycation end-products (RAGE) promotes melanoma cell proliferation, migration, and invasion as well as tumor growth, chemoresistance, and metastasis. The goal of this study was to determine if RAGE, expressed by cancer cells, communicates with immune cells, which are an important component of the tumor microenvironment. To reach our goal, we use a spheroid/ immune cells co-culture model. Two cell lines deriving from B16F10 murine melanoma cells were used to form the spheroids: the parental B16F10 RAGE (+), and the B16F10 RAGE (-), which was engineered using the CRISPR/Cas9 gene editing tool and lacks RAGE expression. Spheroids were formed from the melanoma cells using 250-500 cells/well in media supplemented with 0.12% methylcellulose to promote cell aggregation. The size of the spheroids was measured daily and the diameter estimated using ImageJ analysis of brightfield microscopic images. The immune cells (including CD4+, CD8+, and Natural Killer cells) were obtained from the spleen of an immune competent C57BL6 mouse. We successfully generated spherical spheroids from B16F10 RAGE (+). However, we are currently optimizing the experimental conditions with the B16F10 (-) cells as these cells do not currently form spherical spheroids. Initial co-culture experiments with B16F10 RAGE (+) spheroids and murine splenocytes suggest that the presence of immune cells in the spheroids reduces the growth rate of the spheroids. We are currently studying if the presence of RAGE in the spheroids affect the population of immune cells in the co-culture by flow cytometry.