Abstract 2736: Decreasing colorectal cancer extracellular pH dampens immune cell killing and is partially ameliorated by treatment with NHE1 inhibitor Cariporide
Authors: Huntington Kelsey E. , Maxwell Aaron W. , Louie Anna , Zhang Shengliang , Zhou Lanlan , El-Deiry Wafik S. 
Published: July 2021

Abstract
Tumor cells upregulate a plethora of proteins that are important for pH regulation, resulting in the acidification of the extracellular tumor microenvironment. Abnormal pH is known to dampen immune function, resulting in a worsened anti-tumor immune response. Understanding how alterations in pH modulate the interactions between immune cells and tumors cells could help elucidate opportunities for new therapeutic approaches. To determine how pH modification impacts immune cell-mediated killing of tumor cells, co-culture experiments were conducted with GFP+ SW480 colorectal cancer cells and either NK-92 natural killer cells or TALL-104 T cells at various pHs in a 48-well plate. Manipulation of pH occurred using the compounds lactic acid and sodium bicarbonate. We observed that both NK-92 cells and TALL-104 cells exhibited decreasing amounts of cell killing in response to decreasing pH. Next, to determine if therapeutic manipulation of pH via alteration of pH pump activity could overcome the negative changes in immune cell activity associated with pH, we added Cariporide, a selective NHE1 inhibitor, to the co-culture experiments. Treatment with Cariporide bolstered T cell killing in a dose-dependent manner, however the doses used did not impact natural killer cell-mediated killing. Given the importance of cell signaling on immune function, and because we saw potentially significant changes in immune killing with Cariporide, we then evaluated how changes in pH using Cariporide affected these signaling molecules. Colorectal cancer cells (HCT116, HT29, KM12C) were treated with Cariporide at 5 µm and 10 µm doses and cancer cell culture supernatants were collected and frozen down at -80 °C. Cytokine, chemokine, and growth factor levels were analyzed using a Luminex 200 multiplexing instrument. Overall cytokine levels showed a decreasing trend in response to increasing doses of Cariporide. There were notable decreases in the levels of IL-8/CXCL8, VEGF, CCL3/MIP-1 alpha, and M-CSF with increasing doses of Cariporide. All of these factors, as well as extracellular acidosis, are known to promote epithelial mesenchymal transition (EMT), thus, promoting metastatic dissemination. A therapeutic that targets factors that lead to EMT could help prevent metastasis. Moreover, we saw that pH impacts the function of immune cells and this is relevant in the context of a highly acidic TME. Therapeutically modifying the pH of the TME using Cariporide treatment may be a way to improve immune response and prevent EMT. These experiments can help inform future investigations into how the pH of the tumor microenvironment may be extrinsically modulated to improve anti-tumor immune response.
Citation Format: Kelsey E. Huntington, Aaron W. Maxwell, Anna Louie, Shengliang Zhang, Lanlan Zhou, Wafik S. El-Deiry. Decreasing colorectal cancer extracellular pH dampens immune cell killing and is partially ameliorated by treatment with NHE1 inhibitor Cariporide [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2736.