Abstract 375: Lipogenic phenotype is an adaptation to nutrient stress in the tumor microenvironment

Abstract The nutrient and oxygen poor microenvironment combined with acidosis arising from fermentative metabolism imparts strong selection pressure on cancer cells that promotes evolution of more aggressive cancer types. Poor nutrient availability in the tumor microenvironment forces cancer cells to adapt to alternate routes of metabolism, nutrient scavenging and autophagy leading to a shift in their metabolic phenotype. These adaptations promote evolution of invasive and aggressive phenotypes. To test this hypothesis and to characterize the evolution of cancer cells under nutrient stress, we generated nutrient stress adapted cells from MCF7-GFP cells by subjecting them to multiple cycles of nutrient starvation and repletion. Distinct clones that outgrew were selected as nutrient stress adapted cells (NSA). Here we show that MCF7-NSA cells that are adapted to grow in low nutrient conditions exhibited robust accumulation of lipid droplets compared to parental MCF7-GFP cells as shown by confocal microscopy after neutral lipid staining as well as immunofluorescence staining for PLIN2 (Perilipin 2), a lipid droplet coat protein. In addition, MCF7-NSA cells showed elevated expression of the nuclear receptor PPARγ and the adipokine FABP4 indicating an adipogenic/lipogenic signature. Strikingly, these nutrient stress adapted cells were capable of nutrient scavenging by macropinocytosis compared to control cells even under nutrient replete conditions. Macropinocytosis was assessed by DQ-BSA processing and TMR-Dextran uptake. NHE inhibitor EIPA (ethyl isopropyl amiloride), that blocks macropinocytosis, but not receptor mediated endocytosis confirmed dextran uptake through macropinocytosis. Similarly, macropinocytosis inhibitor KH7 blocked dextran uptake in these adapted cells. Furthermore, MCF7-NSA cells showed high expression of TFEB and TFE3, master regulators of autophagy and lysosomogenesis suggesting that the necrotic cell debris scavenged through macropinocytosis and processed through autolysosome machinery might be contributing to de novo lipogenesis. Although control and adapted cells exhibited similar growth kinetics in vitro, when implanted in mammary fat pads (MFP) of NSG mice, NSA cells formed significantly smaller and slow growing primary tumors. Interestingly, 80% of these mice showed metastasis to other organs (lung, liver, kidney) whereas control mice did not develop metastasis as parental MCF7 is not an aggressive cell line in xenograft models. MCF7-NSA cells were more migratory in vitro in wound healing assays and were capable of developing lung metastases in a mouse model where primary tumor from mammary fat pad was resected to allow for metastases to develop. Taken together, our data suggest that lipogenic phenotype confers survival advantage in the harsh tumor microenvironment and selects for more invasive and aggressive phenotypes. Citation Format: Crystal Griffith, Dominique Abrahams, Antonio Ortiz, Alexandra Tassielli, Joseph Johnson, Robert Gatenby, Smitha Pillai. Lipogenic phenotype is an adaptation to nutrient stress in the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 375.