Asparagine synthetase and G‐protein coupled estrogen receptor are critical responders to nutrient supply in KRAS mutant colorectal cancer

• Published in: International journal of cancer 2025-01, Vol.156 (1), p.52-68
• Main Authors: Lu, Lingeng, Zhang, Qian, Aladelokun, Oladimeji, Berardi, Domenica, Shen, Xinyi, Marin, Audrey, Garcia‐Milian, Rolando, Roper, Jatin, Khan, Sajid A., Johnson, Caroline H.
• Format: Article
• Language: English
• Subjects: 17β-Estradiol; ASNS; Asparagine; Aspartate-ammonia ligase; Aspartate-Ammonia Ligase - genetics; Aspartate-Ammonia Ligase - metabolism; Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; Caspase; Cell growth; Cell Line, Tumor; Cell Proliferation; Cell survival; Clinical outcomes; Cohort analysis; Colon cancer; Colorectal cancer; Colorectal carcinoma; Colorectal Neoplasms - genetics; Colorectal Neoplasms - metabolism; Colorectal Neoplasms - pathology; estradiol; Estradiol - metabolism; Estradiol - pharmacology; Estrogen receptors; Estrogens; Female; Gene Expression Regulation, Neoplastic; Glutamine; Glutamine - metabolism; GPER1; Humans; K-Ras protein; Male; Molecular modelling; Mutants; Mutation; Nutrients - metabolism; Proto-Oncogene Proteins p21(ras) - genetics; Proto-Oncogene Proteins p21(ras) - metabolism; Receptors, Estrogen - metabolism; Receptors, G-Protein-Coupled - genetics; Receptors, G-Protein-Coupled - metabolism; Sex differences; survival; Tumors
• ISSN: 0020-7136; 1097-0215; 1097-0215
• DOI: 10.1002/ijc.35104

Survival differences exist in colorectal cancer (CRC) patients by sex and disease stage. However, the potential molecular mechanism(s) are not well understood. Here we show that asparagine synthetase (ASNS) and G protein‐coupled estrogen receptor‐1 (GPER1) are critical sensors of nutrient depletion and linked to poorer outcomes for females with CRC. Using a 3D spheroid model of isogenic SW48 KRAS wild‐type (WT) and G12A mutant (MT) cells grown under a restricted nutrient supply, we found that glutamine depletion inhibited cell growth in both cell lines, whereas ASNS and GPER1 expression were upregulated in KRAS MT versus WT. Estradiol decreased growth in KRAS WT but had no effect on MT cells. Selective GPER1 and ASNS inhibitors suppressed cell proliferation with increased caspase‐3 activity of MT cells under glutamine depletion condition particularly in the presence of estradiol. In a clinical colon cancer cohort from The Cancer Genome Atlas, both high GPER1 and ASNS expression were associated with poorer overall survival for females only in advanced stage tumors. These results suggest KRAS MT cells have mechanisms in place that respond to decreased nutrient supply, typically observed in advanced tumors, by increasing the expression of ASNS and GPER1 to drive cell growth. Furthermore, KRAS MT cells are resistant to the protective effects of estradiol under nutrient deplete conditions. The findings indicate that GPER1 and ASNS expression, along with the interaction between nutrient supply and KRAS mutations shed additional light on the mechanisms underlying sex differences in metabolism and growth in CRC, and have clinical implications in the precision management of KRAS mutant CRC. What's new? The mechanisms underlying survival differences in colorectal cancer by sex and disease stage remain unclear. Here, in a 3D spheroid model under restricted nutrient supply (as in advanced cancers), estradiol was protective of colorectal cancer growth in KRAS wild‐type but not mutant cells, which had increased GPER1 and ASNS expression. In a clinical colorectal cancer dataset, high GPER1 and ASNS expression was associated with poorer prognosis in females with advanced colorectal cancer, but not in males. These findings suggest that ASNS and GPER1 are responsive to the nutrient microenvironment and can restore colorectal cancer cell growth in females.