Background:

Estrogen receptor (ESR1) mutations occur at a high frequency in metastatic breast tumors in patients treated with hormonal therapy in the metastatic setting. We do not know if these mutations changed metabolomics and whether these metabolomics change could affect metastasis.

Experimental design and methods:

We generated ESR1 Y537S homozygous mutations using CRISPR Casp-9 technology. Globe metabolites screening was performed using 6550 Agilent QTOF instrument. Athymic mice were used in tumor xenograft studies. Affymetrix microarrays were performed to compare gene expression changes in Y537S mutant compared with parental cells. Enriched metabolite pathways and gene expression integrated analysis was analyzed by using online analysis tool http://www.metaboanalyst.ca.

Results:

We generated CRISPR ESR1 Y537S mutation homozygous knock-in clones in MCF-7 cells. In vivo experiments revealed that mutant cells are dominant drivers of metastasis. Transcriptome profiling revealed elevated expression of Hallmark pathways, including EMT and estrogen-regulated gene expression. We performed globe metabolites screening using MCF-7 Y537S and MCF-7 parental and identified 134 metabolites. Serum starvation media was used and estrogen was used as control for both cell lines. As we expected estrogen treatment induced metabolites changes in parental cells. However, metabolites in mutant cells were not changed significantly under estrogen treatment. Interestingly, metabolites in the mutant cells at baseline were remarkably upregulated (78 out of 134 identified total metabolites) indicating mutant cells in serum starvation condition had significantly different metabolomics compared with parental cells. Top upregulated pathways include protein biosynthesis, betaine metabolism and ammonia recycling. Integration of microarray gene expression and metabolites reviewed several metabolomics pathways significantly changed in mutant compared with parental cells including for example aminoacyl-tRNA biosynthesis, arginine and proline metabolism and alanine, aspartate and glutamate metabolism.

Conclusion: The Y537S ER mutation is a driver of distant metastasis in ER-positive breast cancer cells. Y537S ER mutant had globe changes of metabolites expression which was confirmed by integrated analysis combining microarray gene expression. The roles of these metabolites need to be studied to correlate with metastasis. Enzymes responsible for converting these metabolites changes could be served as potential therapeutic targets.

Citation Format: Gu G, Piyarathna B, Coarfa C, Ellis L, Ando' S, Fuqua S. The Y537S ESR1 mutation carries unique metabolomics profiling in breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P1-02-03.