ADHFE1 is a breast cancer oncogene and induces metabolic reprogramming
Prachi Mishra, Wei Tang, Vasanta Putluri, Tiffany H. Dorsey, Feng Jin, Fang Wang, Donewei Zhu, Lauren Amable, Tao Deng, Shaofei Zhang, J. Keith Killian, Yonghong Wang, Tsion Z. Minas, Harry G. Yfantis, Dong H. Lee, Arun Sreekumar, Michael Bustin, Wei Liu, Nagireddy Putluri, Stefan Ambs
Prachi Mishra, Wei Tang, Vasanta Putluri, Tiffany H. Dorsey, Feng Jin, Fang Wang, Donewei Zhu, Lauren Amable, Tao Deng, Shaofei Zhang, J. Keith Killian, Yonghong Wang, Tsion Z. Minas, Harry G. Yfantis, Dong H. Lee, Arun Sreekumar, Michael Bustin, Wei Liu, Nagireddy Putluri, Stefan Ambs
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Research Article Metabolism Oncology

ADHFE1 is a breast cancer oncogene and induces metabolic reprogramming

Abstract

Metabolic reprogramming in breast tumors is linked to increases in putative oncogenic metabolites that may contribute to malignant transformation. We previously showed that accumulation of the oncometabolite, 2-hydroxyglutarate (2HG), in breast tumors was associated with MYC signaling, but not with isocitrate dehydrogenase (IDH) mutations, suggesting a distinct mechanism for increased 2HG in breast cancer. Here, we determined that D-2HG is the predominant enantiomer in human breast tumors and show that the D-2HG–producing mitochondrial enzyme, alcohol dehydrogenase, iron-containing protein 1 (ADHFE1), is a breast cancer oncogene that decreases patient survival. We found that MYC upregulates ADHFE1 through changes in iron metabolism while coexpression of both ADHFE1 and MYC strongly enhanced orthotopic tumor growth in MCF7 cells. Moreover, ADHFE1 promoted metabolic reprogramming with increased formation of D-2HG and reactive oxygen, a reductive glutamine metabolism, and modifications of the epigenetic landscape, leading to cellular dedifferentiation, enhanced mesenchymal transition, and phenocopying alterations that occur with high D-2HG levels in cancer cells with IDH mutations. Together, our data support the hypothesis that ADHFE1 and MYC signaling contribute to D-2HG accumulation in breast tumors and show that D-2HG is an oncogenic metabolite and potential driver of disease progression.

Authors

Prachi Mishra, Wei Tang, Vasanta Putluri, Tiffany H. Dorsey, Feng Jin, Fang Wang, Donewei Zhu, Lauren Amable, Tao Deng, Shaofei Zhang, J. Keith Killian, Yonghong Wang, Tsion Z. Minas, Harry G. Yfantis, Dong H. Lee, Arun Sreekumar, Michael Bustin, Wei Liu, Nagireddy Putluri, Stefan Ambs

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Figure 2

ADHFE1 promoter activity in HMEC-MYC cells and induction of ADHFE1 protein expression by FeSO4.

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ADHFE1 promoter activity in HMEC-MYC cells and induction of ADHFE1 prot...
(A) HMEC-MYC cells were transfected with a reporter construct containing a 5-kb segment of the human ADHFE1 promoter region with a MYC binding site. Cells were transfected with constructs containing either the intact MYC binding site (WT) or a mutated sequence (mutant), as described in Methods. MYC signaling was induced with 4-hydroxytamoxifen (+TAM). Addition of TAM modestly increased ADHFE1 promoter activity but the activity was not dependent on the presence of an intact MYC binding site. Shown is the mean ± SD for triplicate experiments. Two-sided t test; NS, not significantly different. (B) MYC signaling (+TAM) increases expression of iron metabolism genes in HMEC-MYC cells. (C) MYC signaling also increases mitochondrial iron content in these cells. n = 4. Shown is the mean ± SD. *P < 0.05 (2-sided t test). (D) Treatment of HMECs with FeSO4 for 24 hours increases ADHFE1 expression. (E) Induction of ADHFE1 and iron metabolism genes by MYC signaling in HMEC-MYC cells can be inhibited by the Fe2+ chelator, 3-AP (0.5 μM), which was added together with TAM. In B, C, and E, TAM treatment was for 96 hours. TFRC, transferrin receptor protein 1; IRP2, iron-responsive element–binding protein 2.