ADHFE1 is a breast cancer oncogene and induces metabolic reprogramming
Prachi Mishra, … , Nagireddy Putluri, Stefan Ambs
Prachi Mishra, … , Nagireddy Putluri, Stefan Ambs
Published November 27, 2017
Citation Information: J Clin Invest. 2018;128(1):323-340. https://doi.org/10.1172/JCI93815.
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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 1

Co-occurrence of ADHFE1 and MYC amplifications in human breast tumors and accelerated tumor growth of ADHFE1-overexpressing MCF7 cells.

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Co-occurrence of ADHFE1 and MYC amplifications in human breast tumors an...
(A) Accumulation of D- and L-2-hydroxyglutarate and their ratio (D to L ratio above bars) in 15 human breast tumors and 4 adjacent noncancerous breast tissues (N). (B) Co-occurrence of ADHFE1 and MYC amplifications in human breast tumors (P < 0.001). Shown are 473 TCGA breast tumors with amplification (dark red) or overexpression (light red) of 1 of the 5 listed genes previously associated with D-2HG. Blue bars: deletion or reduced expression. IDH, isocitrate dehydrogenase; PHGDH, phosphoglycerate dehydrogenase. (C) High ADHFE1 protein expression by immunohistochemistry (IHC) is associated with decreased survival of patients with ER-negative breast cancer. P = 0.012 by the log-rank test. HR, hazard ratio. (D) Western blots. Top left panel: MYC upregulates ADHFE1. MYC signaling was induced (MYC-ER fusion in HMEC-MYC) or suppressed (inducible shRNA in SUM159T) with 4-hydroxytamoxifen (+) or doxycycline (+), respectively. HMEC, human mammary epithelial cell. Lower panel: ADHFE1 transgene expression in MCF10A and MCF12A cells increases MYC. Right panel: Induction of aldo-keto reductase AKR7A2, but not AKR1A1, in HMEC-MYC cells after 4-hydroxytamoxifen–stimulated MYC signaling (+TAM). (E) Increased tumor growth of MCF7 cells over-expressing either ADHFE1, MYC, or both ADHFE1 and MYC (ADHFE1-MYC). Solid lines show median for each group. Tumor growth was measured 16 weeks after injection of MCF7 cells into mammary fat pads. Two-sided t test for comparisons with control group (n = 10 per group). P = 0.05 for MYC versus ADHFE1-MYC tumors. (F) Increased 2-hydroxyglutarate (2HG) and 4-hydroxybutyrate (4HB) levels in MCF7 tumors overexpressing ADHFE1 and MYC (n = 10 per group). *P < 0.05, **P < 0.01, versus control group using 2-sided t test. Shown is the mean ± SD. ANOVA test for differences between groups was used in E and F: P < 0.001.