Researchers use mitochondrial uncouplers to target breast cancer metastasis

Mitochondrial serine catabolic pathway (MSCP), also called serine, glycine, and one-carbon metabolism, is essential for cancer cells to produce amino acids, one-carbon units, and reducing equivalents. The pathway is frequently upregulated in tumors: serine hydroxymethyltransferase 2 (SHMT2) and methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) are among the most overexpressed metabolic enzymes in human cancers, and their activity supports cancer cell proliferation, redox balance, and metastasis. Cancer cells rely on MSCP when the electron transport chain (ETC) is impaired, yet no effective therapy selectively targets this pathway. Fat mass and obesity-associated protein (FTO), an m⁶A demethylase, can be inhibited by 2-hydroxyglutarate (2-HG). FTO expression is associated with favorable prognosis and is downregulated in breast cancer, yet there is currently no effective way to reactivate FTO in cancer.Given these challenges, there is a need for in-depth research on therapies that safely reactivate respiration and target MSCP in metastatic breast cancer.

Researchers at Stanford University School of Medicine, with collaborators at The Second Hospital of Jilin University and Agilent Technologies, published (DOI: 10.20892/j.issn.2095-3941.2025.0444) the study in Cancer Biology & Medicine. The team, led by Jiangbin Ye, used uniformly ¹³C-labeled serine (U-¹³C-serine) tracing, immunoblotting, methylated RNA immunoprecipitation quantitative polymerase chain reaction (MeRIP-qPCR), and mouse metastasis models to test whether mitochondrial uncouplers such as niclosamide ethanolamine (NEN) and BAM15 suppress MSCP activity and breast cancer metastasis.

Using U-¹³C-serine tracing, the researchers found that NEN and BAM15 did not change serine uptake but sharply reduced glycine and purine nucleotides labeling, indicating lower MSCP and de novo purine synthesis flux. Uncouplers increased oxygen consumption and the NAD⁺ (nicotinamide adenine dinucleotide): NADH (reduced nicotinamide adenine dinucleotide) ratio, confirming ETC activation and alleviated reductive stress. Prolonged treatment downregulated mitochondrial enzymes SHMT2, MTHFD2, and methylenetetrahydrofolate dehydrogenase 1-like (MTHFD1L) proteins in triple-negative breast cancer (TNBC) and pancreatic cancer cells, while cytosolic enzyme serine hydroxymethyltransferase 1 (SHMT1) protein remained unaffected.

Surprisingly, mRNA levels of SHMT2, MTHFD2, and MTHFD1L were unchanged or elevated, pointing to post-transcriptional control. Both pharmacologic or genetic inhibition of FTO restored MSCP enzyme expression. Uncoupler treatment reduced 2-HG and global m⁶A levels; L-2-HG supplementation increased m⁶A and MSCP enzymes, whereas α-KG (alpha-ketoglutarate) supplementation decreased them. In mice, a 2000 ppm NEN diet significantly reduced breast cancer lung metastasis. Low FTO expression was associated with poorer relapse-free survival in breast cancer patients, while SHMT2 was overexpressed in metastatic samples. Given that FTO is an α-KG-dependent dioxygenase and 2-HG acts as a competitive inhibitor, these findings suggest that, by lowering 2-HG, uncoupling restores FTO activity and reduces m⁶A marks on MSCP transcripts.

The authors said the findings reframe the Warburg effect as a targetable metabolic state. They said reactivating the electron transport chain (ETC) with mitochondrial uncouplers restores the NAD⁺:NADH balance, lowers 2-HG, and releases FTO to demethylate m⁶A on MSCP transcripts. In their view, this makes mitochondrial uncoupling a promising way to attack metabolic vulnerabilities that metastatic breast cancer cells rely on. They emphasized that the mechanism links metabolism, epitranscriptome, and metastasis in one axis.

The study points to mitochondrial uncouplers as potential therapeutics for metastatic breast cancer, especially tumors with high SHMT2 expression. Niclosamide, the parent of NEN, is approved as an antiparasitic drug but has shown limited oncology efficacy, possibly due to poor bioavailability or a narrow therapeutic window. BAM15 and next-generation uncouplers with broader dosing windows may be better candidates. Future work should test these agents in clinical trials, and determine whether combining uncoupling with existing therapies can delay or prevent metastasis. Combining metabolic therapy with standard treatments may further improve outcomes and help turn this metabolic strategy into a practical treatment option.