Differential regulation of FADS2 by EZH2 reveals a metabolic vulnerability in ovarian cancer treatment.

Background: Ovarian cancer (OC) is the most aggressive and lethal gynaecological cancer and is characterised by abnormal lipid metabolism. Enhancer of zeste homologue 2 (EZH2) is the catalytic subunit of polycomb repressive complex 2 (PRC2). It functions as a tumour promoter through both canonical H3K27me3 modification and noncanonical mechanisms. Although EZH2 has been developed as a potential anticancer target, the therapeutic effects of EZH2 inhibitors in solid tumours are still limited.

Methods: The levels of fatty acid and tricarboxylic acid (TCA) cycle-related metabolites were measured via high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). EZH2-knockout cells, ChIP‒qPCR, qPCR, and western blotting were used to investigate the regulatory effect of EZH2 on FADS2. The expression of FADS2 transcript variants and their correlations with prognosis in clinical OC tissues were analysed via the GEPIA 2 database. Various OC cell lines, patient-derived organoids and tumour-bearing models have been used to test the therapeutic effects of EZH2 and FADS2 inhibitors. 4D-DIA proteomics, the Seahorse assay and flow cytometry-based detection of mitochondrial function and protein synthesis were performed to explore the mechanism of combined therapy.

Findings: EZH2 knockout increased FADS2 variant 1 (v1) and decreased variant 2/3 (v2/3) at both the mRNA and protein levels. EZH2 occupied the promoters of FADS2 v1 and v2/3, but the enrichment of H3K27me3 at the v2/v3 promoter was significantly lower than that at the v1 promoter. Furthermore, EZH2 inhibitors increased full-length FADS2 v1 expression and polyunsaturated fatty acid abundance in OC cells. A high FADS2 v1/v2 ratio was associated with poor prognosis in patients with OC. Targeting FADS2 with shRNAs or inhibitors reduced EZH2 levels and sensitised cancer cells to EZH2 inhibitors. The combined inhibition of EZH2 and FADS2 induced mitochondrial dysfunction and energy stress and synergised to reduce tumour growth in vitro and in vivo.

Interpretation: EZH2 inhibition increases the proportion of the full FADS2 isoform through differential regulation, which suggests a poor prognosis in OC patients. Simultaneous inhibition of EZH2 and FADS2 causes mitochondrial dysfunction and energy stress in OC, resulting in a more pronounced antitumour effect. Our study reveals a mechanistic connection between EZH2 and FADS2 and provides a potential therapeutic strategy for OC.

Funding: This work was supported by the Youth Program of the National Natural Science Foundation of China (No. 82203102, No. 82303801, No. 82203433), the National Key Research and Development Program of China (2022YFA1104001), the General Program of the National Natural Science Foundation of China (No. 82272745, No. 82072870, No. 81972966, No. 82373173, No. 82073057), and the Peking University Third Hospital Clinical Key Project (BYSY2022069, BYSY2022070, BYSYZD2023010).