The central theme of our research is to understand the role of hypoxia-inducible factor (HIF) in tumor initiation and progression at the molecular and cellular levels. The overall goals are to identify the novel hypoxia-dependent therapeutic vulnerabilities and ultimately to translate knowledge to breast cancer therapy.

Epigenetic regulation of HIF in human cancers.

We are currently studying how HIF is activated by epigenetic regulators and their functional significance in cancer progression and metastasis.

1. The epigenetic reader ZMYND8 is a primary HIF coactivator in breast cancer (Chen Y et al., JCI 2018).

We found that ZMYND8 is induced by HIF-1 and HIF-2 in breast cancer cells and also upregulated in human breast tumors, and is correlated with poor survival of breast cancer patients. Genetic deletion of ZMYND8 decreases breast cancer cell colony formation, migration, and invasion in vitro, and inhibits breast tumor growth and metastasis to the lungs in mice. The ZMYND8’s oncogenic effect in breast cancer requires HIF-1 and HIF-2. We further showed that ZMYND8 interacts with HIF-1α and HIF-2α and enhances elongation of the global HIF-induced oncogenic genes by increasing recruitment of BRD4 and subsequent release of paused RNA polymerase II in breast cancer cells. ZMYND8 acetylation at lysine 1007 and 1034 by p300 is required for HIF activation and breast cancer progression and metastasis. These findings uncover a primary epigenetic mechanism of HIF activation and HIF-mediated breast cancer progression, and discover a possible molecular target for the diagnosis and treatment of breast cancer (Figure 1).


Figure 1. The molecular mechanism underlying ZMYND8-mediated HIF activation and breast cancer progression and metastasis

2. The methyltransferases G9a/GLP are the negative HIF-1 coregulators in GBM (Bao L et al., NAR 2018)

We found that the lysine methyltransferases G9a and GLP directly bound to the α subunit of HIF-1 (HIF-1α) and catalyzed mono- and di-methylation of HIF-1α at lysine (K) 674 in vitro and in vivo. K674 methylation suppressed HIF-1 transcriptional activity and expression of its downstream target genes PTGS1, NDNF, SLC6A3, and Linc01132 in human glioblastoma U251MG cells. Inhibition of HIF-1 by K674 methylation is due to reduced HIF-1α transactivation domain function but not increased HIF-1α protein degradation or impaired binding of HIF-1 to hypoxia response elements. K674 methylation significantly decreased HIF-1-dependent migration of U251MG cells under hypoxia. Importantly, we found that G9a was downregulated by hypoxia in glioblastoma, which was inversely correlated with PTGS1 expression and survival of patients with glioblastoma. Therefore, our findings uncover a hypoxia-induced negative feedback mechanism that maintains high activity of HIF-1 and cell mobility in human glioblastoma (Figure 2).

Figure 2. The molecular mechanism underlying G9a/GLP-mediated HIF-1 repression in GBM cells.

HIF and cancer metabolism

We are currently studying the HIF-dependent metabolic pathways and their roles in cancer progression. We previously showed that PKM2 gene transcription is activated by HIF-1. PKM2 interacts directly with the HIF-1α subunit and promotes transactivation of HIF-1 target genes by enhancing HIF-1 binding and p300 recruitment to hypoxia response elements, whereas PKM1 fails to regulate HIF-1 activity. Interaction of PKM2 with prolyl hydroxylase 3 (PHD3) enhances PKM2 binding to HIF-1α and PKM2 coactivator function. Mass spectrometry and anti-hydroxyproline antibody assays demonstrate PKM2 hydroxylation on proline-403/408. PHD3 knockdown inhibits PKM2 coactivator function, reduces glucose uptake and lactate production, and increases Oconsumption in cancer cells. Thus, PKM2 participates in a positive feedback loop that promotes HIF-1 transactivation and reprograms glucose metabolism in cancer cells (Figure 3). 

Figure 3. The molecular mechanism underlying PKM2-mediated HIF activation in cancer cells