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 using in vitro and in vivo models including genetic mouse models, patient-derived xenograft models, and organoid models. The overall goals are to identify the novel hypoxia-dependent therapeutic vulnerabilities and ultimately to translate knowledge to cancer therapy. Currently, our research is supported by NCI, CPRIT, ACS, Welch, and Mary Kay foundation.
HIF and epigenetic reprogramming in human cancers.
We are currently studying the crosstalk between hypoxia/HIF and epigenetic regulators and their functional significance in tumor initiation and progression, immune evasion, as well as therapy resistance. We are also interested in the development of new targeted epigenetic therapies in human cancers. See some examples below.
1. How is RNA polymerase II regulated to control HIF transactivation in breast cancer (Chen Y et al., JCI 2018; Wang Y et al., Cancer Res 2020).
Hypoxia is a hallmark of tumor microenvironment and promotes tumor progression. Hypoxia-inducible factor (HIF) is a master transcription factor that controls hypoxia response in tumors. How the epigenetic regulator regulates HIF transcriptional activity to mediate tumor progression remains poorly understood. We recently identified several novel HIF-interacting proteins including ZMYND8, CHD4, and BRD4 in breast cancer cells (Chen Y et al. JCI 2018; Wang Y et al., Cancer Res 2020). In breast cancer cells under normoxia, CHD4 enrichment at HIF target gene promoters increases RNA polymerase II loading through p300. Hypoxia further promotes CHD4 binding to the chromatin via HIF-1/2alpha, where CHD4 in turn enhances recruitment of HIF-1alpha, leading to HIF target gene transcription. In contrast, ZMYND8 enhances elongation of HIF target genes by increasing recruitment of BRD4 and subsequent release of paused RNA polymerase II in breast cancer cells (Figure 1). ZMYND8 acetylation at lysine 1007 and 1034 by p300 is required for its binding to BRD4 and subsequent HIF activation. Loss of HIF-1/2alpha abolishes CHD4 or ZMYND8-mediated breast tumor growth in vitro and in mice. These findings uncover the epigenetic mechanisms underlying regulation of RNA polymerase II to promote HIF activation and HIF-mediated breast cancer progression, and discover possible molecular targets for the diagnosis and treatment of breast cancer.
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).
HIF and non-coding RNAs in human cancers
We are studying hypoxia-induced non-coding RNAs in human breast tumors and their roles in breast cancer progression. Recently we identified hypoxia-induced long non-coding RNA (lncRNA) transcriptome in triple-negative breast cancer cells (TNBCs) and characterized two novel lncRNAs RAB11B-AS1 and lncIHAT in breast cancer cells (Niu Y et al., Cancer Res 2020; Chen L et al., Mol Cancer Res 2021). RAB11B-AS1 is a natural lncRNA upregulated in human breast cancer and its expression is induced by hypoxia-inducible factor 2 (HIF-2), but not HIF-1, in response to hypoxia. RAB11B-AS1 enhanced the expression of angiogenic factors including VEGFA and ANGPTL4 in hypoxic breast cancer cells by increasing recruitment of RNA polymerase II. In line with increased angiogenic factors, conditioned media from RAB11B-AS1-overexpressing breast cancer cells promoted tube formation of human umbilical vein endothelial cells in vitro. Gain- and loss-of-function studies revealed that RAB11B-AS1 increased breast cancer cell migration and invasion in vitro and promoted tumor angiogenesis and breast cancer distant metastasis without affecting primary tumor growth in mice. Taken together, these findings uncover a fundamental mechanism of hypoxia-induced tumor angiogenesis and breast cancer metastasis (Figure 3).
Distinct to RAB11B-AS1, lncIHAT is abundantly expressed in human TNBC and predominantly induced by HIF-1 but not HIF-2. LncIHAT promoted TNBC cell survival in vitro and tumor growth and metastasis in mice through inducing its proximal neighboring genes PDK1 and ITGA6. Collectively, these findings uncovered lncIHAT as a new hypoxia-induced oncogenic cis-acting lncRNA in TNBC.
HIF and cancer metabolism
Hypoxia-inducible factors (HIFs) mediate metabolic reprogramming in response to hypoxia. We previously dissected a reciprocal regulation between HIF-1 and PKM2 in cancer cells. PKM2 is hydroxylated by PHD3 and promotes HIF-1 transactivation and reprograms glucose metabolism in cancer cells (Figure 4).
Recently, we found that hypoxia upregulates mRNA and protein levels of the branched-chain amino acid (BCAA) transporter LAT1 and the BCAA metabolic enzyme BCAT1, but not their paralogs LAT2-4 and BCAT2, in human glioblastoma (GBM) cell lines as well as primary GBM cells. Hypoxia-induced LAT1 protein upregulation is mediated by both HIF-1 and HIF-2 in GBM cells. Although both HIF-1alpha and HIF-2alpha directly bind to the hypoxia response element at the first intron of the human BCAT1 gene, HIF-1alpha is exclusively responsible for hypoxia-induced BCAT1 expression in GBM cells. Knockout of HIF-1alpha and HIF-2alpha significantly reduces glutamate labeling from BCAAs in GBM cells under hypoxia, which provides functional evidence for HIFs-mediated reprogramming of BCAA metabolism. Genetic or pharmacological inhibition of BCAT1 inhibits GBM cell growth under hypoxia. Together, these findings uncover a previously unrecognized HIFs-dependent metabolic pathway that increases GBM cell growth under conditions of hypoxic stress. (Zhang B, Chen Y, et al. Cell Mol Life Sci 2021; Peng H et al. Oncogene 2020).
Overall, we are interested in HIF-dependent metabolic reprogramming and metabolic therapy in human cancers.