Award Title: Biochemical and Cellular Characterisation of Histone Demethylases
Funder: Cancer Research UK

Abstract: Post-oligomerisation modifications to chromatin are crucially important in transcriptional and epigenetic regulation. Abnormal modification patterns of DNA methylation histones are implicated in the onset and progression of multiple cancers. Small-molecules targeting chromatin modifying enzymes for cancer treatment are in clinical use. Academic and industrial researchers are progressing N-methyl lysine histone demethylases (KDMs) as cancer targets, with orally active compounds being reported; however, progress is limited by incomplete knowledge of KDM functions. Our ongoing CRUK work is exploring JmjC-KDM biochemistry and functions involving modifications to histones/other proteins so informing on roles of epigenetics in cancer and enabling drug discovery.     Aims  We are carrying out comprehensive functional assignments of human JmjC KDMs /hydroxylases, at biochemical and cellular levels so enabling their roles in cancer to be placed on a sound biochemical basis.    Methods  WF1. Biochemical characterization and functional assignments of JmjC-KDMs. To inform on roles of epigenetics in cancer and enable drug discovery, we will continue defining biochemical and cellular functions of JmjC-KDMs, building on recent breakthroughs. Areas of focus include (i) dual functionalities of arginine and lysine demethylation/completely novel JmjC catalysed modifications, (ii) KDM mediated acetylation and methylation cross-talk. WF2. Mechanistic studies and enabling development of new generation KDM inhibitors. Due to highly conserved active sites in JmjC-KDMs, achieving inhibitor selectivity is challenging, as evidenced by lack of high-quality chemical functional probes. Using combined chemical, bio-chemical/physical approaches, we will develop new strategies targeting JmjC KDMs involving allosteric/non-catalytic domains/substrate-binding sites, activity modulation (rather than ablation), and structure-based design of non-metal chelating scaffolds. WF3. Investigating the cellular roles of JmjC-KDMs and histone methylation in tumorigenesis/cancer. We will investigate the cellular effects of methylation cross-talk/novel modifications on histones, with respect to roles of KDMs, methyltransferases and acetyltransferases, and analyse links to oncogenic expression profiles. Building on work on transcription factors we will explore cellular roles of JmjC enzymes in ‘epigenetic memory’ and drug resistance.      How research results will be used  Our functional assignments will be used both by the large research community working on relationships between altered mechanisms of epigenetic regulation and cancer. They will be enabling to academic and pharmaceutical scientists pursuing JmjC-KDMs as cancer targets– an understanding of function is essential both for establishing optimal screens and interpreting the results of inhibitors in cells and animal models/clinical trials. We will work with our network of collaborators to disseminate results for use in studies aimed at validating JmjC enzymes as cancer targets.

Lay Abstract: Post-oligomerisation modifications to chromatin are crucially important in transcriptional and epigenetic regulation. Abnormal modification patterns of DNA methylation histones are implicated in the onset and progression of multiple cancers. Small-molecules targeting chromatin modifying enzymes for cancer treatment are in clinical use. Academic and industrial researchers are progressing N-methyl lysine histone demethylases (KDMs) as cancer targets, with orally active compounds being reported; however, progress is limited by incomplete knowledge of KDM functions. Our ongoing CRUK work is exploring JmjC-KDM biochemistry and functions involving modifications to histones/other proteins so informing on roles of epigenetics in cancer and enabling drug discovery.     Aims  We are carrying out comprehensive functional assignments of human JmjC KDMs /hydroxylases, at biochemical and cellular levels so enabling their roles in cancer to be placed on a sound biochemical basis.    Methods  WF1. Biochemical characterization and functional assignments of JmjC-KDMs. To inform on roles of epigenetics in cancer and enable drug discovery, we will continue defining biochemical and cellular functions of JmjC-KDMs, building on recent breakthroughs. Areas of focus include (i) dual functionalities of arginine and lysine demethylation/completely novel JmjC catalysed modifications, (ii) KDM mediated acetylation and methylation cross-talk. WF2. Mechanistic studies and enabling development of new generation KDM inhibitors. Due to highly conserved active sites in JmjC-KDMs, achieving inhibitor selectivity is challenging, as evidenced by lack of high-quality chemical functional probes. Using combined chemical, bio-chemical/physical approaches, we will develop new strategies targeting JmjC KDMs involving allosteric/non-catalytic domains/substrate-binding sites, activity modulation (rather than ablation), and structure-based design of non-metal chelating scaffolds. WF3. Investigating the cellular roles of JmjC-KDMs and histone methylation in tumorigenesis/cancer. We will investigate the cellular effects of methylation cross-talk/novel modifications on histones, with respect to roles of KDMs, methyltransferases and acetyltransferases, and analyse links to oncogenic expression profiles. Building on work on transcription factors we will explore cellular roles of JmjC enzymes in ‘epigenetic memory’ and drug resistance.      How research results will be used  Our functional assignments will be used both by the large research community working on relationships between altered mechanisms of epigenetic regulation and cancer. They will be enabling to academic and pharmaceutical scientists pursuing JmjC-KDMs as cancer targets– an understanding of function is essential both for establishing optimal screens and interpreting the results of inhibitors in cells and animal models/clinical trials. We will work with our network of collaborators to disseminate results for use in studies aimed at validating JmjC enzymes as cancer targets.

Research Activity Codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health Categories: CANCER AND NEOPLASMS