There is an immediate need for pharmacological agents to potently and effectively modulate specific epigenetic enzymes and effector molecules. We have designed, established and successfully used a mammalian cell- based assay to identify chemical modulators of epigenetic pathways using a broad rather than a target specific approach to maximize the mechanistic diversity of assay actives. Using this assay we have recently conducted quantitative HTS using a titration-response method to evaluate more than 300,000 diverse compounds at multiple doses. We have identified and independently confirmed more than 70 actives. Evaluation of the anti- cancer potential of one of these hits, uncovered that this drug selectively targets lung cancer cells blocking their growth and triggering their cell death, without affecting normal human bronchial epithelial cells. This compound alters transcriptional pathways and affects epigenetic modifications of histones. In vivo, it shows anti-tumor activity in mouse xenografts. Our goal now is to characterize this top hit, which we call Drug 4, determining the epigenetic events and genomic sites it modifies, defining the mechanism of its anti-growth/anti- cancer activity and identifying the cellular proteins it targets, and in this way, to set up a pipeline through which we can characterize our other hits in future studies. We will use pairs of matched cell lines derived from the normal and the cancerous lung of patients to characterize the effects of this compound on the physiological vs. the pathological epigenomic/transcriptional landscape through the following aims: 1.Measure the effects of Drug 4 in normal vs. cancer cells on the expression level of genes that control cell proliferation pathways and evaluate epigenetic marks in cancer cells and in tumors at a subset of these loci selectively altered by Drug 4 in cancer. We will measure levels of genes that regulate cell growth/cell death pathways to identify those selectively affected by our drug in cancer but not in normal cells. We will then define drug 4-induced changes in epigenetic marks at the promoters of a subset of these genes. 2.Define the mechanism of Drug 4 induced growth inhibition/cancer cell death, in cells and in tumors. We will investigate if the cancer-specific cell cycle arrest &cell death triggered by Drug 4 depend on the cyclin D3/Rb pathway &the Egln3/KIF1B pathways, respectively, as suggested by preliminary results. 3.Identify the cellular targets of our compound through candidate molecule and unbiased approaches. We will determine the effect of Drug 4 on the activity of purified epigenetic enzymes and effector molecules to identify compound targets. We will perform biotinylated-drug pull downs and siRNA screens if needed.
These aims will help us elucidate the mechanism of action of Drug 4, and in particular, its selectivity against cancer. The studies proposed here will also lay the foundation for the future characterization and development of the other potential epigenetic modulators we have already identified. These compounds will have significant impact on basic research and on clinical therapeutic development for many pathologies, including cancer.

Public Health Relevance

We have identified a drug that shuts down genes that foster uncontrolled cell growth and turns on genes that slow down tumor growth. This compound shows the ability to specifically kill cancer cells and to decrease tumor size in mice, without affecting normal cells. Here, we propose to define how this drug works and to further develop it for therapeutic applications.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA125269-03
Application #
8215846
Study Section
Basic Mechanisms of Cancer Therapeutics Study Section (BMCT)
Program Officer
Arya, Suresh
Project Start
2010-03-01
Project End
2015-01-31
Budget Start
2012-02-01
Budget End
2013-01-31
Support Year
3
Fiscal Year
2012
Total Cost
$319,356
Indirect Cost
$118,081
Name
University of Texas Sw Medical Center Dallas
Department
Pharmacology
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
Bayo, Juan; Tran, Tram Anh; Wang, Lei et al. (2018) Jumonji Inhibitors Overcome Radioresistance in Cancer through Changes in H3K4 Methylation at Double-Strand Breaks. Cell Rep 25:1040-1050.e5
Dalvi, Maithili P; Wang, Lei; Zhong, Rui et al. (2017) Taxane-Platin-Resistant Lung Cancers Co-develop Hypersensitivity to JumonjiC Demethylase Inhibitors. Cell Rep 19:1669-1684
Martinez, Elisabeth D; Gazdar, Adi F (2016) Inhibiting the Jumonji family: a potential new clinical approach to targeting aberrant epigenetic mechanisms. Epigenomics 8:313-6
Trošt, Nuša; Peña-Llopis, Samuel; Koirala, Sajjan et al. (2016) ?Klotho is a novel marker and cell survival factor in a subset of triple negative breast cancers. Oncotarget 7:2611-28
Karamysheva, Zemfira; Díaz-Martínez, Laura A; Warrington, Ross et al. (2015) Graded requirement for the spliceosome in cell cycle progression. Cell Cycle 14:1873-83
Bayo, Juan; Dalvi, Maithili P; Martinez, Elisabeth D (2015) Successful strategies in the discovery of small-molecule epigenetic modulators with anticancer potential. Future Med Chem 7:2243-61
Schneider, Philipp; Bayo-Fina, Juan Miguel; Singh, Rajeev et al. (2015) Identification of a novel actin-dependent signal transducing module allows for the targeted degradation of GLI1. Nat Commun 6:8023
Chen, Yulong; Terajima, Masahiko; Yang, Yanan et al. (2015) Lysyl hydroxylase 2 induces a collagen cross-link switch in tumor stroma. J Clin Invest 125:1147-62
Díaz-Martínez, Laura A; Karamysheva, Zemfira N; Warrington, Ross et al. (2014) Genome-wide siRNA screen reveals coupling between mitotic apoptosis and adaptation. EMBO J 33:1960-76
Huffman, Kenneth; Martinez, Elisabeth D (2013) Pre-clinical studies of epigenetic therapies targeting histone modifiers in lung cancer. Front Oncol 3:235

Showing the most recent 10 out of 12 publications