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Characterization of Small Molecule ETS Transcription Factor Binders Nicole M. Martinez

Characterization of Small Molecule ETS Transcription Factor Binders Nicole M. Martinez Marius S. Pop and Levi A. Garraway Cancer Biology Program. DOI:10.1038/nrd2275. Targeted Therapy in Cancer. “Druggable” targets Obvious active site Kinases Other enzymes “Undruggable” targets

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Characterization of Small Molecule ETS Transcription Factor Binders Nicole M. Martinez

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  1. Characterization of Small Molecule ETS Transcription Factor Binders Nicole M. Martinez Marius S. Pop and Levi A. Garraway Cancer Biology Program

  2. DOI:10.1038/nrd2275 Targeted Therapy in Cancer • “Druggable” targets • Obvious active site • Kinases • Other enzymes • “Undruggable” targets • No obvious pocket

  3. Many “Driver” Cancer Proteins are Currently “Undruggable” • Example: Oncogenic Transcription Factors • ETS Transcription Factors ERG Translocated in >50% of prostate cancers ETV1 Otis Brawley, National Cancer Institute Prostate Tumor

  4. ETS Transcription Factor Role in Prostate Cancer /ETV1 doi:10.1038/nm0106-14

  5. Can we develop a therapeutic?

  6. ERG truncated ERG full ETV1 MITF αHA Small-Molecule Microarrays (SMMs) Lysates expressing target protein DMSO stock solutions ha.11 aMouse-IgG-Cy5 ETV1 protein-small molecule interaction on a microarray fluorescent features reveal putative binding interactions

  7. From Raw Data to Hits f = microarray featuremedian pixel intensity b = local backgroundmedian pixel intensity xcpd = f - b Z* 1.25 3.75 6.25 Overlay of GAL File cpd- µmock Z* = ( ) 0.96 cpd mock 1+

  8. ETV1 Selection of Hits ZScoreA ZScoreC ZScoreB CompositeZ CompositeZ

  9. ChemBank: Tool for Filtering Compounds http://chembank.broad.harvard.edu

  10. List of ETS “Hit” Compounds NPC: Natural Products and commercials (Including FDA approved drugs) library PDI: Psychiatric Disease Initiative compounds *10,800 compounds per library

  11. ERG Assess Inhibitory Capabilities by Luciferase Assays ETS Luciferase Fold rep rep + ERG + compound rep + ERG

  12. Lead Compounds Cpd1 Cpd2

  13. ERG Assess Inhibitory Capabilities by Luciferase Assays ETS Luciferase Fold rep rep + ERG + compound rep + ERG

  14. A B C D Ctrl DMSO ERG cpds MITF ETV1 A B C D Ctrl tERG cpds ETV1 MITF Ctrl ERG cpds Dud Compounds

  15. Conclusions • SMM allow us to find binders • Luciferase assays allow us to determine inhibitory capabilities • Future work • Surface Plasmon Resonance • Screen w/ more compounds

  16. Mentors Marius Pop, PhD Levi Garraway, MD, PhD Collaborators Angela Koehler, PhD Jason Fuller Summer Research Program in Genomics Shawna Young Lucia Vielma Bruce Birren, PhD Acknowledgements

  17. ETS Fusion Products • Exon 1 of TMPRSS2 with the beginning of exon 4 of ETV1 • Exons 1 and 2 of TMPRSS2 with the beginning of exon 4 of ETV1 • Exon 1 of TMPRSS2 with the beginning of exon 4 of ERG

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