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Small Molecule Protein Ligands as Modulators of Hypoxia-Inducible Transcription

小分子蛋白配体作为缺氧诱导转录的调节剂

基本信息

批准号：
8865244
负责人：
Bogdan Olenyuk
金额：
$ 33.3万
依托单位：
PROTEOGENOMICS RESEARCH INSTIT/SYS/ MED
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-05 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8865244
关键词：
Accounting Affinity Apoptotic Arts Binding Biological Assay Biological Factors Blood Vessels C-terminal CREB-binding protein CXCR4 gene Cell Culture Techniques Cell Nucleus Cells Characteristics Chemicals Chronic Complex DNA Sequence Development Disease Down-Regulation E1A-associated p300 protein EP300 gene Energy Metabolism Enzyme-Linked Immunosorbent Assay Event Fluorescence Polarization Future Gene Expression Gene Expression Profiling Genes Genetic Genetic Transcription Goals Growth HIF1A gene Human Hydroxylation Hypoxia Hypoxia Inducible Factor In Vitro Interleukin-2 LOX gene Laboratories Lead Length Libraries Ligand Binding Ligands Link Luciferases Malignant Neoplasms Mediating Membrane Proteins Methodology Methods Molecular Molecular Biology Mus Neoplasm Metastasis Organ Oxidation-Reduction Oxygen Pathway Analysis Pathway interactions Patients Pharmacology Play Polyubiquitination Process Prognostic Factor Property Proteins Proteomics Recruitment Activity Regulation Research Response Elements Selenium Signal Pathway Signal Transduction Site-Directed Mutagenesis Solid Solid Neoplasm Structure Surface Surface Plasmon Resonance Testing Therapeutic Titrations Transactivation Transcription Coactivator Transcriptional Activation Vascular Endothelial Growth Factors Work Xenograft Model analog angiogenesis base cancer therapy chaetocin chemical synthesis chetomin design efficacy testing genome-wide human CREBBP protein human genome sequencing hypoxia inducible factor 1 in vitro testing in vivo inhibitor/antagonist microcalorimetry mimetics neovascularization novel nuclear receptor coactivator 1 overexpression prevent protein protein interaction public health relevance receptor research study response small molecule therapy development tool transcription factor tumor tumor growth tumor progression tumor xenograft

项目摘要

DESCRIPTION (provided by applicant): Hypoxia, the state of reduced oxygen levels, triggers a multifaceted adaptive response mediated by oxygen- dependent transcription factors, termed hypoxia-inducible factors (HIFs). Among these, hypoxia-Inducible Factor 1 (HIF1) is the main regulator of oxygen-dependent transcription in a majority of organs and accounts for the increase in expression of hypoxia-inducible genes. HIF controls an expression of over 100 genes involved in angiogenesis, altered energy metabolism, anti-apoptotic, and pro-proliferative mechanisms. Elevated levels of HIF1 are an independent, prognostic factor for a diverse range of malignant neoplasms. We are developing small molecules chemical tools that provide exquisite control over transcriptional activation of hypoxia-inducible genes implicated in cancer progression. Specifically, we will target three key signaling events mediated by three distinct protein-protein interactions with unique synthetic ligands. Small molecules often interact non-specifically with large protein surfaces; and hence inhibition of protein-protein interactions with small molecules remains difficult. Nevertheless, we recently demonstrated that hypoxia-inducible signaling can be effectively modulated in vitro and in vivo by rationally designed dimeric epidithiodiketopiperazines (ETPs) - ligands that bind p300/CBP CH1 region, induce structural change to these domains and prevent it from interacting with HIF1 C-terminal transactivation domain. The specific hypothesis behind the proposed research is that tumor progression could be effectively down-regulated by targeting of HIF1 C-TAD in complex with the CH1 domain of the coactivator protein p300/CBP with these designed molecules. Specific Aim 1 focuses on synthesis of conformational mimetics of the dimeric epidithiodiketopiperazine (ETP) natural products, characterization of the ETPs binding properties via isothermal titration microcalorimetry (ITC), surface plasmon resonance (SPR), and fluorescence polarization competition experiments. In Specific Aim 2 we will evaluate the stability of dimeric ETPs in cell culture and test efficacy in inhibiting HIF-inducible transcription with luciferase assays, qRT-PCR, ELISA. In this Aim we will also investigate structural basis of the inhibition of HIF1-coactivator interactions by employing site- directed mutagenesis and structural NMR. In Specific Aim 3 we will explore mechanistic framework of ETPs with pathway-based RT2 Profiler assays, gene expression profiling, and pathway analysis. We will also conduct in vivo experiments in order to determine efficacy of the ETPs in our mouse tumor xenograft models. Combined the three aims will validate our hypothesis and could lead to novel, unique tools for dissecting the hypoxia-inducible signaling pathway in cancer and facilitate development of future therapeutics.