Model systems for the investigation of DNA methylation and drug repurposing

用于研究 DNA 甲基化和药物再利用的模型系统

基本信息

批准号：
8679870
负责人：
John Russell Bracht
金额：
$ 14.96万
依托单位：
AMERICAN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8679870
关键词：
Aberrant DNA Methylation Accounting Address Applications Grants Applied Research Azacitidine Basic Science Biological Assay Biological Models Biology Cancer Biology Cancerous Cells Chemicals Chromosome abnormality Chromosomes Clinical Clinical Research Clinical Trials Clonal Expansion Complex Cytosine DNA DNA Methylation DNA Sequence DNA Sequence Rearrangement DNA-PKcs Data Decitabine Deposition Development Disease Elements Enhancers Epigenetic Process Eukaryota Event FDA approved Faculty Genetic Genome Genomic Instability Goals Grant Health Homologous Gene Human Human Biology In Vitro Investigation K-Series Research Career Programs Laboratories Left Libraries Link Manuscripts Measures Methylation Methyltransferase Mouse Cell Line Mus Noise Nuclear Organism Oxytricha Paper Pathway interactions Pharmaceutical Preparations Phenotype Play Positioning Attribute Preclinical Drug Evaluation Premalignant Cell Process Property Proteins Publications Publishing RNA Splicing Repetitive Sequence Research Role Running Scientific Advances and Accomplishments Staging System Techniques Technology Testing Therapeutic Time Tumor Suppressor Genes Tumor Suppressor Proteins Work Writing Yeast Model System Yeasts cancer cell career cell transformation clinical application drug discovery fly follow-up high throughput screening inhibitor/antagonist innovation interest meetings member metaplastic cell transformation microbial new technology next generation novel screening small molecule small molecule libraries tumor tumorigenesis validation studies

项目摘要

DESCRIPTION (provided by applicant): In this proposal, we will use two biological models to investigate DNA methylation, which plays key roles in oncogenesis yet is absent in many model systems. One system we utilize is the ciliate Oxytricha trifallax, which was only recently discovered to use cytosine methylation in a remarkable genome rearrangement process. Aim 1 is a mechanistic investigation of a novel methyltransferase complex biochemically purified from Oxytricha nuclear lysate. The role of one complex member, a DNA-PKcs homolog, in detecting aberrant chromosomes and guiding DNA methylation is a particular focus. Aim 2 is a combined mouse cell line (10T1/2) and Oxytricha screen of FDA-approved clinical compounds to identify those with methylation-modulating properties. Both Oxytricha and mouse 10T1/2 cells provide robust DNA methylation-driven phenotypes, allowing rapid screening for novel active compounds. Given the lack of DNA methylation in worms, flies, and yeast, these model systems provide ideal platforms for rapid scientific advance and for drug screening and repurposing efforts. Immediate and Long-term Career Goals My ultimate career goal is to be an innovative tenure-track faculty member who makes important contributions to basic and clinical research in the exciting field of epigenetics. Central to this goal is a shift from purely basic research to a combination of basic and applied research centered on the unique biology of Oxytricha and other model systems. The proposed research elaborates the advantages of Oxytricha into both clinical and basic research trajectories, and also rekindles an older mouse cell line, 10T1/2, for methylation drug screening, a novel application. This work will meet a deep need for model systems to study DNA methylation with clinical applications. In order to transition into an independent faculty position, I will perform the basic research described in Specific Aim 1 immediately, culminating in the publication of a manuscript describing the novel methylation machinery in Oxytricha. Year 2 will be dedicated to performing the chemical screen for methylation inhibitors (Aim 2) and writing an R01 grant application with the resultant data. I hope to have the screen finished by the end of Year 2, leaving Year 3 for a methylation enhancer screen and follow-up on mechanisms of action of the interesting hits, validation studies, and manuscript writing. This Career Development Award will allow the protected time necessary to establish publication and grant support necessary for a fast start in running my own laboratory.

描述（由申请人提供）：在本提案中，我们将使用两种生物模型来研究 DNA 甲基化，DNA 甲基化在肿瘤发生中发挥关键作用，但在许多模型系统中并不存在。我们使用的一个系统是纤毛虫 Oxytricha trifallax，它最近才被发现在显着的基因组重排过程中使用胞嘧啶甲基化。目标 1 是对从尖毛虫核裂解液中生物化学纯化的新型甲基转移酶复合物进行机理研究。一个复杂成员（DNA-PKcs 同源物）在检测异常染色体和指导 DNA 甲基化中的作用是一个特别关注的焦点。 Aim 2 是结合小鼠细胞系 (10T1/2) 和 FDA 批准的临床化合物的 Oxytricha 筛选，以鉴定具有甲基化调节特性的化合物。 Oxytricha 和小鼠 10T1/2 细胞均提供强大的 DNA 甲基化驱动表型，从而可以快速筛选新型活性化合物。鉴于蠕虫、果蝇和酵母中缺乏 DNA 甲基化，这些模型系统为快速科学进步以及药物筛选和重新利用工作提供了理想的平台。近期和长期职业目标我的最终职业目标是成为一名创新型终身教授，为激动人心的表观遗传学领域的基础和临床研究做出重要贡献。这一目标的核心是从纯粹的基础研究转向以尖毛虫和其他模型系统的独特生物学为中心的基础研究和应用研究相结合。拟议的研究将 Oxytricha 的优势阐述到临床和基础研究轨迹中，并重新点燃了较老的小鼠细胞系 10T1/2，用于甲基化药物筛选，这是一种新的应用。这项工作将满足对模型系统研究 DNA 甲基化及其临床应用的深刻需求。为了过渡到独立的教职职位，我将立即进行具体目标 1 中描述的基础研究，最终发表一篇描述 Oxytricha 中新型甲基化机制的手稿。第 2 年将致力于对甲基化抑制剂进行化学筛选（目标 2），并使用所得数据撰写 R01 拨款申请。我希望在第二年年底完成筛选，第三年进行甲基化增强剂筛选，并跟踪有趣的命中的作用机制、验证研究和手稿写作。该职业发展奖将为建立出版物提供必要的受保护时间，并为快速启动我自己的实验室提供必要的支持。