成果報告書詳細
管理番号20130000000347
タイトル*平成24年度中間年報 「後天的ゲノム修飾のメカニズムを活用した創薬基盤技術開発」
公開日2014/8/20
報告書年度2012 - 2012
委託先名国立大学法人東京大学 エピゲノム技術研究組合
プロジェクト番号P10005
部署名バイオテクノロジー・医療技術部
和文要約
英文要約Title: Technology development for drug discovery platform based on the epigenetic mechanism (FY2010-FY2015) FY2012 Annual Report
 
The aim of this project is to establish novel technologies for the acceleration of diagnostic and drug discovery based on epigenomic modifications in cancer. The research has been promoted to identify the target molecules involved in regulating histone modification, methylation or de-methylation, and to validate the utility of the developed technologies. The major research results in FY2012 are summarized as follows; (1) Technology development for comprehensive analysis of epigenetic modifications: We optimized the whole-genome and base-resolution analysis of cytosine methylation, WGBS and PBAT. We began to establish high-resolution analysis of cytosine hydroxymethylation. We have also established the ChIP-seq method that allows us to examine protein binding profile from limited number of cells, as few as 10,000 cells. The benchtop sequencer was effective to accelerate the evaluation of these improved protocols. We improved the sensitivity of detection for combinatorial histone modifications using a newly-installed LC-MS and started to analyze co-immunoprecipitated histone tails. We also improved the accuracy of targeted quantitative proteomics using SRM. (2) Development of data mining platform to discover the relationship between epigenetic modifications and the disease: We have prepared the archive of frozen specimens and purified DNA and RNAs of total 90 human cancer tissues. We also established the direct xenografts and cell lines. Oncogenic roles of the histone modifiers were validated by genetically engineered mouse models. We performed transcriptome analysis using RNA-sequencing and selected the candidates of functional ncRNA. We further analyzed the direct binding and functional interaction between the ncRNA and chromatin complexes. We selected the candidate markers that are commonly methylated among multiple types of cancers or are aberrantly methylated in a cancer-type specific manner by using epigenotyping microarrays and went forward to diagnostic application using circulating blood DNA in patients. We identified the novel candidates for therapeutic drug targets by proteome analysis of chromatin complexes. (3) Exploratory experimental study: Using the technologies developed in the section 1, we have analyzed dynamics in combinatorial patterns of histone H4 tail modification. To improve accuracy, we have established the high sensitive quantification method and technology to detect multiple modification patterns simultaneously. We have established new systems to screen through libraries of natural compounds or peptides. We started to crystalize methylase-inhibitor complexes and analyze the structures of the complexes by X-ray crystallography.
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