成果報告書詳細
管理番号20120000001217
タイトル*平成23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」(エピゲノム技術研究組合、国立大学法人東京大学)
公開日2014/8/28
報告書年度2011 - 2011
委託先名エピゲノム技術研究組合 国立大学法人東京大学
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)FY2011 Annual Report

The aim of this project is to establish various 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, either methylation or de-methylation, and also to validate the utility of the newly developed technologies. The major research results in FY2011 are summarized as follows; (1)Technology development for comprehensive analysis of epigenetic modifications: We have performed the optimization of methylated DNA immunoprecipitation experiment with anti-methylcytosine antibody and verified that sufficient amount of methylated DNA product for MeDIP-sequencing analysis is available in case of one microgram of genomic DNA for a starting material. In addition, we established the hydroxymethylated DNA immunoprecipitation experiment for genome wide detection of hydroxymethylcytosine. We have also established the mini-scale ChIP-seq method that allows us to explore protein binding profile from limited number of cells (10,000 cells). We have developed a methodology to resolve temporal changes of many modification patterns for H4 tail with less than 1 ug histone protein. (2) Development of data mining platform to discover the relationship between epigenetic modifications and the disease: We have established the bank of total 60 cancer tissues originated from human gastric, lung or pancreatic cancers, continuously. we have embarked systematic selection for early detection of several types of cancers using epigenotyping microarrays. We selected the candidate markers which are commonly methylated among multiple types of cancers or are aberrantly methylated in a cancer-type specific manner. To discover targeting candidates for developing new molecular-targeting drugs, we generated the shRNA constructs for various histone or DNA modifiers and generated the knockdown cancer cells for them. We have found that the suppression of a demethylase, reduces the tumor forming ability in colon cancer cells. (3)Exprolatory studies for validation of the drug discovery technology and platform: For validation of the methods developed above, we analyzed histone H4 modification during cell cycle. In addition to drug screening for histone methylase with MALDI-TOF MS, we started to establish a new high throughput screening with alpha-screening method. We have determined more effective inhibitors by in vitro assay with MALDI-TOF MS among about 200 compounds selected in the 2nd in silico screening.
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