Development of new types of retrovirus vectors that modulate epigenetical regulation.

开发调节表观遗传调控的新型逆转录病毒载体。

基本信息

批准号：
17016015
负责人：
IBA Hideo
金额：
$ 29.57万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2009
项目状态：
已结题

项目摘要

In this project, we have designed, prepared and improved several retrovirus/lentivirus vectors that carry efficient transcriptional units for the expression of short hairpin (sh) RNA, miRNA, and the newly developed RNA decoy molecule that inhibits specific miRNA (TuD RNA). Using these vector systems, we have studied cancer epigenetics by concentrating on SWI/SNF chromatin remodeling complex.We have noticed that in human tumor cell lines that are deficient in Brm expression, expression of MLV and HIV vectors that were exogenously transduced into them are rapidly silenced stochastically. We finally demonstrated that Brm-type SWI/SNF complex is essential for the stable expression of retro/lentivirus.Brm is further shown to have antioncogenic potential because exogenous introduction of Brm reduces oncogenic potential of cell lines deficient in Brm expression. In all these cell lines examined, the functional Brm gene is present and is actively transcribed; Brm expression was suppressed at t … More he post-transcriptional level. We hypothesized that Brm is targeted by certain miRNAs and screened several miRNA candidates and finally have shown that miR-199a target Brm mRNA. Interestingly, all the cancer cell lines deficient in Brm have high levels of miR-199a, whereas in Brm expressing cells, miR-199a was marginally expressed.We further show these distinct expression patterns are resulted from double-negativefeedback regulation formed between Brm and miR-199a-5p/-3p via a transcription factor Egr1. We additionally have shown that miR-21 and its target NFIB, a negative transcriptional regulator, also forms a robust feedback regulation in cancer cell lines.We have also shown human requiem protein functions as an adaptor protein that links SWI/SNF complex and RelB/p52. We further showed that introduction of shRNA against REQ strongly suppresses anchourage-independent growth, but does not affect growth in monolayer culture at all in tumor cell lines such as Panc-1, in which non-canonical NFκB pathway is constitutively activated. Less

在这个项目中，我们设计，准备和改进了几个逆转录病毒/慢病毒载体，这些逆转录病毒/慢病毒载体具有有效的转录单元，用于表达短发夹（SH）RNA，miRNA和新开发的RNA诱饵分子，抑制特定的miRNA（TUD RNA）。使用这些载体系统，我们通过集中精力于SWI/SNF染色质重塑复合物来实现研究癌症表观遗传学。我们注意到，在BRM表达不足的人类肿瘤细胞系中，MLV和HIV载体的表达非常快，被极端翻译成它们的媒介物是快速的。我们最终证明，BRM型SWI/SNF复合物对于恢复/慢病毒的稳定表达至关重要。BRM进一步证明具有抗源性潜力，因为外源性引入BRM降低了BRM表达缺乏细胞系的致癌潜力。在所有检查的细胞系中，功能性BRM基因存在并积极转录。 BRM表达在T ...更多他的转录后级别被抑制。我们假设BRM是由某些miRNA靶向的，并筛选了几个miRNA候选物，并最终表明miR-199a靶标BRM mRNA。有趣的是，所有缺乏BRM的癌细胞系具有高水平的miR-199a，而在BRM表达细胞中，miR-199a的缘故均略有表达。我们进一步显示，这些独特的表达模式是由BRM和miR-199a-5p/-3p通过转录因子Egr1所形成的双模含量反馈调节导致的。我们还表明，miR-21及其靶标NFIB（一种负转录调节剂）也形成了癌细胞系中强大的反馈调节。我们还显示了人类所需的蛋白蛋白作为一种将SWI/SNF复合物和RELB/p52连接起来的衔接蛋白。我们进一步表明，针对REQ的SHRNA的引入强烈抑制与锚定的非依赖性生长，但在诸如Panc-1之类的肿瘤细胞系中根本不影响单层培养物的生长，在该肿瘤细胞系中，非典型的NFκB途径是由构造激活的。较少的

项目成果

期刊论文数量（90）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cdx2 and the Brm-type SWI/SNF complex cooperatively regulate villin expression in gastrointestinal cells

Cdx2和Brm型SWI/SNF复合物协同调节胃肠细胞绒毛蛋白表达

DOI：
发表时间：
2009
期刊：
Experimental Cell Research 315
影响因子：
0
作者：
Yamamichi;N.;Inada;K.;Furukawa;C.;Sakurai;K.;Tando;T.;Ishizaka;A.;Haraguchi;T.;Mizutani;T.;Fujishiro;M.;Shimomura;R.;Oka;M.;Ichinose;M.;Tsutsumi;Y.;Omata;M.;Iab;H.
通讯作者：
H.

p54nrbとSWI/SNF複合体の相互作用とその生化学的意義

p54nrb与SWI/SNF复合物的相互作用及其生化意义