经鼻给予重组腺病毒携带TALEN序列原位敲除BDNF-AS基因治疗缺血性脑卒中

项目名称： 经鼻给予重组腺病毒携带TALEN序列原位敲除BDNF-AS基因治疗缺血性脑卒中

项目编号： No.81471181

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 姜永军

作者单位： 中国人民解放军东部战区总医院

项目金额： 70万元

中文摘要： 脑源性神经营养因子(BDNF)是促使缺血性卒中后脑组织自身修复的关键分子，但BDNF-AS(一种长链非编码RNA)却抑制其表达。从RNA水平抑制BDNF-AS由于基因不断转录形成新的RNA而变得困难或代价高昂。传统基因敲除因难以解决在体原位特异性敲除基因而无法用于临床，最近发展TALEN技术解决了这一难题。但经典TALEN技术因所含的FokI核酸内切酶需形成二聚体才起作用而需要细胞同时转染两个TALEN质粒，导致基因敲除效率低等。单链TALEN技术克服这一缺点。但血脑屏障阻止大分子进入脑组织内发挥作用，我们前述研究证实经鼻给予大分子能快速有效地进入脑组织。因此，本项目旨在开发重组腺病毒载体携带针对BDNF-AS的单链TALEN序列并经鼻给予治疗缺血性脑卒中。

中文关键词： 经鼻给药；脑缺血；TALEN；脑源性神经营养因子；长链非编码RNA

英文摘要： Brain-derived neurotrophic factor (BDNF) promotes brain self-repair after ischemic stroke while BDNF-AS, a long non-coding RNA, inhibits its expression. Because many oligonucleotides such as siRNA could only inhibit the expression of BDNF-AS temporarily, it is necessary to knock out the BDNF-AS gene to sustain the low copy of BDNF-AS. However, it is difficult to knock the specific gene in situ in vivo. Recently, transcription activator-like effector nuclease (TALEN) has been developed to knock out any specific gene due to its specific DNA binding domain and non-specific FokⅠendonuclease which only cut the DNA through the dimer. Hence, people needs to transfect two TALEN plasmids into the cells simultaneously which leads to low efficiency of transfection and severe adverse effects. A single chain carrying two Fok I domain could solve this problem. Moreover, the exogenous molecule especially the large weigh molecule cannot cross the intact blood brain barrier. Intranasal delivery of therapeutics could get into the central nervous system bypassing the blood brain barrier. The aim of this project is to develop a single chain TALEN targeting BDNF-AS gene and intranasal delivery of recombinant adenovirus vector carrying the single chain TALEN to treat experimental stroke.

英文关键词： intranasal delivery;ischemia;TALEN;BDNF;lncRNA