| In recent years, gene therapy has made great progress. Around the worldthousands cases of gene therapy research have entered clinical trials. The typeof gene disease involved in congenital genetic diseases such as hemophilia andmany human high-risk diseases. The gene therapy ranges from the field ofgenetic disease extend to the tumor, cancer, cardiovascular diseases,neurological diseases, infectious diseases (including AIDS) and other fields.whatever potential bentfits of the gene treatment for the human health, genedrug delivery carrier limits gene therapy.Gene drug carriers evolved from naked DNA vectors, non-viral vectors tothe viral vectors. The most notable gene drug carrier is rAAV.Adeno-associated virus (Adeno-associated virus， AAV) belong to theparvovirus family, is a single-stranded non-enveloped virus, AAV has widehost range, non-pathogenic, can long-term stable express exogenous gene, etc.AAV was developed for gene therapy vectors-recombinant adeno-associatedvirus (recombinant adeno-associated virus，rAAV) vector, and is consideredone of the most promising gene therapy vectors. A large number of theadeno-associated virus clinical trials are still underway or in preparation, aremainly used in the treatment of muscle and eye diseases.For a long time researchers agreed that recombinant adeno-associated virusvector has the advantages of non-pathogenic and low immunogenicity. Earlypre-clinical studies involving rodents and primates also have never been foundto cause serious adverse reactions. Recent Hemophilia gene therapy clinicaltrials show that the AAV2starts CD8T cell-mediated immune response, Thetransduction of the carrier causes liver cell cytotoxic effect. Conducted by theUniversity of Pennsylvania to treat hemophilia, the rAAV gene medicationsphase I/II clinical studies have serious cellular immune response, clinicalresearch is forced to terminate. In addition to the immune response, rAAV gene durgs treat AAT gene defects, Parkinson’s disease in clinical trialsproved to be ineffective. More clinical research problems of the safety andefficacy of the rAAV vector, require more research of the higher efficiency ofrAAV.How to improve the expression efficiency of rAAV vector is an urgent andarduous task, RNAi for therapeutic gene bring new hope to the efficiency ofrAAV vector expression. The carrier and the therapeutic gene are two keys ingene drugs.In the current gene drug design ideas, carrier simply used to carrytherapeutic genes, rarely consider the effect of therapeutic genes on thevehicle itself.Some therapeutic genes play an anti-tumor role at the same time can alsoimprove the efficiency of the gene expression of the carrier. In2009, theJohnson team found that siRNA-mediated silencing of nucleolarphosphoprotein and inhibition of nucleolar protein, can increase the rAAVvector gene expression efficiency to a maximum of15times and30timesrespectively. In2011, the Wallen team filtered50siRNA which cansignificantly promote rAAV gene expression (8times higher than that ofrAAV gene expression) from5520siRNA library.The above studies not only proved that the use of RNAi technology cansignificantly improve the efficiency of rAAV vector expression but alsoprovide a good reference for our study. The same RNAi microRNAs mightalso can improve the ability of the gene expression efficiency of rAAVvectors.This paper will explore the relationship between miRNA and rAAV, nearly2000miRNA library in human umbilical vein endothelial cells (humanumbilical vein endothelial cell，HUVEC) would be screened to promote rAAVexpression efficiency. Further validation of these miRNA will be proceed,analysis of its potential target genes and signaling pathways will be studied,the comprehensive mechanism of rAAV will be further explored, this way willopen the door for gene therapy breakthrough. |
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