Adenovirus capable of expressing anti-cancer gene efficiently, regulated by miRNA and capable of specifically proliferating in glioma cells and application thereof

Abstract

The invention provides a recombinant adenovirus vector regulated by miRNA (micro Ribose Nucleic Acid) and capable of specifically proliferating in glioma cells. The recombinant adenovirus vector provided by the invention is characterized in that the target spot nucleotide sequence of at least one miRNA is inserted into the 3-untranslated region of at least one proliferation essential gene of the virus, the expression of miRNA is absent or reduced in glioma cells, but miRNA is normally expressed in nerve cells and glial cells so that the modified recombinant adenovirus specifically proliferate in the adenovirus in which the expression of miRNA is absent or reduced, but do not proliferate in normal cells in which the miRNA is normally expressed; and therefore, the recombinant adenovirus does not generate cytotoxicity to the normal cells.

Description

Field of invention: [0001] The present invention generally relates to recombinant adenoviral vectors. More specifically, the present invention relates to a recombinant adenoviral vector capable of specifically proliferating in glioma cells under the regulation of miRNA, as well as its construction method and application. Background technique: [0002] Gene therapy is a new type of treatment that can treat a variety of diseases, including cancer, genetic diseases, infectious diseases, cardiovascular diseases and autoimmune diseases. The past few years have seen tremendous advances in gene therapy around the world. As of September 2008, the total number of clinical trials has reached 1,432, and the first gene therapy drug-"Jinyousheng" has been independently developed by my country's Shenzhen Saibainuo Company and approved for marketing (see http: / / www.wiley. co.uk / genmed / clincal / ). [0003] Adenoviruses are currently one of the most common vectors for gene therapy, and ne...

AI Technical Summary

Problems solved by technology

[0005]However, there are still some problems with the transformed adenovirus vector. Taking tumor gene therapy as an example, firstly, the mechanism of tumor generation is very complex and highly heterogeneous , there are significant differences between patients, between tumors, and between different tumor cells of the same tumor

For example, although telomerase has been reported to be abnormally expressed in tumors, its expression varies greatly, and its expression in some patients, tumor types, or certain tumor cell populations is no different from that of normal cells, which makes telomeres In this case, the recombinant adenoviral vector that regulates E1A by the enzyme promoter cannot selectively proliferate efficiently in tumor cells, thus affecting the therapeutic effect

Second, physical factors in tumor tissue such as fibrosis, incorporation of normal cells, and necrotic areas may also limit the spread of adenoviruses

Third, the low expression of Coxsackie adenovirus receptors on the cell membrane of some tumors limits the infection efficiency of adenoviruses

Fourth, the patient's immune response limits the proliferation and spread of the adenovirus

At present, ONYX Pharmaceutical Company of the United States is trying to use adenovirus with E1b 55kD protein deletion alone (ONYX-015 to treat tumors, and its clinical effective rate is only 15-20% (Nemunaitis et al., 2000; US 5677178; US 5801029)

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