Gene transfer into airway epithelial stem cell by using lentiviral vector pseudotyped with RNA virus or DNA virus spike protein

Abstract

The present inventors successfully introduced genes into stem cells of airway epithelial tissues using simian immunodeficiency virus vectors pseudotyped with F and HN, which are envelope glycoproteins of Sendai virus. Gene transfer into airway epithelial tissue stem cells using a vector of the present invention is useful for gene therapy of genetic respiratory diseases such as cystic fibrosis. Furthermore, it is possible to select respiratory organs such as the lungs as production tissues for providing proteins that are deficient due to genetic diseases.

Description

TECHNICAL FIELD[0001]The present invention relates to lentiviral vectors pseudotyped with RNA or DNA virus spike proteins for introducing genes into airway epithelial stem cells.BACKGROUND ART[0002]Cystic fibrosis (CF) is the most common lethal autosomal recessive disorder among Caucasians. It is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Such mutations prevent ion transfer across the airway epithelium, which leads to mucosal thickening and attachment of bacteria, ultimately causing the disorder. Therefore, airway epithelial cells are important targets for gene therapy.[0003]For long-term expression of transgenes, it is important to introduce genes into airway epithelial progenitor cells including stem cells. “Stem cells” refers to undifferentiated cells which are multipotent and capable of self-renewal. In the case of the airway, stem cells are present near the ductal epithelia of the sub-mucosal glands and the basal cells of the basement membran...

AI Technical Summary

Benefits of technology

[0013]As described above, the present inventors developed vectors produced by pseudotyping simian immunodeficiency virus vectors with glycoproteins F and HN which are spike proteins of the airway-infecting Sendai virus; and developed methods of using these vectors for efficiently introducing genes into airway epithelial stem cells without preconditioning, from the apical side through the mucus layer. By using glycoproteins of airway-infecting viruses as envelope proteins, it became possible to introduce genes into airway epithelial cells as well as stem cells including progenitor cells. The technique for introducing genes into stem cells using the vectors of the present invention enables stable maintenance of gene expression, well beyond the survival time of epithelial cells.

Problems solved by technology

Such mutations prevent ion transfer across the airway epithelium, which leads to mucosal thickening and attachment of bacteria, ultimately causing the disorder.

In general, since epithelial stem cells are very few in number and exist in isolated locations in the sub-mucosal glands, it is difficult to adopt the gene transfer approach and such.

Furthermore, it is known that airway epithelial cells including stem cells are tissues where gene transfer is very difficult due to the presence of the mucin layer and mucus.

Since there are no virus receptors on the apical surface of airway epithelial cells, there are hardly any vectors that can be efficiently transduced into intact airway epithelium.

However, clinically it may not be possible to use treatment with these chemical substances.

However, such treatments (preconditioning) are not desirable for clinical application to humans.

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