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.

1 . A lentiviral vector for introducing a gene into an airway epithelial stem cell, wherein the vector is pseudotyped with an RNA or DNA virus spike protein. 
     
     
         2 . The lentiviral vector of  claim 1 , wherein the RNA virus is an RNA virus that infects an airway tissue. 
     
     
         3 . The lentiviral vector of  claim 1 , wherein the RNA virus is a minus-strand RNA virus. 
     
     
         4 . The lentiviral vector of  claim 3 , wherein the minus-strand RNA virus is a paramyxovirus. 
     
     
         5 . The lentiviral vector of  claim 4 , wherein the paramyxovirus is Sendai virus. 
     
     
         6 . The lentiviral vector of  claim 3 , wherein the minus-strand RNA virus is an orthomyxovirus. 
     
     
         7 . The lentiviral vector of  claim 6 , wherein the orthomyxovirus is an influenza virus. 
     
     
         8 . The lentiviral vector of  claim 3 , wherein the minus-strand RNA virus is a filovirus. 
     
     
         9 . The lentiviral vector of  claim 8 , wherein the filovirus is Ebola hemorrhagic fever virus. 
     
     
         10 . The lentiviral vector of  claim 1 , wherein the RNA virus is a plus-strand RNA virus. 
     
     
         11 . The lentiviral vector of  claim 10 , wherein the plus-strand RNA virus is a coronavirus. 
     
     
         12 . The lentiviral vector of  claim 11 , wherein the coronavirus is SARS coronavirus. 
     
     
         13 . The lentiviral vector of  claim 1 , wherein the DNA virus is a DNA virus that infects an airway tissue. 
     
     
         14 . The lentiviral vector of  claim 13 , wherein the DNA virus is a baculovirus. 
     
     
         15 . The lentiviral vector of any one of  claims 1  to  14 , wherein the lentiviral vector is a recombinant simian immunodeficiency virus vector. 
     
     
         16 . The lentiviral vector of  claim 15 , wherein the recombinant simian immunodeficiency virus vector is derived from an agm strain. 
     
     
         17 . The lentiviral vector of  claim 15  or  16 , wherein the recombinant simian immunodeficiency virus vector is a self-inactivating vector. 
     
     
         18 . The lentiviral vector of any one of  claims 1  to  14 , wherein the lentiviral vector is an equine infectious anemia virus vector, human immunodeficiency virus-1 vector, human immunodeficiency virus-2 vector, or feline immunodeficiency virus vector. 
     
     
         19 . The lentiviral vector of any one of  claims 1  to  18 , which carries a foreign gene in an expressible state. 
     
     
         20 . The lentiviral vector of  claim 19 , wherein the foreign gene is a gene encoding a protein selected from the green fluorescent protein, beta-galactosidase, and luciferase. 
     
     
         21 . The lentiviral vector of  claim 19 , wherein the foreign gene is a gene encoding an inherent or acquired dysfunctional protein. 
     
     
         22 . The lentiviral vector of  claim 19 , wherein the foreign gene is a gene encoding an inherent or acquired dysfunctional cystic fibrosis (CF)-causing factor. 
     
     
         23 . The lentiviral vector of  claim 19 , wherein the foreign gene is a gene encoding an inherent or acquired dysfunctional CFTR protein. 
     
     
         24 . The lentiviral vector of  claim 19 , wherein the foreign gene is a gene encoding a protein having a therapeutic effect on cystic fibrosis. 
     
     
         25 . The lentiviral vector of  claim 19 , wherein the foreign gene is a gene encoding a protein that has become dysfunctional due to a genetic disease. 
     
     
         26 . The lentiviral vector of  claim 25 , wherein the protein that has become dysfunctional due to a genetic disease is a gene encoding CFTR. 
     
     
         27 . A method for introducing a gene into an airway epithelial stem cell, which comprises the step of contacting an airway epithelial cell with the lentiviral vector of any one of  claims 1  to  26 . 
     
     
         28 . An airway epithelial stem cell into which the lentiviral vector of any one of  claims 1  to  26  has been introduced. 
     
     
         29 . An agent for transferring a gene into an airway epithelial stem cell, which comprises the lentiviral vector of any one of  claims 1  to  26  as an active ingredient. 
     
     
         30 . The agent of  claim 29 , wherein the lung is used as a production tissue to provide a protein necessary for disease treatment. 
     
     
         31 . A therapeutic agent for genetic respiratory disease, which comprises the lentiviral vector of any one of  claims 1  to  26  as an active ingredient. 
     
     
         32 . The therapeutic agent of  claim 31 , wherein the genetic respiratory disease is cystic fibrosis.