GENE TRANSFER INTO AIRWAY EPITHELIAL STEM CELL BY USING LENTIVIRAL VECTOR PSEUDOTYPED WITH RNA VIRUS OR DNA VIRUS SPIKE PROTEIN
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 method of treating an individual having a genetic respiratory disease, comprising directly contacting airway cells of an individual in need thereof with a recombinant simian immunodeficiency virus (SIV) vector comprising a foreign gene operably linked to a promoter, the vector pseudotyped with spike proteins of Sendai virus envelope glycoproteins F and HN, wherein the vector integrates the foreign gene into an airway epithelial stem cell genome,
wherein the recombinant SIV vector comprises a chimeric sequence;
wherein the airway cells are not pretreated or pre-conditioned prior to being directly contacted with the vector, and
wherein the integrated foreign gene encodes a therapeutic agent that treats the genetic respiratory disease and is expressed in the airway epithelial stem cell for at least 160 days.
2 . The method of claim 1 , wherein the recombinant SIV vector is from an agm strain.
3 . The method of claim 2 , wherein the recombinant SIV vector is a self-inactivating vector in which the promoter sequence for the 3′ LTR U3 region is eliminated and/or substituted with another promoter sequence.
4 . The method of claim 1 , wherein the genetic respiratory disease is cystic fibrosis.
5 . The method of claim 1 , wherein the therapeutic agent encoded by the foreign gene is:
(a) a protein that in the individual has become inherently or acquiredly dysfunctional, resulting in the genetic disease;
(b) a cystic fibrosis-causing factor that in cystic fibrosis becomes inherently or acquiredly dysfunctional;
(c) a CFTR protein;
(d) a protein having a therapeutic effect on cystic fibrosis;
(e) a CFTR protein that is needed for the treatment of cystic fibrosis; or
(f) a CFTR protein, wherein expression of the CFTR protein supplements a deficient CFTR gene.
6 . The method of 5 , wherein the protein that has become inherently or acquiredly dysfunctional is CFTR.
7 . The method of claim 1 , wherein the foreign gene is expressed in the airway epithelial stem cell for at least 220 days or at least 360 days.
8 . The method of claim 1 , wherein the recombinant SIV vector is administered nasally.