Retroviral vector targeting human cells via c-Kit-stem cell factor interaction.
Targeted gene transfer into hematopoietic stem cells by retroviral vectors would greatly facilitate the development of in vivo strategies for stem cell gene therapy. We engineered a recombinant retroviral vector that can target human cells expressing a c-Kit receptor via a ligand-receptor interaction. The ecotropic (Moloney murine leukemia virus) envelope protein was modified by insertion of a sequence encoding the N-terminal 161 amino acids of murine stem cell factor (mSCF), the ligand for murine c-Kit. The chimeric envelope protein was correctly processed and incorporated into viral particles as efficiently as the wild-type envelope protein. Virions pseudotyped with the chimeric envelope proteins bound to 293 cells expressing murine c-Kit (293KIT) preferentially; however, they could not transduce any c-Kit-positive cells under conventional conditions. They could transduce 293KIT cells in the presence of chloroquine, and HEL cells expressing human c-Kit on a fibronectin fragment (CH296)-coated dish. The fact that recombinant mSCF in the medium at the time of transduction greatly reduced the efficiency of both gene deliveries implies that the vector utilized the mSCF-c-Kit interaction for the initial step of transduction in either case. The vector may prove useful for targeting cells expressing c-Kit on their surface.
Yajima
T
T
The Heart Institute of Japan, Tokyo Women's Medical College.
Kanda
T
T
Yoshiike
K
K
Kitamura
Y
Y
D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D004274:DNA, Recombinant / Q000138:chemical synthesis; D016503:Drug Delivery Systems; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008979:Moloney murine leukemia virus; D016254:Mutagenesis, Insertional; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000378:metabolism; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics; D019089:Stem Cell Factor / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics
D004274:DNA, Recombinant; D008024:Ligands; D011993:Recombinant Fusion Proteins; D015682:Retroviridae Proteins, Oncogenic; D019089:Stem Cell Factor; D014759:Viral Envelope Proteins; D002738:Chloroquine; D019009:Proto-Oncogene Proteins c-kit