Gene Therapy

Abstract

Use of cyclosporin A (CsA) or a derivative thereof for increasing the efficiency of transduction of an isolated population of human haematopoietic stem and/or progenitor cells by a vector derived from HIV-1, HIV-2, FIV, BIV, EIAV, CAEV or visna lentivirus.

1 - 25 . (canceled) 
     
     
         26 . A method of transducing a population of human haematopoietic stem and/or progenitor cells comprising the steps of:
 a) contacting the population of cells with cyclosporin A (CsA); and   b) transducing the population of cells with a HIV-1, HIV-2, FIV, BIV, EIAV, CAEV or visna lentiviral vector;   wherein transduction efficiency is increased in comparison to transduction in the absence of the CsA.   
     
     
         27 . The method of  claim 26 , wherein steps (a) and (b) are carried out ex vivo or in vitro. 
     
     
         28 . The method of  claim 26 , wherein the percentage of haematopoietic stem and/or progenitor cells transduced by the vector is increased and/or the vector copy number per cell is increased. 
     
     
         29 . The method of  claim 26 , wherein the vector is a HIV-1 vector. 
     
     
         30 . The method of  claim 26 , wherein the vector is a wild-type HIV-1 vector. 
     
     
         31 . The method of  claim 26 , wherein the vector is an integration-defective lentiviral vector (IDLV). 
     
     
         32 . The method of  claim 26 , wherein the vector comprises a capsid comprising glutamic acid at a position analogous to amino acid 92 and/or threonine at a position analogous to amino acid 88 of SEQ ID NO: 1, wherein the N-terminal proline of SEQ ID NO: 1 is assigned to be position 1. 
     
     
         33 . The method of  claim 26 , wherein the CsA is at a concentration of about 5-15 μM. 
     
     
         34 . The method of  claim 26 , wherein the population of cells is contacted with CsA in combination with rapamycin. 
     
     
         35 . The method of  claim 26 , wherein the population of cells is contacted with CsA in combination with 3-methyladenine (3-MA). 
     
     
         36 . The method of  claim 26 , wherein the population of haematopoietic stem and/or progenitor cells is obtained from mobilised peripheral blood, bone marrow or umbilical cord blood. 
     
     
         37 . The method of  claim 26 , which includes a further step of enriching the population for haematopoietic stem and/or progenitor cells. 
     
     
         38 . The method of  claim 37 , wherein the step of enriching the population for haematopoietic stem and/or progenitor cells is carried out before contacting the population of cells with CsA. 
     
     
         39 . The method of  claim 37 , wherein the step of enriching the population for haematopoietic stem and/or progenitor cells is carried out after transducing the population of cells with the vector. 
     
     
         40 . The method of  claim 26 , which further includes a wash step. 
     
     
         41 . The method of  claim 40 , wherein the wash step is carried out after transducing the population of cells with the vector. 
     
     
         42 . The method of  claim 40 , wherein the wash step is carried out after contacting the population of cells with CsA and before transducing the cells with the vector. 
     
     
         43 . A method of gene therapy comprising the steps of:
 a) transducing a population of human haematopoietic stem and/or progenitor cells according to the method of  claim 26 ; and   b) administering the transduced cells to a subject.   
     
     
         44 . The method of  claim 43 , wherein the transduced cells are administered to a subject as part of an autologous stem cell transplant procedure or an allogeneic stem cell transplant procedure.