Novel In Vitro and In Vivo Enrichment Strategy Targeting Lymphocytes Derived from Vector Transduced HSCS for Therapy of Disorders

Abstract

The present invention is related to a dual promoter lentiviral vector and methods of use for the treatment of diseases and disorders, specifically lysosomal storage disorders.

We claim: 
     
         1 . A dual promoter lentivirus vector that expresses a protein of interest encoded by a transgene and a mutant form of inosine-5′-monophosphate dehydrogenase 2 (IMPDH2(IY)) when transduced into a host cell. 
     
     
         2 . The dual promoter lentivirus vector of  claim 1 , wherein the vector when transduced into the host cell confers resistance to mycophenolic acid (MPA) and/or mycophenolate mofetil (MMF) in vitro and/or in vivo. 
     
     
         3 . The dual promoter lentivirus vector of  claim 1 , wherein the host cell is a hematopoietic cell. 
     
     
         4 . The dual promoter lentivirus vector of  claim 1 , wherein a first promoter is hPGK promoter and a second promoter is EFla promoter. 
     
     
         5 . The dual promoter lentivirus vector of  claim 1 , wherein the mutant form of inosine-5′-monophosphate dehydrogenase 2 (IMPDH2) contains T333I and S351Y mutations in the IMPDH2 transgene. 
     
     
         6 . The dual promoter lentivirus vector of  claim 1 , wherein the protein of interest is a protein or an enzyme associated with a lysosomal storage disorder. 
     
     
         7 - 8 . (canceled) 
     
     
         9 . The dual promoter lentivirus vector of claim  7 , wherein the vector is encoded by the DNA sequence of SEQ ID NO: 3 or a DNA sequence with 80% identity to SEQ ID NO:3. 
     
     
         10 - 11 . (canceled) 
     
     
         12 . The dual promoter lentivirus vector of claim  10 , wherein the vector is encoded by the DNA sequence of SEQ ID NO:4 or a DNA sequence with 80% identity to SEQ ID NO:4. 
     
     
         13 - 17 . (canceled) 
     
     
         18 . A host cell expressing the dual promoter lentivirus vector of  claim 1 . 
     
     
         19 . (canceled) 
     
     
         20 . A method of treating a subject having a disease or disorder associated with a defect in a single protein, the method comprising the steps of
 i) obtaining hematopoietic stem cells (HSCs) from the subject and/or HSCs from a suitable donor,   ii) transducing the HSCs in vitro or ex vivo with a dual promoter lentivirus vector that expresses a functional form of the protein of interest and a mutant form of inosine-5′-monophosphate dehydrogenase 2 (IMPDH2(IY)) encoded by SEQ ID NO:7 in the transduced cells,   iii) introducing the transduced HSCs into the subject, and   iv) administering to the subject an amount of mycophenolate mofetil (MMF) sufficient to enrich the population of lentivirus vector transduced HSCs engrafted in the subject, wherein the method treats one or more symptoms of the disease or disorder.   
     
     
         21 . The method of  claim 20 , wherein the subject has a lysosomal storage disorder (LSD), and wherein the protein of interest is associated with the LSD. 
     
     
         22 - 32 . (canceled) 
     
     
         33 . Use of the dual promoter lentivirus vector of  claim 1  for the treatment of a disease or disorder. 
     
     
         34 . The dual promoter lentivirus vector of  claim 1 , wherein the protein of interest is a protein or an enzyme associated with a hematopoietic disease. 
     
     
         35 . The dual promoter lentivirus vector of  claim 34 , wherein the hematopoietic disease is a blood clotting disorder. 
     
     
         36 . The dual promoter lentivirus vector of  claim 35 , wherein the blood clotting disorder is hemophilia. 
     
     
         37 . The dual promoter lentivirus vector of  claim 35 , wherein the transgene of interest is Factor VIII (SEQ ID NO:11) or a DNA sequence with at least 80% identity to SEQ IDNO:11. 
     
     
         38 . The method of  claim 20 , wherein the subject has a hematopoietic disease, and wherein the protein of interest is associated with the hematopoietic disease. 
     
     
         39 . The method of  claim 38 , wherein the hematopoietic disease is a blood clotting disease. 
     
     
         40 . The method of  claim 39 , wherein the blood clotting disease is hemophilia and the protein of interest is Factor VIII encoded by the transgene of SEQ ID NO:11 or a DNA sequence with at least 8% T identity to SEQ ID NO:11.