Mnd promoter chimeric antigen receptors

Abstract

The invention provides vector compositions for delivering improved adoptive T cell therapies.

1 - 56 . (canceled) 
     
     
         57 . A polynucleotide comprising a myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer-binding site substituted (MND) promoter operably linked to a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR comprises:
 (a) an scFv that binds B cell maturation antigen (BCMA);   (b) a CD8α hinge domain;   (c) a transmembrane domain derived from a polypeptide selected from the group consisting of: CD8α; CD4, CD28, CD45, PD1, and CD152;   (d) one or more intracellular co-stimulatory signaling domains selected from the group consisting of: CD28, CD54 (ICAM), CD134 (OX40), CD137 (41BB), CD152 (CTLA4), CD273 (PD-L2), CD274 (PD-L1), and CD278 (ICOS); and   (e) a CD3ζ primary signaling domain.   
     
     
         58 . The polynucleotide of  claim 57 , wherein the transmembrane domain is derived from CD8α. 
     
     
         59 . The polynucleotide of  claim 57 , wherein the one or more co-stimulatory signaling domains is CD28. 
     
     
         60 . The polynucleotide of  claim 57 , wherein the one or more co-stimulatory signaling domains is CD134. 
     
     
         61 . The polynucleotide of  claim 57 , wherein the one or more co-stimulatory signaling domains is CD137. 
     
     
         62 . The polynucleotide of  claim 57 , further comprising a signal peptide. 
     
     
         63 . The polynucleotide of  claim 62 , wherein the signal peptide comprises an IgG1 heavy chain signal polypeptide, a CD8α signal polypeptide, or a human GM-CSF receptor alpha signal peptide. 
     
     
         64 . The polynucleotide of  claim 57 , wherein the polynucleotide encodes a CAR as set forth in any one of SEQ ID NOs: 2-3. 
     
     
         65 . A lentiviral vector comprising the polynucleotide of  claim 57 . 
     
     
         66 . The lentiviral vector of  claim 65 , wherein the lentiviral vector is selected from the group consisting essentially of human immunodeficiency virus (HIV); visna-maedi virus (VMV) virus; caprine arthritis-encephalitis virus (CAEV); equine infectious anemia virus (EIAV); feline immunodeficiency virus (FIV); bovine immune deficiency virus (BIV); and simian immunodeficiency virus (SIV). 
     
     
         67 . The lentiviral vector of  claim 65 , comprising a left (5′) retroviral LTR, a Psi (Ψ) packaging signal, a central polypurine tract/DNA flap (cPPT/FLAP), a retroviral export element; the MND promoter operably linked to the CAR of claim  1 ; and a right (3′) retroviral LTR. 
     
     
         68 . The lentiviral vector of  claim 65 , further comprising:
 a) a heterologous polyadenylation sequence;   b) a heterologous polyadenylation sequence that is a bovine growth hormone polyadenylation or signal rabbit β-globin polyadenylation sequence; or   c) a hepatitis B virus posttranscriptional regulatory element (HPRE) or woodchuck post-transcriptional regulatory element (WPRE).   
     
     
         69 . The lentiviral vector of  claim 67 , wherein:
 a) the promoter of the 5′ LTR is replaced with a heterologous promoter;   b) the promoter of the 5′ LTR is replaced with a heterologous promoter selected from the group consisting of: a cytomegalovirus (CMV) promoter, a Rous Sarcoma Virus (RSV) promoter, or a Simian Virus 40 (SV40) promoter.   
     
     
         70 . The lentiviral vector of  claim 67 , wherein:
 a) the 5′ LTR or 3′ LTR is a lentivirus LTR;   b) the 3′ LTR comprises one or more modifications;   c) the 3′ LTR comprises one or more deletions;   d) the 3′ LTR is a self-inactivating (SIN) LTR.   
     
     
         71 . An isolated immune effector cell comprising the lentiviral vector of  claim 57 . 
     
     
         72 . The immune effector cell of  claim 71 , wherein the immune effector cell is a T lymphocyte. 
     
     
         73 . A composition comprising the immune effector cell of  claim 71  and a physiologically acceptable excipient. 
     
     
         74 . A composition comprising the immune effector cell of  claim 72  and a physiologically acceptable excipient. 
     
     
         75 . A method of making an immune effector cell comprising isolating CD34+ cells from bone marrow, cord blood or mobilized peripheral blood from a subject, and introducing the polynucleotide of  claim 57  into the isolated CD34+ cells. 
     
     
         76 . A method of treating a cancer in a subject in need thereof, comprising administering to the subject a therapeutically effect amount of the composition of  claim 74 .