MND PROMOTER CHIMERIC ANTIGEN RECEPTORS
The invention provides vector compositions for delivering improved adoptive T cell therapies.
1 - 56 . (canceled)
57 . A polynucleotide comprising from 5′ to 3′ a myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer-binding site substituted (MND) promoter and 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 hinge domain,
(c) a transmembrane domain 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 from a polypeptide 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 from CD8α.
59 . The polynucleotide of claim 57 , wherein the one or more co-stimulatory signaling domains is from CD28.
60 . The polynucleotide of claim 57 , wherein the one or more co-stimulatory signaling domains is from CD134.
61 . The polynucleotide of claim 57 , wherein the one or more co-stimulatory signaling domains is from CD137.
62 . The polynucleotide of claim 57 , wherein the hinge domain is a CD8a hinge domain.
63 . The polynucleotide of claim 57 , wherein the CAR comprises a signal peptide.
64 . The polynucleotide of claim 63 , wherein the signal peptide comprises an IgG1 heavy chain signal peptide, a CD8a signal peptide, or a human GM-CSF receptor alpha signal peptide.
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 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 from 5′ to 3′: a left (5′) retroviral LTR comprising a promoter; a Psi (Ψ) packaging signal; a central polypurine tract/DNA flap (cPPT/FLAP); a retroviral export element; the MND promoter; the CAR; and a right (3′) retroviral LTR.
68 . The lentiviral vector of claim 65 , 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 65 , wherein the lentiviral vector does not comprise a hepatitis B virus posttranscriptional regulatory element (HPRE) or a woodchuck post-transcriptional regulatory element (WPRE).
70 . The lentiviral vector of claim 67 , wherein:
a) the promoter of the 5′ LTR is replaced with a heterologous promoter; or
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, and a Simian Virus 40 (SV40) promoter.
71 . 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; or
d) the 3′ LTR is a self-inactivating (SIN) LTR.
72 . An isolated immune effector cell comprising the lentiviral vector of claim 65 .
73 . The immune effector cell of claim 72 , wherein the immune effector cell is a T lymphocyte.
74 . A composition comprising the immune effector cell of claim 72 and a physiologically acceptable excipient.
75 . A composition comprising the immune effector cell of claim 73 and a physiologically acceptable excipient.
76 . A method of making an immune effector cell comprising isolating CD34+ cells from bone marrow, cord blood or mobilized peripheral blood cells from a subject, and introducing the polynucleotide of claim 57 into the isolated CD34+ cells.