MND promoter chimeric antigen receptors
Inventors
Morgan, Richard • Friedman, Kevin • Ryu, Byoung
Assignees
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Publication Number
US-10774343-B2
Publication Date
2020-09-15
Expiration Date
Abstract
Vector compositions comprising a myeloproliferative sarcoma virus enhancer, negative control region deleted, dl587rev primer-binding site substituted (MND) promoter operably linked to a chimeric antigen receptor (CAR) are provided.
Core Innovation
The invention relates to a lentiviral vector that drives persistent expression of a chimeric antigen receptor (CAR) using a myeloproliferative sarcoma virus enhancer, negative control region deleted and dl587rev primer-binding site substituted, to form an MND promoter operably linked to a nucleic acid encoding the CAR. The CAR includes a signal peptide, an scFv that binds B cell maturation antigen (BCMA), a CD8α hinge domain, a CD8α transmembrane domain, a CD137 co-stimulatory signaling domain, and a CD3ζ primary signaling domain.
The disclosed promoter concept uses the myeloproliferative sarcoma virus enhancer with deletion of the negative control region and substitution of the dl587rev primer-binding site to provide an MND promoter that is operably linked to the CAR-encoding nucleic acid within a lentiviral vector framework. The background problem addressed is durable CAR T-cell function in cancer therapeutics.
The invention also relates to genetically modified immune effector cells, including T lymphocytes and CD34+ progenitors, for adoptive immunotherapy for cancer treatment. The document further describes storage by cryopreservation and pharmaceutical compositions comprising the CAR-modified cells with physiologically acceptable excipients or formulations, and it introduces companion diagnostic concepts tied to the same CAR-target biomarkers.
Claims Coverage
The claim set centers on an MND promoter/enhancer-driven lentiviral vector encoding a BCMA CAR with specified hinge, transmembrane, co-stimulatory, and CD3ζ signaling domains. Dependent claims further specify lentiviral regulatory components, SIN LTR modifications/deletions, named polyadenylation and post-transcriptional regulatory elements, optional selection of lentivirus type, and a CD34+ cell introduction context; at minimum, six inventive themes are captured.
MND promoter with myeloproliferative sarcoma virus enhancer configuration
A lentiviral vector comprising a polynucleotide having 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).
BCMA-binding CAR architecture with specified signaling modules
The CAR comprises a signal peptide; an scFv that binds B cell maturation antigen (BCMA); a CD8α hinge domain; a CD8α transmembrane domain; a CD137 co-stimulatory signaling domain; and a CD3ζ primary signaling domain.
Lentiviral vector regulatory elements including LTR, packaging, and export features
The lentiviral vector comprises left (5′) retroviral LTR, Psi (Ω) packaging signal, cPPT/FLAP, a retroviral export element, and a right (3′) retroviral LTR, with the MND promoter operably linked to the CAR.
Self-inactivating 3′ LTR with modifications and deletions
The lentiviral vector has a 5′ and/or 3′ lentivirus LTR where the 3′ LTR includes one or more modifications and one or more deletions, and the 3′ LTR is a self-inactivating (SIN) LTR.
Heterologous polyadenylation and post-transcriptional regulatory elements
The lentiviral vector comprises a heterologous polyadenylation sequence, including bovine growth hormone polyadenylation or rabbit β-globin polyadenylation, and/or includes HPRE or WPRE post-transcriptional regulatory elements.
Selected lentiviral vector type from named viruses
The lentiviral vector is selected from the group consisting of HIV, VMV, CAEV, EIAV, FIV, BIV, and SIV.
Immune effector cell embodiment with CD34+ cells introduction
A method of making an immune effector cell comprises isolating CD34+ cells from bone marrow, cord blood or mobilized peripheral blood from a subject and introducing the lentiviral vector into the isolated CD34+ cells.
Overall, the claims define an MND promoter/enhancer-driven lentiviral vector expressing a BCMA-targeting CAR, with dependent features covering lentiviral regulatory elements, SIN LTR structure, named polyadenylation and post-transcriptional regulatory elements, vector type selection, and a CD34+ cell embodiment.
Stated Advantages
Durable CAR T-cell function in cancer therapeutics.
Documented Applications
Genetically modified immune effector cells, including T lymphocytes and CD34+ progenitors, for adoptive immunotherapy for cancer treatment.
Storage by cryopreservation of CAR-expressing immune effector cells.
Pharmaceutical compositions comprising the CAR-modified cells with physiologically acceptable excipients or formulations.
Companion diagnostic concepts tied to the same CAR-target biomarkers.
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