Constructs for chimeric antigen receptors
Inventors
KLICHINSKY, Michael • Minutolo, Nicholas G. • Anderson, Nicholas R.
Assignees
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Publication Number
US-11312939-B2
Publication Date
2022-04-26
Expiration Date
Abstract
The present disclosure pertains to immune cells comprising chimeric antigen receptors (CARs) and methods of using immune cells comprising CARs.
Core Innovation
The invention relates to methods of modifying a macrophage or monocyte by delivering a viral vector that comprises one or more nucleic acid sequences encoding a chimeric antigen receptor (CAR). The CAR includes an extracellular domain, a transmembrane domain, and an intracellular domain. The method further includes delivering at least one Vpx protein or delivering at least one Vpx mRNA prior to transduction with the viral vector, thereby producing a modified macrophage or monocyte with increased CAR expression relative to a macrophage or monocyte receiving CAR without at least one Vpx protein.
The modified macrophage or monocyte does not exhibit an M1 phenotype relative to a macrophage or monocyte not delivered at least one Vpx protein. The disclosed approach uses Vpx in combination with CAR-encoding viral vectors to increase CAR expression while retaining plasticity for M1/M2 polarization.
The disclosed content further includes immune modulation elements, including co-expression with cytokines and stimulatory ligands, and indicates functional outcomes including improved anti-tumor killing and polarization effects associated with modifying innate immune pathways.
Claims Coverage
Two independent claims are identified. Across these claims, the inventive subject matter centers on modifying macrophages or monocytes with a CAR-encoding viral vector in combination with Vpx protein or Vpx mRNA to increase CAR expression, with one claim also requiring absence of an M1 phenotype.
Viral car delivery with at least one vpx protein while avoiding m1 phenotype
Delivering to a macrophage or monocyte a viral vector comprising one or more nucleic acid sequences encoding a CAR having an extracellular domain, a transmembrane domain, and an intracellular domain, and at least one Vpx protein, to produce a modified macrophage or monocyte exhibiting increased CAR expression relative to a macrophage or monocyte comprising a CAR delivered without at least one Vpx protein, wherein the modified macrophage or monocyte does not exhibit an M1 phenotype relative to a macrophage or monocyte not delivered at least one Vpx protein.
Vpx mrna delivered prior to viral car transduction to increase car expression
Delivering to a macrophage or monocyte a viral vector comprising one or more nucleic acid sequences encoding a CAR having an extracellular domain, a transmembrane domain, and an intracellular domain, wherein the macrophage or monocyte is one or both of electroporated or transfected with at least one Vpx mRNA prior to transduction with the viral vector, thereby producing a modified macrophage or monocyte exhibiting increased CAR expression relative to a macrophage or monocyte comprising a CAR delivered by a viral vector and not delivered at least one Vpx protein.
The independent claims cover CARs encoded by a viral vector with extracellular, transmembrane, and intracellular domains, together with Vpx protein or Vpx mRNA, to increase CAR expression in macrophages or monocytes. One claim also requires that the modified cells do not exhibit an M1 phenotype.
Stated Advantages
Increased CAR expression relative to CAR delivered without at least one Vpx protein.
The modified macrophage or monocyte does not exhibit an M1 phenotype relative to a macrophage or monocyte not delivered at least one Vpx protein.
Retaining plasticity for M1/M2 polarization.
Improved anti-tumor killing.
Documented Applications
Immune-cell modification of macrophages or monocytes for CAR expression using viral delivery that includes Vpx, including implementations using Vpx mRNA prior to viral transduction.
Co-expression with cytokines and stimulatory ligands.
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