Methods and compositions for engineering CD4-deficient CAR T cells and anti-CD4 CAR T cells and uses thereof
AssigneeSeattle Children's Hospital
InventorsThor Wagner, Jaya Sahni
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Abstract
Some aspects of the methods and compositions provided herein relate to the disruption of at least one CD4 gene in a cell, such as a CD4+ T cell. In some embodiments, the disruption comprises use of a CRISPR guide polynucleotide. Some embodiments also include the preparation and use of a cell having at least one disrupted CD4 gene and a chimeric antigen receptor (CAR). Some aspects of the methods and compositions provided herein relate to CARs, such as an anti-CD4 CAR or an anti-CD19 CAR, and use to treat disorders including HIV, acute myeloid leukemia (AML), and acute lymphocytic leukemia (ALL).
Claims20
A genetically modified cell comprising an inactivated endogenous CD4 gene, wherein the genetically modified cell:
- comprises a chimeric antigen receptor (CAR), wherein the CAR specifically binds a target antigen;
- and
- wherein the genetically modified cell is prepared by introducing into a source cell a guide RNA (gRNA), which hybridizes to a first exon or a third exon of a CD4 gene, wherein the source cell is a CD3+ cell;
- and
- wherein the genetically modified cell comprises an activity to produce a cytokine against a target cell comprising the target antigen, wherein a level of the cytokine produced by the genetically modified cell is the same or greater than a level of the cytokine produced by a cell comprising the CAR and which lacks an inactivated endogenous CD4 gene.
A CD3+ cell comprising an inactivated endogenous CD4 gene, the cell comprising:
- a guide RNA (gRNA), which hybridizes to a first exon or a third exon of a CD4 gene,
- a Cas 9 nuclease or polynucleotide encoding the Cas 9 nuclease, and
- a polynucleotide encoding a chimeric antigen receptor (CAR), wherein the CAR specifically binds a target antigen;
- and
- wherein the cell comprises an activity to produce a cytokine against a target cell comprising the target antigen, wherein a level of the cytokine produced by the cell is the same or greater than a level of the cytokine produced by a cell comprising the CAR and which lacks an inactivated endogenous CD4 gene.
Dependencies are inferred from the claim text.