Immune cells engineered to express CD19-directed chimeric antigen receptors and uses thereof in immunotherapy
AssigneeNkarta, Inc.
InventorsJames Barnaby Trager, Luxuan Guo Buren, Chao Guo, Mira Tohmé, Ivan Chan, Alexandra Leida Liana Lazetic
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Abstract
Provided for herein in several embodiments are immune cell-based (e.g., natural killer (NK) cell) compositions comprising CD19-directed chimeric antigen receptors. In some, embodiments the anti-CD19 binder portion of the CAR is humanized. In several embodiments, the humanized anti-CD19 CAR expressing cells exhibit enhanced expression of the CAR as well as enhanced cytotoxicity and/or persistence. Several embodiments include methods of using of the anti-CD19 CAR expressing immune cells in immunotherapy.
Claims16
An immune cell that expresses a CD19-directed chimeric antigen receptor (CAR), the chimeric antigen receptor comprising:
- an extracellular anti-CD19 binding moiety comprising a single chain Fragment variable (scFv), wherein the scFv comprises:
- a variable heavy (VH) domain, wherein the VH domain comprises the amino acid sequence of SEQ ID NO:
- 120, and
- a variable light (VL) domain;
- a hinge, wherein the hinge is a CD8 alpha hinge;
- a transmembrane domain, wherein the transmembrane domain comprises a CD8 alpha transmembrane domain;
- and
- an intracellular signaling domain, wherein the intracellular signaling domain comprises:
- an OX40 subdomain, wherein the OX40 subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO. 5,
- a CD3 zeta subdomain, wherein the CD3 zeta subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO. 7, and
- wherein the immune cell also expresses membrane-bound interleukin-15 (mbIL15), wherein the mbIL15 comprises a sequence having at least 95% sequence identity to SEQ ID NO. 12.
An immune cell that expresses a CD19-directed chimeric antigen receptor, the chimeric antigen receptor comprising:
- an extracellular anti-CD19 binding moiety comprising a single chain Fragment variable (scFv), wherein the scFv comprises:
- a variable heavy (VH) domain,
- wherein the VH domain comprises the amino acid sequence of SEQ ID NO:
- 120;
- and
- a variable light (VL) domain,
- wherein the VL domain comprises the amino acid sequence of SEQ ID NO:
- 118;
- a hinge;
- a transmembrane domain;
- and
- an intracellular signaling domain, wherein the intracellular signaling domain comprises:
- an OX40 subdomain,
- a CD3 zeta subdomain;
- and
- wherein the immune cell also expresses membrane-bound interleukin-15 (mbIL15), wherein the mbIL15 comprises a sequence having at least 95% sequence identity to SEQ ID NO. 12.
An immune cell that expresses a CD19-directed chimeric antigen receptor (CAR), the chimeric antigen receptor comprising:
- an extracellular anti-CD19 binding moiety comprising a single chain Fragment variable (scFv), wherein the scFv comprises:
- a variable heavy (VH) domain, and
- a variable light (VL) domain, wherein the VL domain comprises the amino acid sequence of SEQ ID NO:
- 118;
- a hinge, wherein the hinge is a CD8 alpha hinge;
- a transmembrane domain, wherein the transmembrane domain comprises a CD8 alpha transmembrane domain;
- and
- an intracellular signaling domain, wherein the intracellular signaling domain comprises:
- an OX40 subdomain, wherein the OX40 subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO. 5,
- a CD3 zeta subdomain, wherein the CD3 zeta subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO. 7, and
- wherein the immune cell also expresses membrane-bound interleukin-15 (mbIL15), wherein the mbIL15 comprises a sequence having at least 95% sequence identity to SEQ ID NO. 12.
An immune cell that expresses a CD19-directed chimeric antigen receptor (CAR), the chimeric antigen receptor comprising:
- an extracellular anti-CD19 binding moiety comprising a single chain Fragment variable (scFv), wherein the scFv comprises:
- a variable heavy (VH) domain, and
- a variable light (VL) domain;
- a hinge, wherein the hinge is a CD8 alpha hinge;
- a transmembrane domain, wherein the transmembrane domain comprises a CD8 alpha transmembrane domain;
- and
- an intracellular signaling domain, wherein the intracellular signaling domain comprises:
- an OX40 subdomain, wherein the OX40 subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO. 5,
- a CD3 zeta subdomain, wherein the CD3 zeta subdomain is encoded by a sequence having at least 95% sequence identity to SEQ ID NO. 7, and
- wherein the immune cell also expresses membrane-bound interleukin-15 (mbIL15), wherein the mbIL15 comprises a sequence having at least 95% sequence identity to SEQ ID NO. 12, and
- wherein the CD19-directed CAR and mbIL15 are encoded by the amino acid sequence set forth in SEQ ID NO:
- 187.
An immune cell that expresses a CD19-directed chimeric antigen receptor, the chimeric antigen receptor comprising:
- an extracellular anti-CD19 binding moiety comprising a single chain Fragment variable (scFv), wherein the scFv comprises:
- a variable heavy (VH) domain,
- wherein the VH domain comprises the amino acid sequence of SEQ ID NO:
- 120;
- and
- a variable light (VL) domain,
- wherein the VL domain comprises the amino acid sequence of SEQ ID NO:
- 118;
- a hinge;
- a transmembrane domain;
- and
- an intracellular signaling domain, wherein the intracellular signaling domain comprises:
- an OX40 subdomain,
- a CD3 zeta subdomain;
- and
- wherein the immune cell also expresses membrane-bound interleukin-15 (mbIL15), wherein the mbIL15 comprises a sequence having at least 95% sequence identity to SEQ ID NO. 12,
- wherein the CD19-directed CAR and mbIL15 are encoded by the amino acid sequence set forth in SEQ ID NO:
- 187.
Dependencies are inferred from the claim text.