Compositions and methods for generating a persisting population of T cells useful for the treatment of cancer

Patent metadata

Field	Value
Publication number	US-11299536-B2
Application number	US-201816014133-A
Country	US
Kind code	B2
Filing date	Jun 21, 2018
Priority date	Feb 22, 2012
Publication date	Apr 12, 2022
Grant date	Apr 12, 2022

Abstract

The present invention provides compositions and methods for generating a genetically modified T cells comprising a chimeric antigen receptor (CAR) having an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein the T cell exhibits prolonged exponential expansion in culture that is ligand independent and independent of the addition of exogenous cytokines or feeder cells.

First claim

What is claimed is:

1. A lentiviral vector comprising a nucleic acid sequence encoding a continuous chimeric antigen receptor (CAR),
(a) wherein the CAR comprises:
(i) an anti-mesothelin antigen binding domain, a hinge domain, a transmembrane domain, a CD28 costimulatory signaling region, and a CD3 zeta signaling domain, or
(ii) an anti-c-Met antigen binding domain, an IgG4 hinge domain, a CD28 transmembrane domain, a CD28 costimulatory signaling region, and a CD3 zeta signaling domain:

(b) wherein the nucleic acid sequence encoding the CAR is operably linked to an elongation Growth Factor-1α (EF-1α) promoter, and
(c) wherein, when the lentiviral vector is transduced into a T cell, the CAR contributes to constitutive secretion of IL-2 and increased proliferation of the transduced T cell in the absence of ligand, exogenous cytokine or feeder cells as compared to an untransduced T cell.

2. The lentiviral vector of claim 1 , wherein the CAR comprises the anti-mesothelin antigen binding domain, and wherein the hinge domain is an IgG4 hinge domain or a CD8α hinge domain.

3. The lentiviral vector of claim 1 , wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

4. The lentiviral vector of claim 1 , wherein the CAR comprises the anti-mesothelin antigen binding domain, and wherein the transmembrane domain is a CD28 transmembrane domain.

5. The lentiviral vector of claim 1 , wherein the CAR comprises an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 1 or 2.

6. A T cell comprising a lentiviral vector comprising a nucleic acid sequence encoding a continuous chimeric antigen receptor (CAR), wherein:
(a) the CAR comprises:
(i) an anti-mesothelin antigen binding domain, a hinge domain, a transmembrane domain, a CD28 costimulatory signaling region, and a CD3 zeta signaling domain, or
(ii) an anti-c-Met antigen binding domain, an IgG4 hinge domain, a CD28 transmembrane domain, a CD28 costimulatory signaling region, and a CD3 zeta signaling domain;

(b) the nucleic acid sequence encoding the CAR is operably linked to an elongation Growth Factor-1α (EF-1α) promoter, and
(c) when the lentiviral vector is transduced into a T cell, the CAR contributes to constitutive secretion of IL-2 and increased proliferation of the transduced T cell in the absence of ligand, exogenous cytokine or feeder cells as compared to an untransduced T cell.

7. The T cell of claim 6 , wherein the CAR comprises the anti-mesothelin antigen binding domain, and wherein the hinge domain is an IgG4 hinge domain or a CD8α hinge domain.

8. The T cell of claim 6 , wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

9. The T cell of claim 6 , wherein the CAR comprises the anti-mesothelin antigen binding domain, and wherein the transmembrane domain is a CD28 transmembrane domain.

10. The T cell of claim 6 , wherein the CAR comprises an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 1 or 2.

11. The T cell of claim 6 , wherein the cell exhibits an anti-tumor immunity when the antigen binding domain binds to its corresponding antigen.

12. A persisting population of genetically modified T cells, wherein the T cells comprise a lentiviral vector comprising a nucleic acid sequence encoding a continuous chimeric antigen receptor (CAR), wherein:
(a) the CAR comprises:
(i) an anti-mesothelin antigen binding domain, a hinge domain, a transmembrane domain, a CD28 costimulatory signaling region, and a CD3 zeta signaling domain, or
(ii) an anti-c-Met antigen binding domain, an IgG4 hinge domain, a CD28 transmembrane domain, a CD28 costimulatory signaling region, and a CD3 zeta signaling domain;

(b) the nucleic acid sequence encoding the CAR is operably linked to an elongation Growth Factor-1α (EF-1α) promoter, and
(c) when the lentiviral vector is transduced into a T cell, the CAR contributes to constitutive secretion of IL-2 and increased proliferation of the transduced T cell in the absence of ligand, exogenous cytokine or feeder cells as compared to an untransduced T cell.

13. The persisting population of genetically modified T cells of claim 12 , wherein the genetically modified T cells exhibit an anti-tumor immunity when the antigen binding domain binds to its corresponding antigen.

14. The persisting population of genetically modified T cells of claim 12 , wherein the T cells exhibit a cytokine signature comprising at least one cytokine selected from the group consisting of IFN-γ, TNF-α, IL-1 7A, IL-2, IL-3, IL-4, GM-CSF, IL-10, IL-13, Granzyme B, Perforin, and any combination thereof.

15. The lentiviral vector of claim 1 , wherein the CAR further contributes to increased antigen-independent activation of the T cell, increased mean cell volume (MCV) of the T cell, increased cell population expansion of a population of the T cell, increased numbers of progeny of the T cell, increased effector cytokine secretion, sustained expression of granzyme, increased persistence of a population of the T cell in vitro, or increased persistence of a population of the T cell in vivo.

16. The T cell of claim 6 , wherein the CAR further contributes to increased antigen-independent activation of the T cell, increased mean cell volume (MCV) of the T cell, increased cell population expansion of a population of the T cell, increased numbers of progeny of the T cell, increased effector cytokine secretion, sustained expression of granzyme, increased persistence of a population of the T cell in vitro, or increased persistence of a population of the T cell in vivo.

17. The persisting population of genetically modified T cells of claim 12 , wherein the CAR further contributes to increased antigen-independent activation of the T cells, increased mean cell volume (MCV) of the T cells, increased cell population expansion of the T cells, increased numbers of progeny of the T cells, increased effector cytokine secretion, sustained expression of granzyme, increased persistence of the T cell population in vitro, or increased persistence of the T cell population in vivo.

18. The persisting population of genetically modified T cells of claim 12 , wherein the CAR comprises the anti-mesothelin antigen binding domain and further wherein the hinge domain is an IgG4 hinge domain or a CD8α hinge domain.

19. The persisting population of genetically modified T cells of claim 12 , wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

20. The persisting population of genetically modified T cells of claim 12 , wherein the CAR comprises the anti-mesothelin antigen binding domain and further wherein the transmembrane domain is a CD28 transmembrane domain.

21. The persisting population of genetically modified T cells of claim 12 , wherein the CAR comprises an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 1 or 2.

22. The persisting population of genetically modified T cells of claim 12 , wherein the cells exhibits an anti-tumor immunity when the antigen binding domain binds to its corresponding antigen.

Assignees

• Univ Pennsylvania

Inventors

• Frigault Matthew J
• Zhao Yangbing
• Scholler John
• June Carl H

Classifications

• C07K16/18 Primary

  against material from animals or humans · CPC title

• A61K40/31 Primary

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/4255

  Mesothelin [MSLN] · CPC title

• A61K40/4211

  CD19 or B4 · CPC title

• A61K40/4202

  Receptors, cell surface antigens or cell surface determinants · CPC title