US-12624086-B2 - Materials and methods for treating cancer

US12624086B2US 12624086 B2US12624086 B2US 12624086B2US-12624086-B2

Abstract

This document provides methods and materials involved in treating cancer. For example, chimeric antigen receptor T cells having reduced levels of GM-CSF are provided. Also provided as methods for making and using chimeric antigen receptor T cells having reduced levels of GM-CSF.

Inventors

Cameron DURRANT
Dale Chappell
SAAD J. KENDERIAN
ROSALIE M. STERNER
MICHELLE J. COX
REONA SAKEMURA

Assignees

MAYO FOUNDATION FOR MEDICAL EDUCATION AND RESEARCH

Dates

Publication Date: 20260512
Application Date: 20191031

Claims (11)

1 . A method for enhancing anti-tumor efficacy of chimeric antigen receptor-expressing T19-cell (CAR T19-cell) therapy in a subject having cancer, the method comprising administering to the subject CAR-T19 cells having a granulocyte-macrophage colony-stimulating factor (GM-CSF) gene knockout (GM-CSF k/o CAR-T19 cells), wherein administration of the GM-CSF k/o CAR-T19 cells reduces tumor burden compared to tumor burden reduction by administration of CAR-T19 cells lacking said GM-CSF gene knockout.
2 . The method of claim 1 , further comprising administering to the subject an anti-hGM-CSF antibody, wherein the anti-hGM-CSF antibody is a recombinant anti-hGM-CSF antibody that binds to and neutralizes human GM-CSF.
3 . The method of claim 2 , wherein the administration of (i) the GM-CSF k/o CAR-T19 cells or (ii) the GM-CSF k/o CAR-T19 cells and the anti-hGM-CSF antibody decreases or prevents CD14 + myeloid cell trafficking to a central nervous system (CNS) of the subject, wherein a high level of CD14 + myeloid cells in the CNS of the subject is indicative of neurotoxicity.
4 . The method of claim 2 , wherein an objective response rate of the subject administered the anti-hGM-CSF antibody is improved compared to a subject that is not administered the anti-hGM-CSF antibody.
5 . The method of claim 4 , wherein the objective response rate is a complete response rate or a partial response rate.
6 . The method of claim 2 , wherein a progression free response and/or survival of the subject is improved compared to a subject that is not administered the anti-hGM-CSF antibody.
7 . The method of claim 6 , wherein the survival is overall survival of the subject.
8 . The method of claim 2 , wherein the anti-hGM-CSF antibody is administered to the subject after administration of the GM-CSF k/o CAR-T19 cells.
9 . The method of claim 1 , wherein the cancer is lymphoma or a leukemia.
10 . The method of claim 9 , wherein the lymphoma is a diffuse large B cell lymphoma (DLBCL).
11 . The method of claim 9 , wherein the leukemia is acute lymphoblastic leukemia (ALL).

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a U.S. national stage application under 37 U.S.C. 371 of PCT International Application PCT/US2019/059275, filed Oct. 31, 2019, which claims benefit of U.S. Provisional Application No. 62/753,485 filed Oct. 31, 2018, the priority dates of which are hereby claimed, and the contents of each of which are hereby incorporated by reference in their entireties. SEQUENCE LISTING INCORPORATION The “.txt” Sequence Listing filed with this application by EFS and which is entitled P-588784-US1-SEQ-LISTING-8MAR.2022_ST25.txt, is 12.0 kilobytes in size and which was created on Mar. 8, 2022, is hereby incorporated by reference. 1. TECHNICAL FIELD This document relates to methods and materials involved in treating cancer. For example, this document provides methods and materials for using chimeric antigen receptor T cells having reduced expression levels of one or more cytokines (e.g., GM-CSF) in an adoptive cell therapy (e.g., a chimeric antigen receptor T cell therapy) to treat a mammal (e.g., a human) having cancer. 2. BACKGROUND INFORMATION Unprecedented results from pivotal trials evaluating the safety and efficacy of CD19 directed chimeric antigen receptor T cells (CART19) have led to the recent FDA approval of CART19 (Tisagenlecleucel) for relapsed refractory acute lymphoblastic leukemia (ALL) and CART19 (Axi-Cel) for the treatment of diffuse large B cell lymphoma (DLBCL). The application of CART cell therapy is associated with toxicities resulting in cytokine release syndrome (CRS) and neurotoxicity. Additionally, the efficacy of CART cell therapy is limited to only 40% durable remissions in lymphoma and 50-60% durable remissions in acute leukemia. SUMMARY This document provides methods and materials for generating T cells (e.g., chimeric antigen receptor (CAR) T cells (CARTs)) having a reduced expression level of one or more cytokine (e.g., GM-CSF) polypeptides. For example, a T cell (e.g., a CART) can be engineered to have reduced GM-CSF polypeptide expression (e.g., for use in adoptive cell therapy). In some cases, a T cell (e.g., a CART) can be engineered to knock out (KO) a nucleic acid encoding one or more cytokine polypeptides (e.g., a GM-CSF polypeptide) to reduce cytokine polypeptide (e.g., GM-CSF polypeptide) expression in that T cell. This document also provides methods and materials for using T cells (e.g., CARTs) having a reduced expression level of one or more cytokines (e.g., GM-CSF polypeptides). For example, T cells (e.g., CARTs) having a reduced level of GM-CSF polypeptides can be administered (e.g., in an adoptive cell therapy) to a mammal having cancer to treat the mammal. In one aspect, this invention provides a method for enhancing anti-tumor efficacy of immunotherapy in a subject, the method comprising administering to the subject CAR-T cells having a GM-CSF gene inactivation, GM-CSF gene knock-down or gene knockout (GM-CSFk/o CAR-T cells), wherein administration of the CAR-T cells reduces or prevents immunotherapy-related toxicity. In another aspect, this invention provides a method for reducing a level of a non-GM-CSF cytokine in a subject treated with immunotherapy, the method comprising administering to the subject CAR-T cells having a GM-CSF gene inactivation, GM-CSF gene knock-down or gene knockout (GM-CSFk/o CAR-T cells). In one aspect, this invention provides a method for GM-CSF gene inactivation, GM-CSF gene knock-down or GM-CSF knockout (KO) in a cell comprising targeted genome editing or GM-CSF gene silencing. In one aspect, this invention provides a method for making a chimeric antigen receptor T cell having a reduced level of granulocyte-macrophage colony-stimulating factor (GM-CSF) polypeptides, said method comprising: introducing a nucleic acid construct into an ex vivo T cell, wherein said nucleic acid construct comprises: a) a nucleic acid encoding a guide RNA, wherein said guide RNA is complementary to a GM-CSF messenger RNA; b) a nucleic acid encoding a Cas nuclease, and c) a nucleic acid encoding said chimeric antigen receptor. In another aspect, this invention provides a method for making a chimeric antigen receptor T cell having a reduced level of granulocyte-macrophage colony-stimulating factor (GM-CSF) polypeptides, said method comprising: introducing a complex into an ex vivo T cell, wherein said complex comprises: a) a guide RNA, wherein said guide RNA is complementary to a GM-CSF messenger RNA; and b) a Cas nuclease; and introducing a nucleic acid encoding said chimeric antigen receptor into said ex vivo T cell. In an aspect, this invention provides a method for making a chimeric antigen receptor T cell having a reduced level of cytokine polypeptides, said method comprising: introducing a nucleic acid construct into an ex vivo T cell, wherein said nucleic acid construct comprises: a) a nucleic acid encoding a guide RNA, wherein said guide RNA is complementary to a cytokine messenger RNA; b) a nucleic acid enc