US-12617849-B2 - Antigen specific CD19-targeted CAR-T cells

Abstract

Disclosed are compositions and methods for targeted treatment of cancer, such as hematologic cancer. In particular, chimeric antigen receptor (CAR) T cells are disclosed that can be used with adoptive cell transfer to target and kill cancer cells with reduced antigen escape. Therefore, also disclosed are methods of providing an anti-tumor immunity in a subject with hematologic cancer that involves adoptive transfer of the disclosed CAR T cells.

Inventors

• Blake T. Aftab

Assignees

• Atara Biotherapeutics, Inc.

Dates

Publication Date

20260505

Application Date

20200429

Claims (9)

1. 1 . An immune cell expressing a chimeric antigen receptor (CAR) polypeptide comprising a CD19-binding domain comprising the amino acid sequence set forth in SEQ ID NO:9, a hinge region comprising the amino acid sequence set forth in SEQ ID NO:12, a transmembrane domain comprising the amino acid sequence set forth in SEQ ID NO:13, an intracellular domain comprising the amino acid sequence set forth in SEQ ID NO: 14, and an intracellular signaling domain comprising the amino acid sequence set forth in SEQ ID NO: 11; wherein the immune cell is an EBV-antigen sensitized cytotoxic T lymphocyte (CTL); and wherein the CAR-expressing immune cell is not derived from the subject to whom the cell is to be administered.
2. 2 . A method of treating a B-lymphocyte antigen-associated cancer in a subject, the method comprising administering to the subject an effective amount of an adoptive immunotherapy composition comprising the CAR-expressing immune cell of claim 1 .
3. 3 . The method of claim 2 , wherein the B-lymphocyte antigen-associated cancer is an EBV-associated lymphoproliferative disease.
4. 4 . The immune cell of claim 1 , wherein the CAR polypeptide comprises at least one co-stimulatory signaling region, wherein the co-stimulatory signaling region comprises a signaling domain of any one of the polypeptides CD8, CD3ζ, CD3δ, CD3γ, CD3ε, FcγRI-γ, FcγRIII-γ, FcεRIβ, FcεRIγ, DAP10, DAP12, CD32, CD79a, CD79b, CD28, CD3C, CD4, b2c, CD137 (41BB), ICOS, CD27, CD288, CD80, NKp30, OX40, mutants thereof, or any combination thereof.
5. 5 . The immune cell of claim 1 , wherein the CAR polypeptide comprises a signal peptide comprising the amino acid sequence set forth in SEQ ID NO:8.
6. 6 . The immune cell of claim 1 , wherein the CAR polypeptide comprises the amino acid sequence set forth in SEQ ID NO:6 or SEQ ID NO:7.
7. 7 . The immune cell of claim 1 , wherein the immune cell is derived from a donor sample, or from a bank or library of donor samples.
8. 8 . The method of claim 2 , wherein the B-lymphocyte antigen-associated cancer is a hematologic cancer selected from: acute leukemia, chronic leukemia, lymphocytic leukemia, myelogenous leukemia, a pre-leukemic condition, Hodgkin lymphoma, Non-Hodgkin lymphoma, an EBV-associated lymphoproliferative disease, mature B cell neoplasm, mature T cell or natural killer (NK) cell neoplasm, precursor lymphoid neoplasm, and immunodeficiency-associated lymphoproliferative disorder.
9. 9 . The method of claim 2 , wherein the subject is human.

Description

RELATED APPLICATIONS This application is a national stage filing under 35 U.S.C. § 371 of PCT/US20/30435, filed Apr. 29, 2020, which claims the benefit of priority to U.S. Provisional Application Ser. No. 62/840,774, filed Apr. 30, 2019, which is incorporated by reference in its entirety. SEQUENCE LISTING The instant application contains a Sequence Listing, which has been filed electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Oct. 29, 2021, is named ABH-00701_SL.txt and is 20,054 bytes in size. BACKGROUND Hematologic malignancies represent some of the most common cancers occurring in both children and adults. For example, approximately 4,000 de novo cases of the aggressive B-cell lineage malignancy B-cell acute lymphoblastic leukemia (B-ALL) are diagnosed each year in the United States and represents the most common malignancy of childhood. Genetic mutations that induce aberrant arrest of normal lymphoid maturation, evasion of apoptosis and uncontrolled cellular proliferation result in over-production of B-cell lymphoblasts. In adults, over 6000 incident cases of acute lymphoblastic leukemia (ALL) occur each year. (Hanahan, D. and Weinberg, R. (2000) The hallmarks of cancer. Cell 100: 57-70; Teitell, M. and Pandolfi, P. (2009) Molecular genetics of acute lymphoblastic leukemia. Annu Rev Pathol 4: 175-198) Moreover, lymphomas (e.g., neoplasms of the lymphatic tissues) account for approximately 5% of all cases of cancer in the United States. The main classes are malignant neoplasms (that is, cancers) of the lymphocytes, a cell type present in both the lymph and the blood. In this way, lymphomas and leukemias are both malignancies (e.g., tumors) of the hematopoietic and lymphoid tissues. As lymphoproliferative disorders, lymphomas and lymphoid leukemias are closely related, to the point that some of them are called by either name (for example adult T-cell leukemia/lymphoma). Preclinical and clinical research efforts have been focused on investigating immunotherapeutic modalities that include antibody-based and/or adoptive cellular therapeutics for high-risk cancers. Such strategies rely on tumor-associated antigens so as to enable specific targeting of cancerous cells and sparing non-cancerous cells (i.e., on target/off tumor or bystander effects). Particular interest has been paid to adoptive immunotherapy approaches that involve the genetic engineering of human immune effector cells with synthetic chimeric antigen receptors (CARs) that target tumor-associated antigens expressed on the cell surface in a major histocompatibility complex (MHC) antigen-independent manner. Without being bound to any particular theory, engagement of the CAR expressed on a T cell with a cancer-associated antigen results in intracellular signaling via T cell costimulatory domains, and subsequent expansion of the CAR T cells, to induce further cancer cell killing. However, despite positive results from early-phase trials, infusion of CD19 CAR T cells into patients still results in a number of ‘on target/on tumor’ and ‘on target/off tumor’ side effects of varying severity, such as Tumor lysis syndrome (TLS), cytokine release syndrome (CRS) and macrophage activation syndrome, CNS trafficking, prolonged B-cell aplasia, and immune escape. Therefore, in view of the long-felt and unmet need described herein, improved therapies for hematologic malignancies are needed. SUMMARY The present invention is based, at least in part, on the discovery that B-lymphocyte antigens such as CD19 (B-lymphocyte antigen CD19) can be used for the targeted treatment of blood cancers (i.e. cancers of the hematopoietic and lymphoid tissues). In some aspects, provided herein are immune cells that express a chimeric antigen receptor (CAR) polypeptide that targets B lineage cells and cancer cells that arise therefrom. In some embodiments, the CARs disclosed herein comprise a B-lymphocyte antigen-targeting domain such as a CD19, CD20, and/or CD22-binding domain, a transmembrane domain, and an intracellular signaling domain. In certain preferred embodiments, the B-lymphocyte antigen-binding domain targets a wildtype and/or mutant CD19 antigen. In certain aspects, provided herein are bi-specific chimeric antigen receptor (CAR) T cells, said cells expressing a CAR polypeptide comprising a targeting domain that selectively binds a B-lymphocyte antigen (e.g., a CD19, CD20, and/or CD22 antigen associated with a hematologic malignancy such as leukemia and/or lymphoma) and a CAR polypeptide comprising a targeting domain that selectively binds to another different tumor-associated antigen. In some such embodiments, the targeting domain of the chimeric antigen receptor (e.g., a CD19 antigen-binding domain and/or the other different tumor-associated antigen-binding domain) comprise a functional antibody fragment. Preferably, the antigen-binding domain of the chimeric antigen receptors comprise a single-chain va