Antigen-binding proteins targeting CD56 and uses thereof
Inventors
Sadelain, Michel • Benjamin, Reuben • Dimitrov, Dimiter S. • Feng, Yang
Assignees
Memorial Sloan Kettering Cancer Center • US Department of Health and Human Services
Publication Number
US-10730941-B2
Publication Date
2020-08-04
Expiration Date
2036-08-01
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Abstract
The presently disclosed subject matter provides for methods and compositions for treating cancer (e.g., multiple myeloma). It relates to anti-CD56 antibodies, chimeric antigen receptors (CARs) that specifically target human CD56, and immunoresponsive cells comprising such CARs. The presently disclosed CD56-specific CARs have enhanced immune-activating properties, including anti-tumor activity.
Core Innovation
The presently disclosed subject matter provides methods and compositions for treating cancer, such as multiple myeloma, using antigen-binding proteins including antibodies or antigen-binding portions thereof, and chimeric antigen receptors (CARs) that specifically target human CD56. It further includes immunoresponsive cells comprising such CARs and methods of using these for treating cancers. The CD56-specific CARs described have enhanced immune-activating properties, including anti-tumor activity.
The background identifies multiple myeloma as the second most common hematological malignancy, remaining incurable despite advances with immunomodulatory drugs and proteasome inhibitors. Various immunotherapeutic strategies are under investigation to improve disease-free survival. CD56 is identified as a frequently expressed antigen in myeloma, playing roles in tumorigenesis by mediating adhesion, migration, invasion, proliferation, and inhibiting apoptosis. However, CD56 is also expressed on normal tissues, raising concerns of off-tumor toxicity in CAR therapies. There is a need for novel CAR designs targeting antigens like CD56 highly expressed on myeloma cells with limited normal tissue expression to induce potent tumor eradication with minimal toxicity.
The summary details CARs comprising extracellular antigen-binding domains that cross-compete with or bind the same epitope as reference antibodies targeting human CD56, specifying particular complementarity determining region (CDR) sequences. The disclosed CARs exhibit specific binding affinities to human CD56, generally around or below 3×10−9 M. Immunoresponsive cells, such as T cells or NK cells, expressing these CARs exhibit antigen-dependent activation, proliferation, cytokine secretion, and cytotoxicity against CD56-expressing tumor cells. The CARs can include defined transmembrane and intracellular domains, including CD28 and CD3ζ polypeptides, to optimize signaling. The invention also contemplates use of these CAR-expressing cells and anti-CD56 antibodies to reduce tumor burden and increase survival in subjects with cancers associated with CD56 overexpression, including multiple myeloma.
Claims Coverage
The claims include multiple inventive features focusing on CARs that specifically bind human CD56 through defined heavy and light chain complementarity determining regions, their structural components, and immunoresponsive cells comprising them.
Chimeric antigen receptor with defined CD56 binding CDRs
A CAR comprising an extracellular antigen-binding domain binding human CD56, with heavy chain variable region CDRs comprising amino acid sequences in SEQ ID NOs: 1, 2, and 3 or 59, and light chain variable region CDRs comprising amino acid sequences in SEQ ID NOs: 4, 5, and 6 or variants thereof. The CAR includes a transmembrane domain and an intracellular domain.
CARs with specific combinations of variable region sequences
CARs wherein the heavy and light chain variable regions comprise amino acid sequences selected from specific SEQ ID NO pairs including (7 and 8), (19 and 20), (21 and 22), (23 and 24), or (25 and 26), defining particular human scFvs targeting CD56.
Extracellular antigen-binding domain formats
CARs wherein the extracellular antigen-binding domain is a single-chain variable fragment (scFv), a Fab optionally crosslinked, or a F(ab)2, including human scFvs or fusion proteins that comprise these domains.
Transmembrane and intracellular domain composition
CARs wherein the transmembrane domain comprises one or more polypeptides selected from CD8, CD28, CD3ζ, CD4, 4-1BB, OX40, ICOS, CTLA-4, PD-1, LAG-3, 2B4, BTLA, or combinations thereof. The intracellular domain includes a CD3ζ polypeptide and optionally at least one co-stimulatory signaling domain such as CD28, 4-1BB, OX40, ICOS, or DAP-10.
Immunoresponsive cells expressing CARs
Isolated immunoresponsive cells, including T cells, NK cells, stem cells, or progenitor cells, constitutively expressing the disclosed CARs on their surface and optionally co-expressing an antigen recognizing receptor for a second antigen distinct from CD56.
Nucleic acid molecules and vectors encoding CARs
Nucleic acid molecules encoding the CARs and vectors comprising these nucleic acids for expression in host cells to produce CAR-expressing immunoresponsive cells.
Methods of producing and using CAR-expressing immunoresponsive cells
Methods of producing immunoresponsive cells binding human CD56 by introducing nucleic acid sequences encoding the CAR, methods of reducing tumor burden and increasing survival in subjects by administering CAR-expressing cells, pharmaceutical compositions containing the immunoresponsive cells, and kits comprising the cells and instructions for use.
The claims cover CARs with carefully defined anti-CD56 antigen-binding domains, their structural domains, nucleic acid vectors encoding them, immunoresponsive cells expressing these CARs, methods for producing and using these cells for treating CD56-expressing tumors, pharmaceutical compositions, and treatment kits. The features focus on the antigen-binding specificity and affinity, structural elements of CARs, and therapeutic applications.
Stated Advantages
The CARs demonstrate striking activity against high CD56-expressing tumor cells while sparing low CD56-expressing normal cells such as NK and NKT cells.
Human scFv-based CARs reduce immunogenicity compared to murine-derived CARs.
CD56-targeted CAR T cells efficiently lyse myeloma cell lines and primary myeloma cells in vitro, and eradicate myeloma tumor cells and prevent bone disease in vivo in a xenograft model.
Optimization of CAR affinity and epitope selection allows discrimination between high antigen density tumor cells and low antigen density normal cells, potentially reducing off-tumor toxicity.
Documented Applications
Treatment of cancers, specifically multiple myeloma, neuroblastoma, glioma, acute myeloid leukemia, colon cancer, pancreatic cancer, thyroid cancer, small cell lung cancer, and NK cell lymphoma.
Use of anti-CD56 antibodies, antigen-binding portions thereof, CARs, and immunoresponsive cells expressing such CARs for reducing tumor burden, inducing tumor cell death, reducing tumor size, and eradicating tumors in subjects.
Methods for increasing or lengthening survival in subjects having neoplasia through administration of the immunoresponsive cells or anti-CD56 antibodies.
Production of immunoresponsive cells expressing CARs targeting human CD56 by introducing nucleic acid sequences encoding the CARs.
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