Human monoclonal antibodies specific for glypican-3 and use thereof
Inventors
Ho, Mitchell • Feng, Mingqian • Dimitrov, Dimiter S. • Gao, Wei
Assignees
US Department of Health and Human Services
Publication Number
US-9206257-B2
Publication Date
2015-12-08
Expiration Date
2032-04-19
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Abstract
Described herein is the identification of human monoclonal antibodies that bind GPC3 or heparan sulfate (HS) chains on GPC3 with high affinity. The antibodies described herein are capable of inhibiting HCC cell growth and migration. Provided are human monoclonal antibodies specific for GPC3 or HS chains on GPC3, including immunoglobulin molecules, such as IgG antibodies, as well as antibody fragments, such as single-domain VH antibodies or single chain variable fragments (scFv). Further provided are compositions including the antibodies that bind GPC3 or HS chains on GPC3, nucleic acid molecules encoding these antibodies, expression vectors comprising the nucleic acids, and isolated host cells that express the nucleic acids. Methods of treating cancer and/or inhibiting tumor growth or metastasis are also provided. Further provided are methods of detecting cancer in a subject and confirming a diagnosis of cancer in a subject.
Core Innovation
This invention relates to the identification and characterization of human monoclonal antibodies that bind specifically to glypican-3 (GPC3) or heparan sulfate (HS) chains on GPC3 with high affinity. These antibodies include immunoglobulin molecules such as IgG antibodies, single-domain VH antibodies, and single chain variable fragments (scFv). The invention also provides compositions including these antibodies, nucleic acid molecules encoding them, expression vectors, host cells expressing these nucleic acids, and immunoconjugates comprising the antibodies linked to effector molecules such as toxins.
The invention addresses the problem of liver cancer treatment, particularly hepatocellular carcinoma (HCC), which is prevalent and has poor response to conventional chemotherapy. Surgery is the standard treatment but often not feasible at advanced stages due to lack of effective drugs with different mechanisms of action. Immunotherapy is a promising approach but is limited by a lack of good tumor-specific targets. GPC3 is highly expressed in HCC and other cancers, but not in normal tissues, making it a suitable target. However, the precise biological functions and roles of GPC3 in tumorigenesis were previously unknown.
The invention provides methods to confirm cancer diagnosis and detect cancer by using human monoclonal antibodies that specifically bind GPC3 or its HS chains. Further, the antibodies and immunoconjugates are demonstrated to inhibit tumor growth, migration, and metastasis in vitro and in vivo, including through mechanisms such as cell cycle arrest, apoptosis, inhibition of Wnt signaling, and Yap inactivation. The antibodies can also be engineered into immunotoxins or chimeric antigen receptors for targeted cancer therapy.
Claims Coverage
The patent claims cover one independent inventive feature related to an isolated human VH single-domain monoclonal antibody that specifically binds glypican-3 (GPC3) as set forth in amino acid sequence SEQ ID NO: 2 and its various applications and compositions.
Isolated human VH single-domain monoclonal antibody binding GPC3
An isolated human variable heavy (VH) single domain monoclonal antibody that binds glypican-3 (GPC3), comprising amino acid residues 26-33, 51-57 and 96-105 of SEQ ID NO: 2, including the full sequence of SEQ ID NO: 2, with the antibody optionally labeled for detection.
Composition comprising the VH single-domain antibody
A composition comprising a therapeutically effective amount of the human VH single-domain antibody and a pharmaceutically acceptable carrier.
Immunoconjugate comprising the VH single-domain antibody and a toxin
An isolated immunoconjugate comprising the human VH single-domain antibody and an effector molecule, wherein the effector molecule is a toxin such as Pseudomonas exotoxin or variant PE38, including the amino acid sequence of SEQ ID NO: 29.
Methods of treating cancer using the VH single-domain antibody
Methods of treating a subject with cancer expressing GPC3, or inhibiting tumor growth or metastasis, by administering a therapeutically effective amount of the composition comprising the human VH single-domain antibody.
Methods of detecting or confirming cancer using the VH single-domain antibody
Methods for detecting presence of cancer or confirming diagnosis of cancer in a subject by contacting a biological sample with the human VH single-domain antibody and detecting antibody binding to the sample, where increased binding relative to a control indicates presence of cancer.
Nucleic acids, expression vectors and host cells encoding the VH single-domain antibody
Isolated nucleic acid molecules encoding the human VH single-domain monoclonal antibody, including sequences of SEQ ID NO: 1, expression vectors comprising these nucleic acids, and transformed host cells.
The claims principally cover human VH single-domain monoclonal antibodies specifically binding GPC3, their compositions including immunoconjugates with toxins, methods for cancer treatment and diagnosis employing these antibodies, and nucleic acids and host cells encoding them.
Stated Advantages
Human monoclonal antibodies are fully human and less immunogenic compared to murine antibodies.
Single-domain antibodies have better tumor penetration due to small size, easier production, and feasibility in various therapeutic formats such as bispecific antibodies and immunotoxins.
The antibodies bind with high affinity to native, cell surface-associated GPC3 and can directly inhibit HCC cell proliferation and migration.
HS20 antibody specifically inhibits cancer cell migration by disrupting the interaction between GPC3 and Wnt3a, thus inhibiting Wnt signaling.
Immunotoxins comprising the antibodies exhibit strong cytotoxic activity against GPC3-positive cancer cells and inhibit tumor growth in vivo.
Documented Applications
Confirming the diagnosis of cancers that express GPC3, including hepatocellular carcinoma (HCC), melanoma, lung cancer, and ovarian cancer, by detecting antibody binding in biological samples.
Detecting cancers expressing GPC3 in subjects by contacting samples with the antibodies and measuring antibody binding.
Treating cancers expressing GPC3, including HCC, by administering therapeutically effective amounts of the human monoclonal antibodies or immunoconjugates comprising these antibodies.
Inhibiting tumor growth and metastasis in subjects with GPC3-expressing cancers by antibody administration.
Engineering cytotoxic T lymphocytes (CTLs) expressing chimeric antigen receptors (CARs) comprising antigen-binding fragments of the human antibodies for adoptive cell therapy against GPC3-positive tumors.
Use of bispecific antibodies comprising the GPC3-specific antibodies and antibodies targeting T cell receptors (e.g., CD3) to recruit immune cells to GPC3-expressing cancers for treatment.
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