Compositions and methods for retrieving tumor-related antibodies and antigens
Inventors
Hua, Xianxin • He, Xin • FENG, Zijie • Siegel, Donald L.
Assignees
University of Pennsylvania Penn
Publication Number
US-12410234-B2
Publication Date
2025-09-09
Expiration Date
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Abstract
The present invention includes compositions and methods for retrieving tumor-related antibodies and antigens. In one aspect, the invention includes a method for Sequential Tumor-related Antibody and antigen Retrieving (STAR) which directly and efficiently identifies potent antibodies that can specifically bind to tumor-related antigens on the tumor cell surface. In another aspect, the invention includes a CAR comprising a nanobody, a transmembrane domain, and an intracellular domain, wherein the nanobody is retrieved by a STAR method. In another aspect, the invention includes a CAR T system that targets CD13 and treats acute myeloid leukemia. In another aspect, the invention includes a CAR T system and ADC that targets CDH17 and treats NETs and other types of tumors expressing this antigen, with tolerable toxicities.
Core Innovation
The present invention includes compositions and methods for retrieving tumor-related antibodies and antigens. In one aspect, the invention includes a method for Sequential Tumor-related Antibody and antigen Retrieving (STAR) which directly and efficiently identifies potent antibodies that can specifically bind to tumor-related antigens on the tumor cell surface. In another aspect, the invention includes a chimeric antigen receptor (CAR) comprising a nanobody, a transmembrane domain, and an intracellular signaling domain, wherein the nanobody is retrieved by a STAR method. The invention further discloses a CAR T system that targets CD13 to treat acute myeloid leukemia (AML) and a CAR T system and ADC that targets CDH17 to treat NETs and other types of tumors expressing this antigen, with tolerable toxicities.
The invention includes methods and compositions for generating and using tumor-specific and CAR T cell-compatible nanobodies and related constructs. The patent describes a method for generating a plurality of tumor-specific and CAR T cell-compatable nanobodies involving immunization of a camelid, phage display, selection of tumor specific nanobodies, insertion into CAR expressing vectors, transduction into human primary T cells, and in vivo selection of nanobodies that cause T cell enrichment in the tumor in vivo; [procedural detail omitted for safety]. In various embodiments the nanobody specifically binds to CD13 or CDH17 (including binding the first domain of CDH17), the nanobody sequences are provided (e.g., SEQ ID NOs: 1-5, 19, 23, 25-30), and the nanobodies can be used in CARs, switchable CAR systems, modified T cells, or ADCs comprising a nanobody conjugated to a drug, toxin, or radioisotope.
Claims Coverage
Two independent claims were identified and four main inventive features are extracted.
Chimeric antigen receptor comprising a nanobody that binds the first domain of CDH17
A chimeric antigen receptor (CAR) comprising a nanobody, a transmembrane domain, and an intracellular signaling domain, wherein the nanobody specifically binds to the first domain of CDH17.
Nanobody defined by specific CDR sequences
The nanobody comprises a CDR1 region comprising the amino acid sequence of SEQ ID NO: 3, a CDR2 region comprising the amino acid sequence of SEQ ID NO: 4, and a CDR3 region comprising the amino acid sequence of SEQ ID NO: 5; the nanobody may comprise the amino acid sequence of SEQ ID NO: 2 and may be encoded by the nucleotide sequence of SEQ ID NO: 1.
CAR structural components
The CAR further comprises a hinge domain selected from the group consisting of a CD8 hinge, an IgG3s hinge, and an IgG4m hinge, a transmembrane domain (selected from CD8, CD28, and ICOS; e.g., SEQ ID NO: 21), and an intracellular signaling domain comprising 4-1BB and CD3 zeta (e.g., SEQ ID NO: 22); CAR sequences are provided (e.g., SEQ ID NOs: 17, 23, 34, 36) and corresponding nucleotide sequences (e.g., SEQ ID NOs: 33, 35).
Composition comprising a nucleic acid encoding the CAR
A composition comprising a nucleic acid encoding a chimeric antigen receptor (CAR) as recited herein, wherein the CAR comprises a nanobody that specifically binds to the first domain of CDH17 with the CDR sequences of SEQ ID NOs: 3-5.
The independent claims focus on a CAR containing a nanobody that binds the first domain of CDH17 with defined CDR sequences and on compositions encoding such CARs; inventive features emphasize the nanobody specificity and sequences, CAR structural components (hinge, transmembrane, intracellular signaling domains), and nucleic acid compositions encoding the CAR.
Stated Advantages
Isolates antibodies that are shown to bind antigens preferentially expressed in a tumor or cell type-specific manner and that are capable of redirecting CAR T cells to the tumor site in vivo.
Significantly expedites the pace to identify antibodies and their targets to extend CAR T application in various types of cancers.
Provides a nanobody switch/sCAR approach that allows temporal control of CAR T activation, which can reduce on-target-off-tumor toxicity and make CAR T activity reversible and manageable.
Nanobody switches are small with shorter in vivo lifetime and improved tumor microenvironment penetration, and reversible triggering may reduce chronic stimulation and T cell exhaustion.
Documented Applications
Treatment of acute myeloid leukemia (AML) using a CAR T system that targets CD13.
Treatment of neuroendocrine tumors (NETs) and other CDH17-expressing tumors using CAR T systems and ADCs that target CDH17.
Generation of tumor-specific and CAR T cell-compatible nanobodies using the STAR (Sequential Tumor-related Antibody and antigen Retrieving) method.
Use of a switchable CAR system comprising a nanobody fused to a peptide neoepitope (PNE) and a CAR with a PNE-specific scFv to achieve controllable CAR T activation.
Antibody drug conjugates (ADCs) comprising nanobodies linked to a drug, toxin, or radioisotope (e.g., DM1 conjugates) for tumor cell killing and imaging applications.
Use of modified T cells or precursors comprising the disclosed CARs for adoptive cell transfer therapy to treat cancer.
Identification and use of nanobodies and CARs for colorectal cancer as explicitly recited in the disclosure.
Isolation of potent cancer killing or diagnostic antibodies as stated in the background and summary.
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