Compositions and methods for treating cancer with anti-mesothelin immunotherapy
Inventors
ORENTAS, Rimas • Schneider, Dina • Dropulic , Boro • Dimitrov, Dimiter S. • Zhu, Zhongyu
Assignees
Lentigen Technology Inc • US Department of Health and Human Services
Publication Number
US-11970528-B2
Publication Date
2024-04-30
Expiration Date
2038-01-09
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Abstract
Chimeric antigen receptors containing mesothelin antigen binding domains are disclosed. Nucleic acids, recombinant expression vectors, host cells, antigen binding fragments, and pharmaceutical compositions, relating to the chimeric antigen receptors are also disclosed. Methods of treating or preventing cancer in a subject, and methods of making chimeric antigen receptor T cells are also disclosed.
Core Innovation
The invention discloses chimeric antigen receptors (CARs) containing mesothelin antigen binding domains, nucleic acids encoding these receptors, recombinant expression vectors, host cells expressing the CARs, antigen binding fragments, and pharmaceutical compositions. The CARs exhibit high surface expression on transduced T cells, leading to a high degree of cytolysis and in vivo expansion and persistence of the transduced T cells. The methods provided involve treating or preventing cancer in a subject using these CARs and creating CAR T cells.
Cancer, particularly solid tumors, poses a significant challenge due to uncontrolled growth and metastasis, with limited improvement in survival rates over the past decades. Mesothelin is overexpressed in various solid tumors like mesothelioma, ovarian, stomach, lung, pancreatic adenocarcinoma, bile duct carcinoma, and triple negative breast cancer, making it a promising target for immune-based therapies. However, prior attempts to target mesothelin-positive tumors with CARs using mouse-derived antigen binding domains faced challenges such as toxicity, lack of efficacy, and delivery issues. There exists an urgent need for CAR compositions and methods that provide therapeutic efficacy without these shortcomings.
The present invention overcomes these challenges by using entirely human extracellular mesothelin single chain variable fragment (scFv) antigen binding domains in CARs, which improves functional activity and avoids the induction of anti-CAR immune responses seen with mouse-derived sequences. These CARs show superior surface expression, enhanced in vivo persistence, potent cytokine-induced cytolysis, and can be tuned for efficacy and tissue specificity by selecting different binding affinities. The invention also provides detailed descriptions of the CAR components, including extracellular binding domains, transmembrane domains, intracellular signaling domains, and linker or spacer domains, as well as methods of use, making it a comprehensive advancement in anti-mesothelin immunotherapy.
Claims Coverage
The patent claims cover methods of using chimeric antigen receptor (CAR) T cells or natural killer (NK) cells comprising CARs with specific mesothelin antigen binding domains for anti-tumor immunity and cancer treatment, detailing the molecular features and functional domains of the CAR constructs.
Method of providing anti-tumor immunity using CAR-expressing T cells or NK cells
Administering to a human subject having a tumor an effective amount of isolated T cells or NK cells that comprise vectors encoding a CAR with at least one extracellular mesothelin antigen binding domain having the amino acid sequence of SEQ ID NO. 4, 6, or 8, including at least one linker or spacer domain, one transmembrane domain, and one intracellular signaling domain, thereby providing anti-tumor immunity.
Use of specified transmembrane domains in CARs
The CARs comprise transmembrane domains selected from proteins including alpha, beta, or zeta chain of T-cell receptor, CD8, CD28, CD3 epsilon, CD45, CD4, CD5, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, and CD154.
Connection of domains by linker or spacer
The mesothelin antigen binding domain and the intracellular signaling domain in the CAR are connected to the transmembrane domain via at least one linker or spacer domain, which can be derived from extracellular domains of CD8 or CD28.
Nucleic acid encoding for specific mesothelin binding sequences
The nucleic acid sequence encoding the mesothelin antigen binding domain includes SEQ ID NO: 3, 5, or 7, or sequences with 85% to 99% identity thereto.
Intracellular signaling domain composition
The intracellular signaling domain includes a signaling domain of CD3 zeta and may also comprise a costimulatory domain, a primary signaling domain, or both.
Costimulatory domain characteristics
The costimulatory domain comprises functional signaling domains of proteins selected from OX40, CD70, CD27, CD28, CD5, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), DAP10, DAP12, and 4-1BB (CD137).
Use of specific amino acid sequences for CARs
The CARs comprise the amino acid sequences of SEQ ID NO: 14, SEQ ID NO: 16, or SEQ ID NO: 18.
Method of treating cancer using CAR-expressing T or NK cells
Administering to a human subject an effective amount of isolated T cells or NK cells expressing CARs as described above, comprising mesothelin antigen binding domains of SEQ ID NO: 4, 6, or 8, for treatment of cancer.
The claims cover methods of using CARs with fully human mesothelin antigen binding domains in T cells or NK cells for anti-tumor immunity and cancer treatment, focusing on specific compositions of CAR domains including antigen binding, transmembrane, linker, and intracellular signaling regions, as well as specified nucleic acid and amino acid sequences.
Stated Advantages
The CARs exhibit high surface expression on transduced T cells leading to high in vivo proliferation and persistence.
Use of fully human mesothelin antigen binding domains avoids anti-CAR immune responses and eliminates rapid CAR T cell elimination associated with mouse-derived sequences.
CARs demonstrate a high degree of cytokine-induced cytolysis and potent cytotoxic activity against mesothelin-positive tumor cells.
Ability to tune CAR affinity to balance efficacy and tissue specificity, potentially reducing on-target off-tumor toxicity and bystander cell killing.
Documented Applications
Treatment or prevention of cancer in a subject by administration of CAR-expressing T cells or NK cells targeting mesothelin.
Use in treating various hematological cancers including acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), lymphoma, multiple myeloma.
Use in treating solid tumors such as mesothelioma, ovarian, pancreatic, stomach, lung, bile duct carcinoma, and triple negative breast cancer.
Methods for generating CAR T cells or RNA-engineered cells expressing anti-mesothelin CARs for adoptive cell therapy.
Diagnostic methods for detecting diseases associated with mesothelin expression using anti-mesothelin antibodies or fragments.
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