ROR-1 specific chimeric antigen receptors and uses thereof
Inventors
SHAH, Rutul R. • Chen, ChangHung • BOLINGER, Cheryl G. • KURELLA, Vinodhbabu • Wesa, Amy
Assignees
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Publication Number
US-12275772-B2
Publication Date
2025-04-15
Expiration Date
Abstract
Provided herein are chimeric antigen receptors (CARs) for cancer therapy, and more particularly, CARs containing a scFv from an anti-ROR-1 monoclonal antibody. Provided are immune effector cells containing such CARs, and methods of treating proliferative disorders.
Core Innovation
The disclosure describes a chimeric antigen receptor (CAR) that targets ROR-1 and includes a ROR-1 antigen binding domain comprising a first polypeptide and a second polypeptide with specified amino acid sequences, together with a transmembrane domain and a CD3 zeta signaling domain. The CAR is presented as suitable for expression on immune effector cells for cancer therapy.
The CAR architecture is described in multiple configurations by selecting different spacer/stalk domains, transmembrane domains, and cytoplasmic signaling domains. The cytoplasmic signaling may include CD3 zeta alone, or CD3 zeta with costimulatory signaling domains such as 4-1BB and/or CD28, and the disclosure also describes sequence-identity ranges of CAR subdomains relative to multiple SEQ ID NOs.
The disclosure further describes engineered immune effector cells and nucleic acid delivery systems for expressing the ROR-1 CAR, including viral vectors, lentivirus/retrovirus, Sleeping Beauty transposon systems, non-viral DNA transposons, and integrating/viral vectors such as PiggyBac. Additional elements include optional cytokines such as membrane-bound IL-15, cell tag and kill switch concepts, and gene-switch and ligand-inducible control systems including an ecdysone receptor-based system and related transcriptional components.
The described configurations and applications are directed to treating cancers and proliferative disorders associated with ROR-1 expression, including CAR-T stimulation and expansion and target tumor cell killing.
Claims Coverage
The claim coverage centers on one independent claim for a nucleic acid encoding a ROR-1 CAR and multiple dependent claim refinements. In total, the claim set describes seven inventive features, including defined antigen-binding polypeptides, transmembrane and CD3 zeta signaling domains, sequence-identity constraints, spacer and costimulatory features, and immune effector cell and therapeutic use features.
ROR-1 antigen binding domain with defined first and second polypeptides
A nucleic acid encoding a CAR comprising a ROR-1 antigen binding domain including a first polypeptide comprising the amino acid sequence of SEQ ID NO: 36 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 17.
Transmembrane domain and CD3 zeta signaling domain
A nucleic acid encoding a CAR comprising a transmembrane domain and a CD3 zeta signaling domain.
Sequence identity constraint for CD3 zeta signaling domain
The nucleic acid in which the CD3 zeta signaling domain encodes a polypeptide having at least 90% identity to the amino acid sequence of SEQ ID NO: 93.
CAR further comprising a spacer
The nucleic acid encoding the CAR further comprising a spacer.
Multiple CAR subdomain amino-acid identity constraints including spacer, transmembrane, CD28 costimulation, and CD3 zeta
The nucleic acid encoding the CAR includes specified spacer, transmembrane, CD28 costimulatory signaling, and CD3 zeta signaling polypeptide domains, each having at least 90% amino-acid sequence identity to corresponding SEQ ID NOs (85, 87, 90, and 93).
Immune effector cell further comprising a cell tag and a cytokine
An immune effector cell further comprising a cell tag and a cytokine.
Treating cancer with engineered T-cells expressing a CAR and membrane-bound IL-15
A method for treating cancer in a human administering one or more doses of engineered T-cells expressing a CAR encoded by the nucleic acid of claim 1 and a membrane-bound IL-15 to a subject whose cancer expresses ROR-1.
Across the claim set, coverage centers on a nucleic-acid-encoded ROR-1 CAR with defined antigen-binding polypeptides (SEQ ID NO: 36 and SEQ ID NO: 17) plus a transmembrane domain and a CD3 zeta signaling domain. Dependent claims further include sequence-identity constraints, a spacer, CD28 costimulatory signaling, cell tags and cytokines, and therapeutic treatment of cancer expressing ROR-1 with engineered T-cells and membrane-bound IL-15.
Stated Advantages
Tumor growth inhibition is reported as dose-dependent.
CAR-T persistence and expansion are measured by flow cytometry.
Multi-parameter readouts include CD107, IFN-γ, and TNF-α.
Documented Applications
In vitro cytotoxicity testing of CAR-T/mbIL-15 configurations against SKOV3_fLUC and JeKo-1_fLUC tumor lines.
In vivo NSG xenograft study with JeKo-1 tumor cells.
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