The present application provides single-domain antibodies targeting BCMA, and chimeric antigen receptors (such as monovalent CAR, and multivalent CAR including bi-epitope CAR) comprising one or more anti-BCMA single-domain antibodies. Further provided are engineered immune effector cells (such as T cells) comprising the chimeric antigen receptors. Pharmaceutical compositions, kits and methods of treating cancer are also provided.

The current application fulfills a need for methods of making bi-specific CAR T-cells that have high efficacy. Aspects of the disclosure relate to a method for manufacturing a CD19/CD20 bi-specific chimeric antigen receptor (CAR) T cell comprising the ordered steps of: (a) providing a composition comprising a population of cells comprising T cells; (b) contacting the composition comprising the population of cells comprising T cells with one or more of a transactivating composition, IL-2, and/or IL-15; (c) transducing the cell composition from (b) with a CD19/CD20 bi-specific CAR nucleic acid; and (d) removing the transactivating composition from the cell composition of (c). Further aspects relate to a population of CD19/CD20 bi-specific CAR-T cells produced by the methods of the disclosure. Yet further aspects describe a method for treating a subject for B-cell lymphoma comprising administering cells of the disclosure.

The present invention provides compositions and methods for transducing cells (e.g. T cells or immune cells). Also provided herein are methods of treating a disease in a subject in need thereof.

The invention is directed to a bispecific chimeric antigen receptor, comprising: (a) at least two antigen-specific targeting regions; (b) an extracellular spacer domain; (c) a transmembrane domain; (d) at least one co-stimulatory domain; and (e) an intracellular signaling domain, wherein each antigen-specific targeting region comprises an antigen-specific single chain Fv (scFv) fragment, and binds a different antigen, and wherein the bispecific chimeric antigen receptor is co-expressed with a therapeutic control. The invention also provides methods and uses of the bispecific chimeric antigen receptors.

Aspects of the disclosure include methods of effectively treating a relapsed/refractory B cell malignancy in a patient in need thereof who has previously been treated with and ultimately failed at least one, two, or at least three previous therapies. In one example. a method comprises administering to the subject one or more doses comprising a therapeutically effective amount of anti-CD20 CAR T cells that express a chimeric antigen receptor (CAR) comprising a binding domain that specifically binds to CD20 on a malignant B cell to the subject. Following administration, the subject methods optionally further include monitoring for one or more biomarkers of cell activation and/or therapeutic efficacy that inform the need for and/or type of follow-on treatment regimen.

An aptamer-based switch technology is provided that enhances control of the use of chimeric antigen receptor (CAR)-related immunotherapies. The aptamer-based switch utilizes a synthetic bridge molecule containing a target-binding aptamer bound through a linker to a CAR-binding aptamer. A system containing a CAR and a corresponding aptameric bridge provides an immunotherapy platform that: (i) can be targeted to any desired antigen by choosing the target-binding aptamer of the bridge. (ii) can be redirected from one target to another by changing the target-binding aptamer: (iii) can be dosed according to the changing needs of an individual patient overtime by altering the administration protocol for the bridge: (iv) can be switched on or off quickly or gradually: (v) can be used as a companion diagnostic for a specific CAR therapy: (vi) can be integrated with either in vivo or ex vivo CAR expression: (vii) is non-immunogenic; and (viii) has low production costs.

Disclosed herein are chimeric antigen receptor effector cells (CAR-ECs) and CAR-EC switches. The switchable CAR-ECs are generally T cells. The one or more chimeric antigen receptors may recognize a peptidic antigen on the CAR-EC switch. The CAR-ECs and switches may be used for the treatment of a condition in a subject in need thereof.

A CD20-OR-CD19 chimeric antigen receptor (CAR) protein construct is provided. Also provided are nucleic acids encoding the CD20-OR-CD19 CAR; and methods of use, e.g. in the treatment of B cell malignancies. The CD20-OR-CD19 CAR of the invention is a bispecific CAR that can trigger T-cell activation upon detection of either CD19 or CD20 (or both). It is a single molecule that confers two-input recognition capability upon human T cells engineered to stably express this CAR.

The present disclosure provides methods for controllable stimulation of genetically engineered lymphocytes. Compositions and methods of treatment are also provided.

A CD20-OR-CD19 chimeric antigen receptor (CAR) protein construct is provided. Also provided are nucleic acids encoding the CD20-OR-CD19 CAR; and methods of use, e.g. in the treatment of B cell malignancies. The CD20-OR-CD19 CAR of the invention is a bispecific CAR that can trigger T-cell activation upon detection of either CD19 or CD20 (or both). It is a single molecule that confers two-input recognition capability upon human T cells engineered to stably express this CAR.