Regulatable chimeric antigen receptor

A regulatable chimeric antigen receptor (RCAR), wherein the RCAR comprises:

• a) an inhibitory counter ligand binding member comprising:
• an inhibitory counter ligand binding domain from a molecule chosen from B7-H1, B7-1, CD160, P1H, 2B4, PD1, TIM3, CEACAM, LAGS, TIGIT, CTLA-4, BTLA, LAIR1, or TGF-beta receptor,
• a transmembrane domain or a membrane anchoring domain, and
• a first switch domain of a FKBP-FRB based switch;
• and
• b) an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a second switch domain of a FKBP-FRB based switch, wherein:
• the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations.

A regulatable chimeric antigen receptor (RCAR), wherein the RCAR comprises:

• a) a costimulatory extracellular domain (ECD) member comprising:
• a costimulatory ECD domain from a molecule chosen from ICOS, CD28, VEM, LIGHT, CD40L, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, or CD226,
• a transmembrane domain or a membrane anchoring domain, and
• a first switch domain of a FKBP-FRB based switch;
• b) an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a second switch domain of a FKBP-FRB based switch, wherein:
• the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations.

A nucleic acid encoding a regulatable chimeric antigen receptor (RCAR), wherein the RCAR comprises:

• a) an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a first switch domain of a FKBP-FRB based switch;
• and
• b) an antigen binding member comprising:
• an antigen binding domain,
• a transmembrane domain or a membrane anchoring domain, and
• a second switch domain of a FKBP-FRB based switch, wherein:
• the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations chosen from an E2032 mutation, a T2098 mutation, or an E2032 and a T2098 mutation.

A nucleic acid encoding an RCAR comprising:

• (a) a sequence encoding an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a first switch domain of a FKBP-FRB based switch;
• (b) a sequence encoding an antigen binding member comprising:
• an antigen binding domain,
• a transmembrane domain or a membrane anchoring domain, and
• a second switch domain of a FKBP-FRB based switch;
• and
• (c) a sequence encoding a cleavable peptide or an IRES,
• wherein:
• i) (a), (b), and (c) are present on a single nucleic acid molecule, are transcribed as a single transcription product, and are configured as follows:
• a promoter is operably linked to (a), (b), and (c), wherein (c) encodes a cleavable peptide, and (c) is disposed between (a) and (b);
• or
• ii) (a), (b), and (c) are present on a single nucleic acid molecule, are transcribed as a single transcription product, and are configured as follows:
• a promoter is operably linked to (a), (b), and (c), wherein (c) encodes an IRES, and (c) is disposed between (a) and (b), and wherein:
• the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations selected from an E2032 mutation, a T2098 mutation, or an E2032 and a T2098 mutation.

A method of treating a subject comprising:

• (a) administering to the subject an effective amount of an RCARX cell, wherein the RCARX cell comprises:
• an RCAR, or a nucleic acid encoding the RCAR;
• and
• (b) administering to the subject a dimerization molecule,
• wherein the RCAR comprises:
• A) an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a first switch domain;
• and
• B) an antigen binding member comprising:
• an antigen binding domain,
• a transmembrane domain or a membrane anchoring domain, and
• a second switch domain,
• wherein the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations chosen from an E2032 mutation, a T2098 mutation, or an E2032 and a T2098 mutation, wherein:
• the subject has a cancer, a viral disease, or an autoimmune disease.

A nucleic acid encoding a regulatable chimeric antigen receptor (RCAR), wherein the RCAR comprises:

• a) an inhibitory counter ligand binding member comprising:
• an inhibitory counter ligand binding domain from a molecule chosen from B7-H1, B7-1, CD160, P1H, 2B4, PD1, TIM3, CEACAM, LAG3, TIGIT, CTLA-4, BTLA, LAIR1, or TGF-beta receptor,
• a transmembrane domain or a membrane anchoring domain, and
• a first switch domain of a FKBP-FRB based switch;
• and
• b) an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a second switch domain of a FKBP-FRB based switch, wherein:
• the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations.

A nucleic acid encoding a regulatable chimeric antigen receptor (RCAR), wherein the RCAR comprises:

• a) a costimulatory extracellular domain (ECD) member comprising:
• a costimulatory ECD domain from a molecule chosen from ICOS, CD28, VEM, LIGHT, CD40L, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, or CD226,
• a transmembrane domain or a membrane anchoring domain, and
• a first switch domain of a FKBP-FRB based switch;
• b) an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a second switch domain of a FKBP-FRB based switch, wherein:
• the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations.

A regulatable chimeric antigen receptor (RCAR), wherein the RCAR comprises:

• a) a costimulatory extracellular domain (ECD) member comprising:
• a costimulatory ECD domain from a molecule chosen from ICOS, CD28, CD27, HVEM, LIGHT, CD40L, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, or CD226,
• a transmembrane domain or a membrane anchoring domain, and
• a first switch domain of a FKBP-FRB based switch;
• b) an intracellular signaling member comprising:
• an intracellular signaling domain, and
• a second switch domain of a FKBP-FRB based switch, wherein:
• the first switch domain comprises a FRB binding fragment or analog of FKBP and the second switch domain comprises a FKBP binding fragment or analog of FRB, or the first switch domain comprises a FKBP binding fragment or analog of FRB and the second switch domain comprises a FRB binding fragment or analog of FKBP, wherein:
• the FKBP binding fragment or analog of FRB comprises one or more mutations chosen from an E2032 mutation, a T2098 mutation, or an E2032 and a T2098 mutation.