Conditionally active heterodimeric polypeptides and methods of use thereof

Abstract

The present disclosure provides conditionally active, heterodimeric polypeptides. The conditionally active, heterodimeric polypeptides are active in the presence of a dimerizing agent that induces dimerization of the polypeptides of the heterodimer. A conditionally active, heterodimeric polypeptide of the present disclosure is useful in a variety of research and treatment methods, which are also provided.

1 - 91 . (canceled) 
     
     
         92 . An immune cell comprising one or more nucleic acids comprising:
 (a) a first nucleotide sequence encoding a first polypeptide of a chimeric antigen receptor (CAR) heterodimer comprising an antigen binding domain and a first member of a dimerization pair; and   (b) a second nucleotide sequence encoding a second polypeptide of the CAR heterodimer comprising an intracellular signaling domain and a second member of the dimerization pair, wherein the first polypeptide of the CAR heterodimer, the second polypeptide of the CAR heterodimer or both comprise a co-stimulatory domain;   wherein one member of the dimerization pair comprises a ligand-binding domain (LBD) of a nuclear hormone receptor but not the DNA binding domain of the nuclear hormone receptor and the other member of the dimerization pair comprises at least two copies of a dimerization sequence of a co-regulator of the nuclear hormone receptor, wherein the dimerization sequence is in the range of 16 amino acids to 50 amino acids in length; and   wherein the first and polypeptides dimerize in the presence of the nuclear hormone.   
     
     
         93 . The cell according to  claim 92 , wherein the first polypeptide or the second polypeptide of the CAR heterodimer comprises three copies of the co-regulator of the nuclear hormone receptor. 
     
     
         94 . The cell according to  claim 92 , wherein the first polypeptide and the second polypeptide of the CAR heterodimer each comprise the same co-stimulatory domain. 
     
     
         95 . The cell according to  claim 92 , wherein the first polypeptide and the second polypeptide of the CAR heterodimer comprise different co-stimulatory domains. 
     
     
         96 . The cell according to  claim 92 , wherein the first polypeptide of the CAR heterodimer, the second polypeptide of the CAR heterodimer or both comprise two or more co-stimulatory domains. 
     
     
         97 . The cell according to  claim 92 , wherein the co-stimulatory domain is selected from the group consisting of: a 4-1BB (CD137) co-stimulatory domain, a CD28 co-stimulatory domain, a ICOS co-stimulatory domain, an OX-40 co-stimulatory domain, a BTLA co-stimulatory domain, a CD27 co-stimulatory domain, a CD30 co-stimulatory domain, a GITR co-stimulatory domain, and a HVEM co-stimulatory domain. 
     
     
         98 . The cell according to  claim 92 , wherein the first polypeptide of the CAR heterodimer comprises a transmembrane domain. 
     
     
         99 . The cell according to  claim 92 , wherein the second polypeptide of the CAR heterodimer comprises a transmembrane domain. 
     
     
         100 . The cell according to  claim 92 , wherein the antigen binding domain is a scFv or nanobody. 
     
     
         101 . The cell according to  claim 100 , wherein the scFv or nanobody specifically binds a cancer associated antigen. 
     
     
         102 . The cell according to  claim 92 , wherein the intracellular signaling domain comprises at least one immunoreceptor tyrosine-based activation motif (ITAM). 
     
     
         103 . The cell according to  claim 102 , wherein the intracellular signaling domain comprises an amino acid sequence having at least 85% sequence identity to one or more of SEQ ID NOs: 566-588. 
     
     
         104 . The cell according to  claim 92 , wherein the intracellular signaling domain comprises an amino acid sequence having at least 85% sequence identity to one or more of SEQ ID NOs: 589-593. 
     
     
         105 . The cell according to  claim 92 , wherein the cell is a T lymphocyte or progenitor thereof. 
     
     
         106 . A method of modulating an activity of an immune cell, comprising contacting the immune cell of  claim 92  with the nuclear hormone an analog thereof, wherein in the presence of the nuclear hormone an analog thereof the first and second polypeptides dimerize. 
     
     
         107 . A method of treating cancer in an individual, comprising administering to the individual:
 a) an immune cell according to  claim 92 ; and   b) an effective amount of the nuclear hormone an analog thereof;   wherein the immune cell is activated in the individual by binding to a target antigen on a cancer cell.