Novel cellular receptors and uses thereof

Abstract

Described herein are polypeptides, systems, and methods that relate to using domains that bind specifically to a biotinylamide to control receptor and cellular activity.

What is claimed herein is: 
     
         1 . A synthetic signaling system comprising a cell, the cell comprising a small molecule-controlled signaling polypeptide comprising:
 a domain that binds specifically to a small molecule;   a transmembrane domain; and   an intracellular CAR stimulatory domain.   
     
     
         2 . The system of  claim 1 , wherein the domain that binds specifically to a small molecule and the intracellular CAR stimulatory domain are not found as part of the same polypeptide in nature. 
     
     
         3 . The system of  claim 1 , wherein the CAR stimulatory domain is a CD3ζ signaling domain. 
     
     
         4 . The system of  claim 3 , wherein the small molecule-controlled signaling polypeptide further comprises a CAR co-stimulatory domain. 
     
     
         5 . The system of  claim 4 , wherein the CAR co-stimulatory domain is a CD28 signaling domain. 
     
     
         6 . The system of  claim 1 , wherein the small molecule-controlled signaling polypeptide further comprises a CAR co-stimulatory domain. 
     
     
         7 . The system of  claim 6 , wherein the CAR co-stimulatory domain is a CD28 signaling domain. 
     
     
         8 . The system of  claim 1 , wherein the domain that binds specifically to a small molecule is extracellular. 
     
     
         9 . The system of  claim 1 , wherein the small molecule is biotin, biotinidase-resistant biotin, a biotinylamide, bitoinidase-resistant biotinylamide, fluorescein, digoxigenin, or fluorescein isothiocyanate (FITC). 
     
     
         10 . The system of  claim 1 , wherein the domain that binds specifically to a small molecule is an antibody or antibody reagent. 
     
     
         11 . The system of  claim 10 , wherein the antibody reagent is a scFv. 
     
     
         12 . The system of  claim 10 , wherein the antibody reagent is a scFab. 
     
     
         13 . The system of  claim 10 , wherein the small molecule is biotin, biotinidase-resistant biotin, a biotinylamide, or a bitoinidase-resistant biotinylamide, and the antibody reagent comprises the 6 CDRs of SEQ ID NOs: 4-9. 
     
     
         14 . The system of  claim 10 , wherein the small molecule is biotin, biotinidase-resistant biotin, a biotinylamide, or a bitoinidase-resistant biotinylamide, and the antibody reagent comprises SEQ ID NOs: 1 and 2. 
     
     
         15 . The system of  claim 10 , wherein the small molecule is biotin, biotinidase-resistant biotin, a biotinylamide, or a bitoinidase-resistant biotinylamide, and the antibody reagent comprises amino acids 1-119 of SEQ ID NO: 1 and amino acids 1-117 of SEQ ID NO: 2. 
     
     
         16 . The system of  claim 1 , further comprising a polypeptide comprising at least one of:
 a small molecule acceptor peptide and a small molecule.   
     
     
         17 . The system of  claim 16 , wherein the polypeptide comprising at least one of:
 a small molecule acceptor peptide and a small molecule   
       is a soluble molecule further comprising an antibody or antibody reagent. 
     
     
         18 . The system of  claim 16 , further comprising a further soluble molecule comprising a small molecule. 
     
     
         19 . The system of  claim 16 , wherein the polypeptide comprising at least one of:
 a small molecule acceptor peptide and a small molecule   
       is a surface-attached molecule. 
     
     
         20 . The system of  claim 19 , wherein the surface-attached polypeptide comprising at least one of:
 a small molecule acceptor peptide and a small molecule further comprises a binding domain specific for a target.   
     
     
         21 . The system of  claim 19 , further comprising a soluble molecule comprising a small molecule. 
     
     
         22 . A method of treating a subject in need of immunotherapy, the method comprising administering to the subject the system of  claim 1  or a nucleic acid encoding a small molecule-controlled signaling polypeptide comprising:
 a domain that binds specifically to a small molecule; 
 a transmembrane domain; and 
 an intracellular CAR stimulatory domain. 
 
     
     
         23 . The method of  claim 22 , wherein the immune cell is a T cell. 
     
     
         24 . The method of  claim 22 , further comprising administering to the subject a surface-attached polypeptide comprising at least one of a small molecule acceptor peptide and a small molecule. 
     
     
         25 . The method of  claim 22 , further comprising administering to the subject a polypeptide comprising i) at least one of a small molecule acceptor peptide and a small molecule and ii) at least one of a binding domain specific for a target, an antibody, and an antibody reagent. 
     
     
         26 . The method of  claim 25 , wherein the target of the binding domain is a marker on the surface of a diseased cell and/or the antibody or antibody reagent binds specifically to a marker on the surface of a diseased cell. 
     
     
         27 . The method of  claim 26 , wherein the diseased cell is a cancer cell. 
     
     
         28 . The method of  claim 26 , wherein the target is a cell surface marker specific to cancer cells or found on the surface of a cancer cell. 
     
     
         29 . The method of  claim 24 , further comprising administering to the subject a soluble molecule comprising a small molecule or a soluble molecule comprising a small molecule. 
     
     
         30 . A synthetic signaling system comprising:
 a cell, the cell comprising a small molecule-controlled signaling polypeptide comprising:
 a domain that binds specifically to a small molecule; 
 a transmembrane domain; and 
 an intracellular Notch receptor signaling domain; and 
   an extracellular soluble molecule comprising:
 a small molecule; and 
 an antibody or antibody reagent.