Modified b cells and methods of use thereof

Abstract

The present invention relates to genetically modified B cells and their uses thereof, for example, for the treatment of a variety of diseases and disorders, including cancer, heart disease, inflammatory disease, muscle wasting disease, neurological disease, and the like. In certain embodiments, the invention relates to an isolated modified B cell (“CAR-B cell”), capable of expressing a chimeric receptor (“CAR-B receptor”), wherein said chimeric receptor comprises (a) an extracellular domain; (b) a transmembrane domain; and (c) a cytoplasmic domain that comprises at least one signaling domain. In various embodiments, the invention comprises an isolated modified B cell, wherein said B cell is capable of expressing and secreting a payload, wherein the payload is not naturally expressed in a B cell or is expressed at higher levels than is naturally expressed in a B cell. In various embodiments, the payload is an antibody or fragment thereof.

1 . A population of modified B cells engineered to express a payload, wherein the B cells comprise a chimeric antigen receptor comprising
 (a) an extracellular antigen binding domain;   (b) a transmembrane domain; and   (c) a cytoplasmic domain comprising
 (i) a cytoplasmic tail comprising a native B cell receptor C-terminus, or 
 (ii) at least one signaling domain, 
   wherein the modified B cells become activated and proliferate upon binding of the antigen binding domain to a target antigen, and   wherein the activation of the modified B cells results in expression and/or increased expression of the payload.   
     
     
         2 . The population of modified B cells of  claim 1 , wherein the payload is not naturally expressed in a B cell or is expressed at a higher level than is naturally expressed in a B cell. 
     
     
         3 . The population of modified B cells of  claim 1 , wherein the payload is a therapeutic protein, a polypeptide, an antibody or fragment thereof, or a nucleic acid. 
     
     
         4 . The population of modified B cells of  claim 1 , wherein the payload is an antibody, antibody fragment, antigen-binding fragment, F(ab′)2, Fab, Fab′, scFv, or Fc fragment. 
     
     
         5 . The population of modified B cells of  claim 1 , wherein the payload is a secreted antibody, antibody fragment, antigen-binding fragment, F(ab′)2, Fab, Fab′, scFv, or Fc fragment. 
     
     
         6 . The population of modified B cells of  claim 1 , wherein the payload is a membrane-bound antibody, antibody fragment, antigen-binding fragment, F(ab′)2, Fab, Fab′, scFv, or Fc fragment. 
     
     
         7 . The population of modified B cells of  claim 1 , wherein the payload is an anti-FAP antibody, an anti-TGF-β antibody, or an anti-BMP antibody. 
     
     
         8 . The population of modified B cells of  claim 1 , wherein the B cells express a payload selected from a group of cytokines, chemokines, T cell costimulatory molecules, and checkpoint molecules, the group consisting of: IL-1, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, IL-18, IL-21, interferon α, interferon β, interferon γ, TSLP, CCL21, FLT3L, XCL1, LIGHT(TNFSF14), OX40L, CD137L, CD40L, ICOSL, anti-CD3 antibody, CD47, TIM4-FC, CXCL13, CCL21, CD80, IFNα A2, LIGHT, 4-1BBL, MDGF (C19orf10), FGF10, PDGF, agrin, TNF-α, GM-CSF, an anti-FAP antibody, a TGF-β trap, decoy or other inhibitory molecule; an anti-BMP antibody; and a BMP trap, decoy or other inhibitory molecule. 
     
     
         9 . The population of modified B cells of  claim 1 , wherein the B cells express more than one payload. 
     
     
         10 . The population of modified B cells of  claim 1 , wherein the antigen binding domain specifically binds a viral protein, bacterial protein or carbohydrate, fungal protein or carbohydrate, protist antigen, or parasitic antigen. 
     
     
         11 . The population of modified B cells of  claim 1 , wherein the target antigen is selected from the group consisting of a hepatitis B antigen, Epstein Barr Virus antigen, HPV antigen, HCV antigen, SARs antigen, and SARS-CoV2 antigen. 
     
     
         12 . The population of modified B cells of  claim 1 , wherein the cytoplasmic domain is selected from the group consisting of: CD79a (Immunoglobulin α), CD79b (Immunoglobulin B), CD40, CD19, CD137, Fcγr2a, MyD88, CD21, Syk, FYN, LYN, P13K, BTK PLCγ2, CD3ζ, and BLNK. 
     
     
         13 . The population of modified B cells of  claim 1 , wherein the extracellular binding domain is a single chain variable fragment (scFv), a full-length antibody or an antibody fragment, or an extracellular domain of a receptor or ligand. 
     
     
         14 . The population of modified B cells of  claim 1 , wherein the at least one signaling domain comprises an immunoreceptor tyrosine-based activation motif (ITAM) of a CD79a. 
     
     
         15 . The population of modified B cells of  claim 1 , wherein the B cells are human B cells. 
     
     
         16 - 27 . (canceled) 
     
     
         28 . A method for treating a subject in need thereof, comprising administering to the subject a population of modified B cells engineered to express a payload, wherein the B cells comprise a chimeric antigen receptor comprising
 (a) an extracellular antigen binding domain;   (b) a transmembrane domain; and   (c) a cytoplasmic domain comprising   (i) a cytoplasmic tail comprising a native B cell receptor C-terminus, or   (ii) at least one signaling domain,   wherein the binding of a target antigen to the antigen binding domain results in expression or increased expression of the payload.   
     
     
         29 . The method of  claim 28 , wherein the payload is not naturally expressed in a B cell or is expressed at a higher level than is naturally expressed in a B cell. 
     
     
         30 . (canceled) 
     
     
         31 . The method of  claim 28 , wherein the payload is an antibody, antibody fragment, antigen-binding fragment, F(ab′)2, Fab, Fab′, scFv, or Fc fragment. 
     
     
         32 - 40 . (canceled) 
     
     
         41 . The method of  claim 28 , wherein the at least one signaling domain comprises an immunoreceptor tyrosine-based activation motif (ITAM) of a CD79a. 
     
     
         42 - 54 . (canceled) 
     
     
         55 . A method of making a population of modified B cells engineered to express a payload comprising:
 obtaining a population of B cells from a patient,   engineering said population of B cells to comprise (contain?) a chimeric antigen receptor said chimeric antigen receptor comprising:
 (a) an extracellular antigen binding domain; 
 (b) a transmembrane domain; and 
 (c) a cytoplasmic domain comprising
 (i) a cytoplasmic tail comprising a native B cell receptor C-terminus, or 
 (ii) at least one signaling domain, 
 
   wherein the modified B cells become activated and proliferate upon binding of the antigen binding domain to a target antigen, and   wherein the activation of the modified B cells results in expression and/or increased expression of the payload.