Mhc class ii t-cell modulatory multimeric polypeptides for treating type 1 diabetes mellitus (t1d) and methods of use thereof

Abstract

The present disclosure provides T-cell modulatory multimeric polypeptides (TMMPs) comprising a type 1 diabetes (T1D)-associated peptide epitope, MHC class II polypeptides, and one or more immunomodulatory polypeptides. A TMMP of the present disclosure is useful for modulating activity of a T cell. Thus, the present disclosure provides compositions and methods for modulating the activity of T cells, as well as compositions and methods for treating persons who have T1D.

1 .- 66 . (canceled) 
     
     
         67 . A T-cell modulatory multimeric polypeptide (TMMP) comprising at least one heterodimer, wherein each heterodimer comprises:
 a) a first polypeptide comprising:
 i) a Type 1 Diabetes associated peptide (T1D peptide) having a length of from 10-20 amino acids; and 
 ii) a first major histocompatibility complex (MHC) class II polypeptide, wherein the first MHC class II polypeptide is an MHC class II β polypeptide; and 
 iii) a cysteine (Cys)-containing linker that links the T1D peptide to the MHC class II β polypeptide; and 
   b) a second polypeptide comprising:
 i) one or more immunomodulatory polypeptides, wherein at least one of the one or more immunomodulatory polypeptides is a PD-L1 polypeptide or a FasL polypeptide; 
 ii) a second MHC class II polypeptide, wherein the second MHC class II polypeptide is an MHC class II α polypeptide that comprises a Cys at position 72 or 75 based on the amino acid numbering depicted in  FIG.  13 A ; and 
 iii) an immunoglobulin (Ig) Fc polypeptide, 
   wherein the first and the second polypeptide of the heterodimer are covalently linked to one another via a disulfide bond between the Cys in the Cys-containing linker and the Cys at position 72 or 75 of the MHC class II α polypeptide, and   wherein the TMMP presents a Type 1 Diabetes-associated epitope capable of being bound by a T-cell receptor on a CD4+ T cell.   
     
     
         68 . A TMMP of  claim 67 , wherein
 a1) the first polypeptide comprises, in order from N-terminus to C-terminus:
 i) the T1D peptide; 
 ii) the Cys-containing linker; and 
 ii) the MHC class II β polypeptide; and 
   b1) the second polypeptide comprises, in order from N-terminus to C-terminus:
 i) the one or more immunomodulatory polypeptides; 
 ii) the MHC class II α polypeptide; and 
 iii) the Ig Fc polypeptide, 
   wherein the components of the second polypeptide optionally may be joined by one or more linkers.   
     
     
         69 . A TMMP of  claim 68 , wherein the MHC class II α polypeptide comprises an amino acid sequence having at least 95% amino acid sequence identity to a DRA1*01:01 polypeptide; and the MHC class II R polypeptide comprises an amino acid sequence having at least 95% amino acid sequence identity to a DRB1*04:01 polypeptide. 
     
     
         70 . A TMMP of  claim 69 , wherein the TMMP comprises a PD-L1 polypeptide, and wherein the PD-L1 immunomodulatory polypeptide comprises an amino acid sequence having at least 95% amino acid sequence identity to the PD-L1 amino acid sequence of SEQ ID NO:99, and has a length of about 220 amino acids. 
     
     
         71 . A TMMP of  claim 70 , wherein the T1D peptide comprises is a proinsulin peptide or a glutamic acid decarboxylase (GAD) peptide. 
     
     
         72 . A TMMP of  claim 71 , wherein the T1D peptide:
 a) is a proinsulin peptide selected from SLQPLALEGSLQKRG (SEQ ID NO:94; proIns 76-90), SLQPLALEGSLQSRG (SEQ ID NO:90; proIns 76-90; K88S), GAGSLQPLALEGSLQKRG (SEQ ID NO:93; proIns 73-90) and GSLQPLALEGSLQSRGIV (SEQ ID NO:91; proIns 75-92(K88S); or   b) is a GAD peptide selected from NFFRMVISNPAAT (SEQ ID NO:87; GAD65 555-567) and NFIRMVISNPAAT (SEQ ID NO:88; GAD65 555-567; F557I).   
     
     
         73 . A TMMP of  claim 71 , wherein the MHC class II α polypeptide comprises a K75C substitution. 
     
     
         74 . A TMMP comprising two heterodimers according to  claim 73 , wherein the heterodimers are disulfide bonded to each other though their respective Ig Fc components. 
     
     
         75 . A pharmaceutical composition comprising a TMMP of  claim 74 . 
     
     
         76 . One or more nucleic acids comprising nucleotide sequences encoding a TMMP of  claim 73 . 
     
     
         77 . A method of making a TMMP comprising the step of culturing a host cell according to  claim 76  under conditions in which the host cell produces the TMMP. 
     
     
         78 . A method of selectively modulating the activity of CD4 +  T cells specific for a type 1 diabetes-associated epitope in an individual, the method comprising administering the TMMP of  claim 74  to the individual an effective amount of the TMMP. 
     
     
         79 . A method of reducing the number and/or activity of CD4 +  and/or CD8 +  self-reactive T cells specific for a type 1 diabetes-associated epitope in an individual, the method comprising administering an effective amount of the TMMP of  claim 74  to the individual. 
     
     
         80 . A method of treating type 1 diabetes (T1D) in an individual, the method comprising administering to an individual in need thereof an effective amount of the TMMP of  claim 74 . 
     
     
         81 . An antigen-presenting polypeptide (APP) comprising:
 a) a first polypeptide comprising:
 i) a peptide that displays a Type 1 Diabetes-associated epitope capable of being bound by a T-cell receptor (a “T1D peptide”); and 
 ii) a first major histocompatibility complex (MHC) class II polypeptide; and 
 iii) optionally a linker that links the T1D peptide to the first MHC class II polypeptide; and 
   b) a second polypeptide comprising a second MHC class II polypeptide,   wherein the first and the second polypeptide of the heterodimer are covalently linked to one another via at least one disulfide bond,   wherein one of the polypeptides of the heterodimer comprises an immunoglobulin (Ig) Fc polypeptide, optionally joined to the first or second polypeptide via a linker, and   wherein the APP does not include an immunomodulatory polypeptide.   
     
     
         82 . One or more nucleic acids comprising nucleotide sequences encoding an APP of  claim 81 . 
     
     
         83 . A method of making an APP comprising culturing a host cell comprising the one or more nucleic acids of  claim 82  under conditions in which the host cell produces the TMMP. 
     
     
         84 . A method of reducing the number and/or activity of CD4 +  and/or CD8 +  self-reactive T cells specific for a type 1 diabetes-associated epitope in an individual, the method comprising administering an effective amount of the APP of  claim 81  to the individual. 
     
     
         85 . A method of treating type 1 diabetes (T1D) in an individual, the method comprising administering to an individual in need thereof an effective amount of the APP of  claim 81 .