Methods and compositions for increasing the suppressive function of regulatory t-cells (tregs)

Abstract

Methods and compositions for increasing the suppressive function of regulatory T-cells (Tregs) are provided.

What is claimed is: 
     
         1 . A method of reducing or eliminating the vimentin protein and/or the RLTPR protein and/or the PKC-θ protein in Treg cells, comprising:
 contacting the Treg cells with a vimentin-specific and/or a RLTPR-specific and/or a PKC-θ-specific inhibitory nucleic acid molecule. 
 
     
     
         2 . The method of  claim 1 , wherein the vimentin-specific inhibitory nucleic acid molecule is complementary to at least a portion of the sequence shown in SEQ ID NO: 1. 
     
     
         3 . The method of  claim 1 , wherein the RLTPR-specific inhibitor nucleic acid is complementary to at least a portion of the sequence shown in SEQ ID NO: 5. 
     
     
         4 . The method of  claim 1 , wherein the PKC-θ-specific inhibitory nucleic acid molecule is complementary to at least a portion of the sequence shown in SEQ ID NO: 9. 
     
     
         5 . The method of  claim 1 , wherein the vimentin-specific and/or the RLTPR-specific and/or the PKC-θ-specific inhibitory nucleic acid molecule is a RNAi nucleic acid molecule. 
     
     
         6 . The method of  claim 1 , wherein the vimentin-specific and/or the RLTPR-specific and/or the PKC-θ-specific inhibitory nucleic acid molecule is an antisense nucleic acid molecule. 
     
     
         7 . The method of  claim 1 , wherein the vimentin-specific and/or the RLTPR-specific and/or the PKC-θ-specific inhibitor nucleic acid molecule is a siRNA nucleic acid molecule. 
     
     
         8 . The method of  claim 1 , wherein the vimentin-specific inhibitory nucleic acid molecule has the sequence shown in SEQ ID NO: 13, 14, 15 or 16. 
     
     
         9 . The method of  claim 1 , wherein the Treg cells are contacted in vitro. 
     
     
         10 . The method of  claim 1 , wherein the Treg cells are contacted in situ. 
     
     
         11 . The method of  claim 1 , wherein the Treg cells are contacted in vivo in an individual who has received or is receiving a bone marrow transplant. 
     
     
         12 . The method of  claim 1 , wherein the Treg cells exhibit a phenotype of at least one of the following:
 reduced PKC-θ auto-phosphorylation at Ser676;   improved ability to suppress CD4+ and CD8+ Tcon proliferation;   increased surface expression of Nrp1;   increased surface expression of Lag3;   increased basal and maximal oxygen consumption rate (OCR);   increased BoDipy C1-C12  uptake;   increased expression of CD71;   increased expression of CD98;   increased expression of CPT1a; or   reduced activity of mTORC2,   relative to Tregs that lack the vimentin-specific and/or the RLTPR-specific and/or the PKC-θ-specific inhibitory nucleic acid molecule.   
     
     
         13 . A method of increasing or augmenting the suppressor cell potency of Treg cells, comprising:
 reducing or eliminating vimentin and/or RLTPR and/or PKC-θ in the Treg cells.   
     
     
         14 . The method of  claim 13 , wherein reducing or eliminating the vimentin and/or the RLTPR and/or the PKC-θ in the Treg cells comprising contacting the Treg cells with a moiety selected from the group consisting of a nucleic acid, a nuclease, an antibody, a ligand, a peptide, a drug, a chemical, or a small molecule. 
     
     
         15 . The method of  claim 14 , wherein the nucleic acid is a vimentin-specific and/or a RLTPR-specific and/or a PKC-θ-specific inhibitory nucleic acid molecule. 
     
     
         16 . The method of  claim 15 , wherein the vimentin-specific and/or the RLTPR-specific and/or the PCK-θ-specific inhibitory nucleic acid molecule is selected from the group consisting of a RNAi nucleic acid molecule, an antisense nucleic acid molecule, and a siRNA nucleic acid molecule. 
     
     
         17 . The method of  claim 15 , wherein the vimentin-specific inhibitory nucleic acid molecule is complementary to at least a portion of the sequence shown in SEQ ID NO: 1. 
     
     
         18 . The method of  claim 15 , wherein the RLTPR-specific inhibitory nucleic acid molecule is complementary to at least a portion of the sequence shown in SEQ ID NO: 5. 
     
     
         19 . The method of  claim 15 , wherein the PKC-θ-specific inhibitory nucleic acid molecule is complementary to at least a portion of the sequence shown in SEQ ID NO: 9. 
     
     
         20 . The method of  claim 13 , wherein the method is performed in vitro. 
     
     
         21 . The method of  claim 13 , wherein the method is performed in situ. 
     
     
         22 . The method of  claim 21 , wherein the method is performed on an individual who has received or is receiving a bone marrow transplant. 
     
     
         23 . The method of  claim 13 , wherein the Treg cells in which the vimentin, and/or RLTPR and/or PCK-θ has been reduced or eliminated exhibit a phenotype of at least one of the following:
 reduced PKC-θ auto-phosphorylation at Ser676; 
 improved ability to suppress CD4+ and CD8+ Tcon proliferation; 
 increased surface expression of Nrp1; 
 increased surface expression of Lag3; 
 increased basal and maximal oxygen consumption rate (OCR); 
 increased BoDipy C1-C12  uptake; 
 increased expression of CD71; 
 increased expression of CD98; 
 increased expression of CPT1a; or 
 reduced activity of mTORC2, 
 relative to Tregs in which vimentin, RLTPR and/or PCK-θ is not reduced or eliminated.