METHODS AND COMPOSITIONS FOR INCREASING THE SUPPRESSIVE FUNCTION OF REGULATORY T-CELLS (TREGS)
Methods and compositions for increasing the suppressive function of regulatory T-cells (Tregs) are provided.
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.