METHODS TO AUGMENT OR ALTER SIGNAL TRANSDUCTION
The technology relates generally to the field of immunology and relates in part to compositions and methods for activating cells, including, for example T cells that express chimeric antigen receptors or recombinant TCRs, and reducing cytotoxicity, using chimeric polypeptides including MyD88 and signaling domains of receptor mediators of costimulation.
1 . A modified cell comprising a nucleic acid that comprises a promoter operably linked to a polynucleotide encoding an inducible chimeric signaling polypeptide, wherein the polypeptide comprises
a) first ligand binding region;
b) a second ligand binding region;
c) a MyD88 polypeptide or a truncated MyD88 polypeptide lacking the TIR domain; and
d) a costimulatory polypeptide cytoplasmic signaling region selected from the group consisting of CD27, CD28, ICOS, 4-1BB, RANK, OX40, CD30, TAC1, CD122, GITR, GITR EEE191RVV, TweakR, BCMA, SOD1, SOD2, SOD3, CD86, C74Ra, Dectin, IL15Ra, ITA4, ITGA5, TCL1A, CTLA4, TIM1, TREMBL2, HVEM, BTN3A1 B30.2 domain, CD40-HCR-CD40, CD40DPP4, RANK88, RANK TRAF6, RANK TRAF2/5, RANK HCR-TRAF2/5, cytoplasmic signaling regions.
2 . The modified cell of claim 1 , wherein the polynucleotide encoding the inducible chimeric signaling polypeptide is a first polynucleotide, and the nucleic acid comprises a second polynucleotide encoding a chimeric antigen receptor.
3 .- 7 . (canceled)
8 . The modified cell claim 1 , wherein the cell is a T cell.
9 . The modified cell claim 1 , wherein the cell is an NK cell.
10 . The modified cell claim 1 , wherein the cell is selected from the group consisting of invariant NK-T cells, gamma delta T cells and tumor infiltrating lymphocytes.
11 .- 21 . (canceled)
22 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of CD28, 4-1BB, OX40, ICOS, BCMA, CD27, CD30, CD122, GITR S180A, GITR EEE191RVV, HVEM, TWEAKR, RANK, RANK TRAF6, RANK TRAF 2/5, RANK HCR-TRAF2/5, CD40-HCR-CD40, SOD1, SOD2, TAC1, SOD3, RANK88, IL15Ra, BTN3A1 (B30.2 domain), CD86, CD74Ra, Dectin, ITA4, ITGA5, TCL1A, CTLA4, TIM1, TREMBL2, CD40-HCR- CD40, and DPP4 polypeptide cytoplasmic signaling regions.
23 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of CD28, 4-1BB, OX40, ICOS, BCMA, CD27, CD30, CD122, GITR S180A, GITR EEE191RVV, HVEM, TWEAKR, RANK, RANK TRAF6, RANK TRAF 2/5, RANK HCR-TRAF2/5, SOD1, SOD2, TAC1, SOD3, RANK88, IL15Ra, BTN3A1 (B30.2 domain), CD86, CD74Ra, Dectin, ITA4, ITGA5, TCL1A, CTLA4, TIM1, TREMBL2, and DPP4 polypeptide cytoplasmic signaling regions.
24 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of CD28, 4-1BB, OX40, ICOS, BCMA, CD27, CD30, CD122, GITR S180A, GITR EEE191RVV, HVEM, TWEAKR, SOD1, SOD2, TAC1, SOD3, IL15Ra, BTN3A1 (B30.2 domain), CD86, CD74Ra, Dectin, ITA4, ITGA5, TCL1A, CTLA4, TIM1, TREMBL2, and DPP4 polypeptide cytoplasmic signaling regions.
25 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of RANK, RANK TRAF6, RANK TRAF 2/5, RANK HCR-TRAF2/5, CD40-HCR-CD40, and RANK88 polypeptide cytoplasmic signaling regions.
26 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of RANK, RANK TRAF6, RANK TRAF 2/5, RANK HCR-TRAF2/5, and RANK88 polypeptide cytoplasmic signaling regions.
27 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of CD27, CD28, ICOS, 4-1BB, RANK/TRANCE-R, OX40, CD30, TweakR, TAC1, BCMA and HVEM polypeptide cytoplasmic signaling regions.
28 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of CD27, CD28, ICOS, OX40, CD30, TweakR, TAC1, BCMA and HVEM polypeptide cytoplasmic signaling regions.
29 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is selected from the group consisting of 4-1BB, RANK, OX40, and HVEM polypeptide cytoplasmic signaling regions.
30 . (canceled)
31 . The modified cell of claim 1 , wherein the costimulatory polypeptide cytoplasmic signaling region is a 4-1BB polypeptide cytoplasmic signaling region.
32 .- 32 . 2 . (canceled)
33 . A pharmaceutical composition comprising the modified cell of claim 1 .
34 . The modified cell of claim 1 , wherein the chimeric signaling polypeptide comprises a membrane targeting region.
35 . The modified cell of claim 34 , wherein the membrane targeting region is selected from the group consisting of a myristoylation region, a palmitoylation region, a prenylation region, and transmembrane sequences of receptors.
36 . The modified cell of claim 34 , wherein the membrane targeting region is a myristoylation region.
37 . The modified cell of claim 36 , wherein the membrane targeting region is a myristoylation region has an amino acid sequence of SEQ ID NO: 4, or an amino acid sequence 95% or more identical to SEQ ID NO:4.
37 . 1 . (canceled)
38 . The modified cell of claim 1 , wherein the inducible chimeric signaling polypeptide comprises two FKBP12 variant polypeptide regions, a truncated MyD88 polypeptide, and a 4-1BB polypeptide cytoplasmic signaling region.
39 . The modified cell of claim 38 , wherein the inducible chimeric signaling polypeptide comprises a myristoylation region.
40 . The modified cell of claim 38 , wherein the inducible chimeric signaling polypeptide comprises no myristoylation region.
41 .- 54 . (canceled)
55 . The modified cell of claim 1 , wherein the first ligand binding region comprises a FKBP12 variant polypeptide region.
56 . The modified cell of claim 1 , wherein the second ligand binding region comprises a FKBP12 variant polypeptide region.
57 . The modified cell of claim 1 , wherein the first ligand binding region and the second ligand binding region each comprises a FKBP12 variant polypeptide region.
58 . The modified cell of claim 1 , wherein the first ligand binding region or the second ligand binding region comprises a FKBP12 wild type polypeptide region.
59 . The modified cell of claim 58 , wherein the first ligand binding region and the second ligand binding region each comprises a FKBP12 wild type polypeptide region.
60 . The modified cell of claim 1 , wherein the first and second ligand binding regions together are Fv′Fvls.
61 . The modified cell of claim 60 , wherein Fv′Fvls comprises two linked polypeptides comprising the amino acid sequence of SEQ ID NO: 6 and the amino acid sequence of SEQ ID NO: 8.
62 .- 66 . (canceled)
67 . The modified cell of claim 1 , wherein the first ligand binding region is an FK506 binding protein 12 (FKBP12) region and the second ligand binding region is an FKBP12-Rapamycin-binding domain of mTOR (FRB) region, or an FRB variant polypeptide region.
68 .- 163 . (canceled)
164 . The modified cell of claim 1 , wherein the inducible chimeric signaling polypeptide comprises no membrane targeting region.