CAL-T constructs and uses thereof
The technology described herein is directed to compositions comprising components of multi-component CALs or CARs, e.g., a TCR recognition domain; and one or both of: (a) an intracellular signaling domain; and (b) a first-type protein interaction domain. Further provided herein are methods for treating or preventing an autoimmune disease, a transplant rejection, or graft versus host disease.
1. A composition, comprising:
a) a soluble extracellular molecule or a soluble extracellular complex comprising:
a TCR recognition domain that binds specifically to a variable region of a TCR on target cells, and a first biomolecular interaction domain of a binding pair of biomolecular interaction domains; and
b) an engineered cell expressing or comprising an engineered signaling polypeptide comprising:
an extracellular second biomolecular interaction domain of the binding pair of biomolecular interaction domains, and an intracellular signaling domain,
wherein the engineered cell is a T-cell or an NK cell or a dendritic cell, wherein the composition is characterized in that it exhibits cell killing activity against a population of the target cells expressing the variable region of the TCR.
2. The composition of claim 1 , wherein the TCR recognition domain comprises a MHC (Major Histocompatibility Complex); a MI-IC-peptide complex; a featureless peptide MHC; or a MHC-peptide fusion.
3. The composition of claim 1 , wherein the TCR recognition domain is a monomer, a dimer, a trimer, a tetramer, a pentamer, a dextramer, or an other oligomer form.
4. The composition of claim 1 ,
a) wherein the binding pair of biomolecular interaction domains are collectively a pair of leucine zippers;
b) wherein the binding pair of biomolecular interaction domains are collectively a BZip (RR) and a AZip (EE);
c) wherein the binding pair of biomolecular interaction domains is a PSD95-Dlg1-zo-1 (PDZ) domain;
d) wherein the binding pair of biomolecular interaction domains are collectively a streptavidin and a streptavidin binding protein (SBP);
e) wherein the binding pair of biomolecular interaction domains are collectively a FKBP-binding domain of mTOR (FRB) and a FK506 binding protein (FKBP);
f) wherein the binding pair of biomolecular interaction domains are collectively a cyclophilin-Fas fusion protein (CyP-Fas) and a FK506 binding protein (FKBP);
g) wherein the binding pair of biomolecular interaction domains are collectively a calcineurin A (CNA) and a FK506 binding protein (FKBP);
h) wherein the binding pair of biomolecular interaction domains are collectively a gibberellin insensitive (GIA) and a gibberellin insensitive dwarf1 (GID1);
i) wherein the binding pair of biomolecular interaction domains are collectively a Snap-tag and a Halo tag;
j) wherein the binding pair of biomolecular interaction domains are collectively a T14-3-3-cdeltaC and a C-Terminal peptides of PMA2 (CT52);
k) wherein the binding pair of biomolecular interaction domains are collectively a PYL and a ABI;
l) wherein the binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain;
m) wherein the binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags;
o) wherein the binding pair of protein interaction domains are collectively a FITC and a FITC binding protein; and/or
p) wherein the binding pair of protein interaction domains are collectively a (R)-Phycoerythrin (R-PE/PE) and a R-PE/PE binding protein.
5. The composition of claim 1 , wherein
a) the binding pair of biomolecular interaction domains are collectively a nucleotide tag and a zinc finger domain and the nucleotide tag is a DNA tag or a dsDNA tag: or
b) wherein the binding pair of biomolecular interaction domains are collectively a pair of nucleotide tags and the nucleotide tags are a DNA tags or a dsDNA tags.
6. The composition of claim 1 , wherein the TCR recognition domain is allogeneic, autologous, or xenogenic to the engineered cell.
7. The composition of claim 1 , wherein the TCR recognition domain is synthetic.
8. The composition of claim 1 , wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is allogeneic, autologous, or xenogenic to the engineered cell.
9. The composition of claim 1 , wherein the TCR recognition domain comprises a MHC and a peptide, wherein the peptide is synthetic.
10. The composition of claim 1 , wherein the engineered cell is a NK cell, a dendritic cell, a regulatory T cell, or an effector T cell.
11. The composition of claim 1 , wherein the engineered cell is further engineered to knockdown the native MHCI/II expressed on the cell surface.
12. The composition of claim 1 , wherein the engineered cell is a NK cell.
13. The composition of claim 1 , wherein the cell further comprises a TCR signaling-responsive promoter operatively linked to a payload transgene.
14. The composition of claim 13 , wherein the TCR signaling-responsive promoter is a NFAT-sensitive promoter.
15. The composition of claim 13 , wherein the payload transgene encodes a checkpoint inhibitor or a proinflammatory cytokine.
16. The composition of claim 15 , wherein the checkpoint inhibitor is ipilimumab, tremelimumab, nivolumab, pembrolizumab, or pidilizumab.
17. The composition of claim 15 , wherein the proinflammatory cytokine is IFN-γ, TNFα, TGF-β, IL-1β, IL-6, IL-4, IL-10, IL-13, IL-2, IL-12, IL-15, or IL-27.