DIMERIZING AGENT REGULATED IMMUNORECEPTOR COMPLEXES
The present disclosure provides adoptive T cell therapies that have improved DARIC architectures for targeting tumor antigens and recruiting TCR signaling complexes for treating, preventing, or ameliorating at least one symptom of a cancer, infectious disease, autoimmune disease, inflammatory disease, and immunodeficiency, or condition associated therewith.
1 . A non-natural cell comprising:
(a) a first polypeptide comprising: a first binding domain that binds a target antigen expressed on a cancer cell; an FKBP multimerization domain polypeptide or variant thereof; a first transmembrane domain; and a first costimulatory domain; and
(b) a second polypeptide comprising:
(i) a second binding domain that binds to CD3ε, CD3δ or CD3γ; an FRB multimerization domain polypeptide or variant thereof; and a second transmembrane domain; or
(ii) an FRB multimerization domain polypeptide or variant thereof, a linker polypeptide, and a CD3ε, CD3δ or CD3γ polypeptide;
wherein a bridging factor promotes the formation of a polypeptide complex on the non-natural cell surface with the bridging factor associated with and disposed between the multimerization domains of the first and second polypeptides.
2 . The non-natural cell of claim 1 , wherein the FKBP multimerization domain polypeptide is FKBP12.
3 . The non-natural cell of claim 1 , wherein the FRB multimerization domain polypeptide is FRB T2098L or FRB T82L.
4 . The non-natural cell of claim 1 , wherein the bridging factor is selected from the group consisting of: AP21967, sirolimus, everolimus, novolimus, pimecrolimus, ridaforolimus, tacrolimus, temsirolimus, umirolimus, and zotarolimus.
5 . The non-natural cell of claim 1 , wherein the first binding domain comprises an antibody or antigen binding fragment thereof.
6 . (canceled)
7 . The non-natural cell of claim 1 , wherein the first binding domain comprises an scFv.
8 . (canceled)
9 . The non-natural cell of claim 7 , wherein the first binding domain binds a target antigen selected from the group consisting of: alpha folate receptor (FR), α v β 6 integrin, B cell maturation antigen (BCMA), B7-H3 (CD276), B7-H6, carbonic anhydrase IX (CAIX), CD16, CD19, CD20, CD22, CD30, CD33, CD37, CD38, CD44, CD44v6, CD44v7/8, CD70, CD79a, CD79b, CD123, CD133, CD138, CD171, carcinoembryonic antigen (CEA), C-type lectin-like molecule-1 (CLL-1), CD2 subset 1 (CS-1), chondroitin sulfate proteoglycan 4 (CSPG4), cutaneous T cell lymphoma-associated antigen 1 (CTAGE1), epidermal growth factor receptor (EGFR), epidermal growth factor receptor variant III (EGFRvIII), epithelial glycoprotein 2 (EGP2), epithelial glycoprotein 40 (EGP40), epithelial cell adhesion molecule (EPCAM), ephrin type-A receptor 2 (EPHA2), fibroblast activation protein (FAP), Fc Receptor Like 5 (FCRL5), fetal acetylcholinesterase receptor (AchR), ganglioside G2 (GD2), ganglioside G3 (GD3), Glypican-3 (GPC3), EGFR family including ErbB2 (HER2), IL-10Rα, IL-13Rα2, Kappa, cancer/testis antigen 2 (LAGE-1A), Lambda, Lewis-Y (LeY), L1 cell adhesion molecule (L1-CAM), melanoma antigen gene (MAGE)-A1, MAGE-A3, MAGE-A4, MAGE-A6, MAGEA10, melanoma antigen recognized by T cells 1 (MelanA or MART1), Mesothelin (MSLN), MUC1, MUC16, MHC class I chain related proteins A (MICA), MHC class I chain related proteins B (MICB), neural cell adhesion molecule (NCAM), cancer/testis antigen 1 (NY-ESO-1), polysialic acid: placenta-specific 1 (PLAC1), preferentially expressed antigen in melanoma (PRAME), prostate stem cell antigen (PSCA), prostate-specific membrane antigen (PSMA), receptor tyrosine kinase-like orphan receptor 1 (ROR1), synovial sarcoma, X breakpoint 2 (SSX2), Survivin, tumor associated glycoprotein 72 (TAG72), tumor endothelial marker 1 (TEM1/CD248), tumor endothelial marker 7-related (TEM7R), trophoblast glycoprotein (TPBG), UL16-binding protein (ULBP) 1, ULBP2, ULBP3, ULBP4, ULBP5, ULBP6, vascular endothelial growth factor receptor 2 (VEGFR2), and Wilms tumor 1 (WT-1).
10 .- 11 . (canceled)
12 . The non-natural cell of claim 1 , wherein the first transmembrane domain is a CD4 transmembrane domain or a CD8α transmembrane domain.
13 . The non-natural cell of any- claim 1 , wherein the first costimulatory domain is selected from a costimulatory molecule selected from the group consisting of: Toll-like receptor 1 (TLR1), TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, caspase recruitment domain family member 11 (CARD11), CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD94, CD134 (OX40), CD137 (4-1BB), CD278 (ICOS), DNAX-Activation Protein 10 (DAP10), FYN, Linker for activation of T-cells family member 1 (LAT), LCK, SH2 Domain-Containing Leukocyte Protein Of 76 kD (SLP76), T cell receptor associated transmembrane adaptor 1 (TRAT1), TNFR2, TNFRS14, TNFRS18, TNRFS25, and zeta chain of T cell receptor associated protein kinase 70 (ZAP70).
14 .- 16 . (canceled)
17 . The non-natural cell of claim 1 , wherein the second binding domain comprises an antibody or antigen binding fragment thereof that binds CD3ε, CD3δ or CD3γ.
18 .- 21 . (canceled)
22 . The non-natural cell of claim 1 , wherein the second transmembrane domain is a CD8α transmembrane domain or a CD4 transmembrane domain.
23 . The non-natural cell of claim 1 , wherein the second polypeptide further comprises a second costimulatory domain.
24 . The non-natural cell of claim 23 , wherein the costimulatory domain of the second polypeptide is selected from a costimulatory molecule selected from the group consisting of: Toll-like receptor 1 (TLR1), TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, caspase recruitment domain family member 11 (CARD11), CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD94, CD134 (OX40), CD137 (4-1BB), CD278 (ICOS), DNAX-Activation Protein 10 (DAP10), FYN, Linker for activation of T-cells family member 1 (LAT), LCK, SH2 Domain-Containing Leukocyte Protein Of 76 kD (SLP76), T cell receptor associated transmembrane adaptor 1 (TRAT1), TNFR2, TNFRS14, TNFRS18, TNRFS25, and zeta chain of T cell receptor associated protein kinase 70 (ZAP70).
25 . (canceled)
26 . The non-natural cell of claim 1 , further comprising a bridging factor associated with and disposed between the multimerization domains of the first and second polypeptides.
27 .- 75 . (canceled)
76 . A fusion polypeptide comprising:
(a) a first polypeptide comprising: a first binding domain that binds a target antigen expressed on a cancer cell; an FKBP multimerization domain polypeptide or variant thereof; a first transmembrane domain; and a first costimulatory domain;
(b) a polypeptide cleavage signal; and
(c) a second polypeptide comprising:
(i) a second binding domain that binds to CD3ε, CD3δ or CD3γ; an FRB multimerization domain polypeptide or variant thereof; and a second transmembrane domain, or
(ii) an FRB multimerization domain polypeptide or variant thereof, a linker polypeptide, and a CD3ε, CD3δ or CD3γ polypeptide.
77 .- 125 . (canceled)
126 . A polypeptide complex comprising:
(a) a first polypeptide comprising: a first binding domain that binds a target antigen expressed on a cancer cell, an FKBP multimerization domain polypeptide or variant thereof; a first transmembrane domain; and a first costimulatory domain;
(b) a second polypeptide comprising:
(i) a second binding domain that binds to CD3ε, CD3δ or CD3γ; an FRB multimerization domain polypeptide or variant thereof; and a second transmembrane domain, or
(ii) an FRB multimerization domain polypeptide or variant thereof, a linker polypeptide, and a CD3ε, CD3δ or CD3γ polypeptide; and
(c) a bridging factor associated with and disposed between the multimerization domains of the first and second polypeptides.
127 .- 183 . (canceled)
184 . A polynucleotide encoding the first or second polypeptide of claim 1 .
185 .- 186 . (canceled)
187 . A vector comprising the polynucleotide of claim 184 .
188 .- 195 . (canceled)
196 . A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the non-natural cell of claim 1 .
197 . A method of treating a subject in need thereof comprising administering the subject an effective amount of the composition of claim 196 .
198 .- 204 . (canceled)