Methods for expanding SARS-COV2-antigen-specific t cells, compositions and uses related thereto
Provided herein are methods for preparing and characterizing SARS-co2 antigen specific immune cell cultures and preparations and methods of using the same in adoptive immunotherapy for cancer, infections and immune disorders. Also, provided are compositions and methods for generating immune cells expressing synthetic antigen binding receptors targeting SARS-cov2 and methods of use of these cells for the treatment and prevention of COVID-19. Also provided are compositions and methods for determining immune response to SARs-cov2 in a subject, detecting SARS-cov2, measuring cytotoxicity induced by SARS-cov2, and detecting the expression and cytotoxicity of synthetic antigen binding receptors targeting SARS-cov2.
1. A method of generating an adoptive cell therapy composition comprising:
transducing T cells that express a T-cell receptor that specifically binds to a SARS-cov2 encoded first antigen with a nucleic acid encoding at least one synthetic antigen binding receptor (SABR) specific for binding to a second antigen selected from the group consisting of a cell surface marker, a cancer cell-associated antigen, a tumor antigen, an autoimmune-associated antigen, a differentiation antigen, an over-expressed embryonic antigen, a mutated tumor suppressor gene and a tumor antigen resulting from chromosomal translocation.
2. A method of generating an adoptive cell therapy composition comprising: transducing allogeneic or autologous T cells that express a T cell receptor that specifically binds to a SARS-cov2-encoded first antigen presented on a major histocompatibility complex (MHC) with a vector comprising a nucleic acid encoding a synthetic antigen binding receptor (SABR) that specifically binds to a second antigen, wherein the second antigen is a target-cell antigen or a cell surface marker.
3. The method of claim 1 or 2 , wherein said second antigen is selected from the group consisting of CD5; CD19; CD123; CD22; CD30; CD171; CS 1; CD33; epidermal growth factor receptor variant III (EGFRviii); ganglioside G2 (GD2); ganglioside GD3 (aNeu5Ac (2-8) aNeu5Ac (2-3) bDGalp (1-4) bDG1cp (1-1) Cer); TNF receptor family member B cell maturation (BCMA); Tn antigen; prostate-specific membrane antigen (PSMA); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Fms Like Tyrosine Kinase 3 (FL T3); Tumor-associated glycoprotein 72 (TAG72); CD38; CD44v6; a glycosylated CD43 epitope expressed on acute leukemia or lymphoma but not on hematopoietic progenitors; a glycosylated CD43 epitope expressed on nonhematopoietic cancers; Carcinoembryonic antigen (CEA); Epithelial cell adhesion molecule (EPCAM); B7H3 (CD276); KIT (CD117); Interleukin-13 receptor subunit alpha-2; Mesothelin; Interleukin 11 receptor alpha (IL-11Ra); prostate stem cell antigen (PSCA); Protease Serine 21; vascular endothelial growth factor receptor 2 (VEGFR2); Lewis (Y) antigen; CD24; Platelet-derived growth factor receptor beta (PDGFR-beta); Stage-specific embryonic antigen-4 (SSEA-4); CD20; Folate receptor alpha; Folate receptor beta (FRb); Receptor tyrosine-protein kinase ERBB2; Mucin 1, cell surface associated (MUC1); epidermal growth factor receptor (EGFR); neural cell adhesion molecule (NCAM); Prostase; prostatic acid phosphatase (PAP); elongation factor 2 mutated (ELF2M); Ephrin 82; fibroblast activation protein alpha; insulin-like growth factor 1 receptor (IGF-1 receptor); carbonic anhydrase IX (CAIX); Proteasome Subunit, Beta Type, 9; glycoprotein 100 (gp100); oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcrabl); tyrosinase; ephrin type-A receptor 2 (EphA2); sialyl Lewis adhesion molecule (sle); ganglioside GM3 (aNeuSAc (2-3) bDClalpd-4) bDGlcp (-1) Cer); transglutaminase 5 (TGS5); high molecular weight-melanoma associated antigen (HMWMAA); o-acetyl-GD2 ganglioside (OAcGD2); tumor endothelial marker 1 (TEM 1/CD248); tumor endothelial marker 7-related (TEM7R); claudin 6 (CLDN6); thyroid stimulating hormone receptor (TSHR); G protein coupled receptor class C group 5, member D (GPRC5D); chromosome X open reading frame 61 (CXORF61); CD97; CD179a; anaplastic lymphoma kinase (ALK); Polysialic acid; placenta-specific 1 (PLAC1); hexasaccharide portion of globoH glycoceramide (GloboH); mammary gland differentiation antigen NY-BR-1 (NY-BR-1); uroplakin 2 (UPK2); adrenoceptor beta 3 (ADRB3); pannexin 3 (PANX3); G protein-coupled receptor 20 (GPR20); lymphocyte antigen 6 complex, locus K 9 (L Y6K); Olfactory receptor 51 E2 (OR51 E2); TCR Gamma Alternate Reading Frame Protein (TARP); Wilms tumor protein (WT1); Cancer/testis antigen 1; Cancer/testis antigen 2; Melanoma-associated antigen 1 (MAGE-A 1); ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML); sperm protein 17 (SPA17); X Antigen Family, Member IA; angiopoietin-binding cell surface receptor 2 (Tie 2); melanoma cancer testis antigen-1; melanoma cancer testis antigen-2; Fos-related antigen 1; tumor protein p53 (p53); p53 mutant; prostein; survivin; telomerase; prostate carcinoma tumor antigen-1; melanoma antigen recognized by T cells 1; Rat sarcoma (Ras) mutant; human Telomerase reverse transcriptase (hTERT); sarcoma translocation breakpoints; melanoma inhibitor of apoptosis (ML-IAP); transmembrane protease, serine 2 (TMPRSS2) ETS fusion gene; N-Acetyl glucosaminyl-transferase V; paired box protein Pax-3 (PAX3); Androgen receptor; Cyclin B1; v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN); Ras Homolog Family Member C (RhoC); Tyrosinase-related protein 2 (TRP-2); Cytochrome P450 1B 1 (CYPIB 1); CCCTC-Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3); Paired box protein Pax-5 (PAX5); proacrosin binding protein sp32; lymphocyte-specific protein tyrosine kinase (LCK); A kinase anchor protein 4 (AKAP-4); synovial sarcoma, X breakpoint 2 (SSX2); Receptor for Advanced Glycation Endproducts; renal ubiquitous 1 (RU1); renal ubiquitous 2 (RU2); legumain; intestinal carboxyl esterase; heat shock protein 70-2 mutated; CD79a; CD79b; CD72; Leukocyte-associated immunoglobulin-like receptor 1 (LAI R 1; Fc fragment of IgA receptor; Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); CD300 molecule-like family member f (CD300LF); C-type lectin domain family 12 member A (CLEC12A); bone marrow stromal cell antigen 2 (BST2); EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2); lymphocyte antigen 75 (LY75); Glypican-3 (GPC3); Fc receptor-like 5 (FCRL5); immunoglobulin lambda-like polypeptide 1 (IGLL 1), MPL; Biotin; c-MYC epitope Tag; CD34; LAMP1; TROP2; GFRalpha4; CDH17; CDH6; NYBR1; CDH19; CD200R; Slea; Fucosyl-GM1; PTK7; gpNMB; CDH1-CD324; DLL3; CD276/B7H3; IL 11 Ra; IL 13Ra2; CD179b-IGLII; TCR-gamma-delta; NKG2D; CD32; Tim1-/HVCR1; CSF2RA; TGFbetaR2; Lews Ag, TCR-beta1 chain; TCR-beta2 chain; TCR-gamma chain; TCR-delta chain; FITC; Luteinizing hormone receptor (LHR); Follicle stimulating hormone receptor (FSHR); Gonadotropin Hormone receptor; CCR4; GD3; SLAMF6; SLAMF4; HIV1 envelope glycoprotein; HTLV1-Tax; CMV pp65; EBVEBNA3c; KSHV K8.1; KSHV-gH; GAD; PDL 1; Guanylyl cyclase C (GCC); auto antibody to desmoglein 3 (Dsg3); auto antibody to desmoglein 1 (Dsg1); HLA, HLA-A; HLA-A2; HLA-B; HLA-C; HLA-DP; HLA-DM; HLA-DOA; HLA-008HLA-DOB; HLA-DQ; HLA-DR; HLA-G; IgE; CD99; Ras G12V; Tissue Factor 1 (TF1); AFP; GPRC5D; Claudin18.2; P-glycoprotein; STEAP1; Liv1; Nectin-4; Cripto; gpA33; BST1/CD157; low conductance chloride channel; the antigen recognized by TNT antibody; CD229; Toso; and BAFF-R.
4. The method of claim 1 , wherein the nucleic acid encoding the SABR is introduced into T cells using a vector, wherein optionally the vector is a retroviral vector, a lentiviral vector, or an in vitro transcribed RNA vector.
5. The method of claim 1 , wherein SARS-cov2 is the etiological agent of coronavirus disease 2019 (COVID-19) or a variant thereof.
6. The method of claim 1 , wherein the SARS-cov2 peptide antigen is comprised of at least 5 but no more than 25 contiguous amino acids sequence of any SARS-cov2 viral protein or a variant thereof.
7. The method of claim 6 , wherein the SARS-cov2 peptide antigen is selected from one or more of the following:
a) a sequence of at least 5 but no more than 25 contiguous amino acids derived from one or more of SEQ ID NO: 694, 700-703, 707, or 712, or a variant with at least 80% sequence homology thereof; and/or
b) a sequence with at least 80% homology to a peptide sequence selected from SEQ ID NO: 980-1028; and/or
c) a sequence with up to 3 conservative substitutions in a peptide sequence selected from SEQ ID NO: 980-1028.
8. The method of claim 1 , wherein the T cells are obtained from a SARS-cov2-immune or a SARS-cov2-positive subject.
9. The method of claim 8 , wherein the SARS-cov2-immune or a SARS-cov2-positive subject has one or more of the following characteristics:
(a) evidence of cellular and/or humoral immunity to SARS-cov2;
(b) a history of infection with SARS-cov2;
(c) history of having received a SARS-cov2 vaccine;
(d) history of receiving one or more SARS-cov2 vaccine booster dose;
(e) T cells that recognize one or more antigens encoded by SARS-cov2 wherein the level of the SARS-cov2 specific T cells is at least 20% higher than a subject who is not SARS-cov2 immune and/or is in the top 20, 30, 40, 50, 60, 70, 80 or 90 percentile of the SARS-cov2 immune subjects; and/or
(f) antibodies against one or more antigens encoded by SARS-cov2; wherein the level of the SARS-cov2 antibodies is at least 20% higher than a subject who is not SARS-cov2 immune and/or is in the top 20, 30, 40, 50, 60, 70, 80 or 90 percentile of the SARS-cov2 immune subjects.
10. The method of claim 1 , wherein the T cells are selected from the group consisting of CD8+ T cells, CD4+ T cells, TREGs, cytotoxic T cells, NKT cells, naïve T cells, central memory T cells, effector memory T cells, stem memory T cells (Tscm), iPSC-derived T cells and synthetic T cells.
11. The method of claim 1 , wherein the T cells have one or more of the features selected from the group consisting of:
(a) lacks the expression of one or more of endogenous T cell receptor chains;
(b) lacks the expression of constant chains of endogenous T cell receptor α, β1, β2, pre-TCRα, γ or δ or combination thereof;
(c) lacks the expression of HLA;
(d) lacks the expression of β2 microglobulin;
(e) lacks the expression of CD52;
(f) expresses an agent which enhances the activity of a SABR-expressing cell;
(g) expresses an agent that provides co-stimulation to a SABR-expressing cell;
(h) expresses a cytokine or a chemokine;
(i) expresses an agent that that promotes the proliferation, persistence, expansion and/or activation of a SABR-expressing cell;
(i) expresses an agent which increases the expression and/or activity of SABR chains;
(k) expresses a soluble receptor;
(l) expresses a second SABR;
(m) expresses an inhibitory SABR that comprises an antigen binding domain, a transmembrane domain and an intracellular domain of an inhibitory molecule;
(n) expresses a second SABR comprising an antigen binding domain, a transmembrane domain and a primary signaling domain;
(o) expresses a second SABR comprising an antigen binding domain, a transmembrane domain, a costimulatory signaling domain but no primary signaling domain;
(p) expresses a second SABR comprising an antigen binding domain, a transmembrane domain, a primary signaling domain and a costimulatory domain;
(q) expresses an accessory module which increases, decreases, regulates or modifies the expression or activity of a SABR or a SABR-expressing cell, wherein the accessory module is selected from the group consisting of POL 1, PDL2, CD80, CD86, NEMO-K277A, K13-opt, MyD88, 41BBL, vFLIPK13, IL-7, IL-15, IL-21, and any combination thereof; and/or
(r) expresses a therapeutic control selected from the group consisting of CNB30, a truncated epidermal growth factor receptor (tEGFR), truncated epidermal growth factor receptor viii (tEGFRviii), truncated CD 19 (tCD 19), truncated BCMA (tBCMA), icaspase-9 and an antibiotic resistance cassette.
12. The method of claim 1 , wherein the SABR comprises:
(a) at least one antigen binding domain and at least one transmembrane or a membrane anchoring domain; and/or
(b) a targeting/binding moiety that is associated with one or more signaling domains.
13. The method of claim 12 , wherein the antigen binding domain and/or the targeting moiety of the SABR comprises a member selected from the group consisting of a scFv, a vL, a vH, an antibody, an antibody fragment, an antibody like moiety, a Vα domain, a Vβ domain, a cytokine, a receptor and combination thereof.
14. The method of claim 1 , wherein the SABR is selected from the group consisting of a Chimeric Antigen Receptor (CAR), an Antibody-TCR chimeric molecule (Ab-TCR), a TCR receptor fusion protein (TFP), a Synthetic Immune Receptor (SIR), a Tri-functional T cell antigen coupler (Tri-TAC or TAC), a z-SIR, a TRUCK, a Universal SABR, a Self-driving SABR, an Armored SABR, a Self-destruct SABR, a Conditional SABR, a Marked SABR, a Tandem SABR (tanSABR), a Dual SABR, a bispecific-SABR, and a safety SABR (sSABR).
15. The method of claim 1 , further comprising a step wherein the T cells are stimulated with antigen presenting cells (APC) presenting SARS-cov2 encoded peptide antigens in vitro and/or in vivo: (a) before transduction with the nucleic acid encoding the SABR; (b) after transduction with the nucleic acid encoding the SABR; or (c) both before and after transduction with the nucleic acid encoding the SABR.
16. A method for generating a composition for the treatment of cancer and/or autoimmune disorders comprising:
transducing T cells that express a T-cell receptor that specifically binds to a SARS-cov2 encoded first antigen with a vector comprising a nucleic acid encoding at least one synthetic antigen binding receptor (SABR) specific for binding to a second antigen selected from the group consisting of CD5; CD19; CD123; CD22; CD30; CD171; CS1; C-type lectin-like molecule-1; CD33; epidermal growth factor receptor variant III (EGFRviii); ganglioside G2 (GD2); ganglioside GD3 (aNeu5Ac (2-8) aNeu5Ac (2-3) bDGalp (1-4) bDG1cp (1-1) Cer); TNF receptor family member B cell maturation (BCMA); Tn antigen; prostate-specific membrane antigen (PSMA); Receptor tyrosine kinase-like orphan receptor 1 (ROR1); Fms Like Tyrosine Kinase 3 (FLT3); Tumor-associated glycoprotein 72 (TAG72); CD38; CD44v6; a glycosylated CD43 epitope expressed on acute leukemia or lymphoma but not on hematopoietic progenitors; a glycosylated CD43 epitope expressed on nonhematopoietic cancers; Carcinoembryonic antigen (CEA); Epithelial cell adhesion molecule (EPCAM); B7H3 (CD276); KIT (CD117); Interleukin-13 receptor subunit alpha-2; Mesothelin; Interleukin 11 receptor alpha (IL-11Ra); prostate stem cell antigen (PSCA); Protease Serine 21; vascular endothelial growth factor receptor 2 (VEGFR2); Lewis (Y) antigen; CD24; Platelet-derived growth factor receptor beta (PDGFR-beta); Stage-specific embryonic antigen-4 (SSEA-4); CD20; Folate receptor alpha; Folate receptor beta (FRb); Receptor tyrosine-protein kinase ERBB2; Mucin 1, cell surface associated (MUC1); epidermal growth factor receptor (EGFR); neural cell adhesion molecule (NCAM); Prostase; prostatic acid phosphatase (PAP); elongation factor 2 mutated (ELF2M); Ephrin 82; fibroblast activation protein alpha; insulin-like growth factor 1 receptor (IGF-1 receptor); carbonic anhydrase IX (CAIX); Proteasome Subunit, Beta Type, 9; glycoprotein 100 (gp100); oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcrabl); tyrosinase; ephrin type-A receptor 2 (EphA2); sialyl Lewis adhesion molecule (sLe); ganglioside GM3 (aNeuSAc (2-3) bDClalpd-4) bDGlcp(-1) Cer); transglutaminase 5 (TGS5); high molecular weight-melanoma associated antigen (HMWMAA); o-acetyl-GD2 ganglioside (OAcGD2); tumor endothelial marker 1 (TEM 1/CD248); tumor endothelial marker 7-related (TEM7R); claudin 6 (CLDN6); thyroid stimulating hormone receptor (TSHR); G protein coupled receptor class C group 5, member D (GPRC5D); chromosome X open reading frame 61 (CXORF61); CD97; CD179a; anaplastic lymphoma kinase (ALK); Polysialic acid; placenta-specific 1 (PLAC1); hexasaccharide portion of globoH glycoceramide (GloboH); mammary gland differentiation antigen NY-BR-1; uroplakin 2 (UPK2); adrenoceptor beta 3 (ADRB3); pannexin 3 (PANX3); G protein-coupled receptor 20 (GPR20); lymphocyte antigen 6 complex, locus K 9 (L Y6K); Olfactory receptor 51E2 (OR51E2); TCR Gamma Alternate Reading Frame Protein (TARP); Wilms tumor protein (WT1); Cancer/testis antigen 1 (NY-ESO-1); Cancer/testis antigen 2; Melanoma-associated antigen 1 (MAGE-A1); ETS translocation-variant gene 6, located on chromosome 12p (ETV6-AML); sperm protein 17 (SPA17); X Antigen Family, Member IA (XAGEI); angiopoietin-binding cell surface receptor 2 (Tie 2); melanoma cancer testis antigen-1; melanoma cancer testis antigen-2; Fos-related antigen 1; tumor protein p53 (p53); p53 mutant; prostein; survivin; telomerase; prostate carcinoma tumor antigen-1; melanoma antigen recognized by T cells 1; Rat sarcoma (Ras) mutant; human Telomerase reverse transcriptase (hTERT); sarcoma translocation breakpoints; melanoma inhibitor of apoptosis (ML-IAP); transmembrane protease, serine 2 ETS fusion gene; N-Acetyl glucosaminyl-transferase V; paired box protein Pax-3 (PAX3); Androgen receptor; Cyclin B1; v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN); Ras Homolog Family Member C (RhoC); Tyrosinase-related protein 2 (TRP-2); Cytochrome P450 1B 1 (CYPIB 1); CCCTC-Binding Factor (Zinc Finger Protein)-Like, Squamous Cell Carcinoma Antigen Recognized By T Cells 3 (SART3); Paired box protein Pax-5 (PAX5); proacrosin binding protein sp32; lymphocyte-specific protein tyrosine kinase (LCK); A kinase anchor protein 4 (AKAP-4); synovial sarcoma, X breakpoint 2 (SSX2); Receptor for Advanced Glycation Endproducts; renal ubiquitous 1 (RU1); renal ubiquitous 2 (RU2); legumain; intestinal carboxyl esterase; heat shock protein 70-2 mutated (mut hsp70-2); CD79a; CD79b; CD72; Leukocyte-associated immunoglobulin-like receptor 1 (LAI R 1; Fc fragment of IgA receptor; Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); CD300 molecule-like family member f (CD300LF); C-type lectin domain family 12 member A (CLEC12A); bone marrow stromal cell antigen 2 (BST2); EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2); lymphocyte antigen 75 (LY75); Glypican-3 (GPC3); Fc receptor-like 5 (FCRL5); immunoglobulin lambda-like polypeptide 1 (IGLL 1), MPL; Biotin; c-MYC epitope Tag; CD34; LAMP1; TROP2; GFRalpha4; CDH17; CDH6; NYBR1; CDH19; CD200R; Slea; Fucosyl-GM1; PTK7; gpNMB; CDH1-CD324; DLL3; CD276/B7H3; IL 11 Ra; IL 13Ra2; CD179b-IGLII; TCR-gamma-delta; NKG2D; CD32; Tim1-/HVCR1; CSF2RA; TGFbetaR2; TCR-beta1 chain; TCR-beta2 chain; TCR-gamma chain; TCR-delta chain; FITC; Luteinizing hormone receptor (LHR); Follicle stimulating hormone receptor (FSHR); Gonadotropin Hormone receptor; CCR4; GD3; SLAMF6; SLAMF4; HIV1 envelope glycoprotein; HTLV1-Tax; CMV pp65; EBVEBNA3c; KSHV K8.1; KSHV-gH; GAD; PDL1; Guanylyl cyclase C (GCC); auto antibody to desmoglein 3 (Dsg3); auto antibody to desmoglein 1 (Dsg1); HLA-A; HLA-A2; HLA-B; HLA-C; HLA-DP; HLA-DM; HLA-DOA; HLA-DOB; HLA-DQ; HLA-DR; HLA-G; IgE; CD99; Ras G12V; Tissue Factor 1 (TF1); AFP; GPRC5D; Claudin18.2; P-glycoprotein; STEAP1; Liv1; Nectin-4; Cripto; gpA33; BST1/CD157; low conductance chloride channel; the antigen recognized by TNT antibody; CD229; Toso; and BAFF-R;
wherein the method does not comprise the expansion of the starting population of T cells in the presence of SARS-cov2 specific peptides or peptide libraries.
17. A composition for the treatment of cancer and/or autoimmune disorders generated by the method of claim 1 or 16 .
18. A method of treatment of cancer and/or autoimmune disorders comprising administering to a subject in need thereof an effective amount of a composition of claim 17 .