Granted Patent B2

US 12,203,098 · App. 18/345,885 · Granted Jan 21, 2025

Inventors: Bahram Valamehr (San Diego, CA); Tom Tong Lee (San Diego, CA); Martin Hosking (San Diego, CA); Eigen Peralta (San Diego, CA); Chia-Wei Chang (San Diego, CA)

Assignee: FATE THERAPEUTICS, INC.

C12N5/0634A61P35/00C07K14/7051C07K16/3015C07K2317/565C07K2317/622C07K2319/02C07K2319/03

Abstract

Claims (101)

1. A cell or a population thereof, wherein

(i) the cell is (a) an immune cell; (b) an induced pluripotent cell (iPSC); or (c) a derivative effector cell obtained from differentiating the iPSC; and

(ii) the cell comprises a polynucleotide encoding a chimeric antigen receptor (CAR) comprising:

(a) an ectodomain comprising an antigen binding domain recognizing a HER2 (human epidermal growth factor receptor 2) antigen, wherein the antigen binding domain comprises:

(1) a heavy chain variable (VH) domain comprising a heavy chain complementary determining region 1 (H-CDR1) comprising SEQ ID NO: 103, a heavy chain complementary determining region 2 (H-CDR2) comprising SEQ ID NO: 104, and a heavy chain complementary determining region 3 (H-CDR3) comprising SEQ ID NO: 105; and

(2) a light chain variable (VL) domain comprising a light chain complementary determining region 1 (L-CDR1) comprising SEQ ID NO: 106, a light chain complementary determining region 2 (L-CDR2) comprising SEQ ID NO: 107, and a light chain complementary determining region 3 (L-CDR3) comprising SEQ ID NO: 108;

(b) a transmembrane domain; and

(c) an endodomain comprising at least one signaling domain;

wherein the at least one signaling domain responds specifically to binding of the CAR to a HER2 antigen expressed on a cancer cell.

2. The cell or population thereof of claim 1 , wherein the antigen binding domain:

(a) comprises a VH domain with at least 80% sequence identity to SEQ ID NO: 109;

(b) comprises a VL domain with at least 80% sequence identity to SEQ ID NO: 110;

(c) comprises a single chain variable fragment (scFV) comprising VH-linker-VL or VL-linker-VH, wherein the linker varies in length and sequence, and optionally wherein the linker has at least 80% sequence identity to SEQ ID NOs: 111-114;

(d) comprises an scFV represented by an amino acid sequence that is of at least about 99%, about 98%, about 96%, about 95%, about 90%, about 85%, or about 80% identity to SEQ ID NO: 115 or SEQ ID NO: 116, wherein each of SEQ ID NOs: 115 and 116 comprises a linker that varies in length and sequence; and/or

(e) is humanized.

3. The cell or population thereof of claim 1 , wherein the at least one signaling domain comprises:

(a) a signaling domain of any one of: 2B4 (Natural killer Cell Receptor 2B4), 4-1BB (Tumor necrosis factor receptor superfamily member 9), CD16 (IgG Fc region Receptor III-A), CD2 (T-cell surface antigen CD2), CD28 (T-cell-specific surface glycoprotein CD28), CD28H (Transmembrane and immunoglobulin domain-containing protein 2), CD3ζ (T-cell surface glycoprotein CD3 zeta chain), DAP10 (Hematopoietic cell signal transducer), DAP12 (TYRO protein tyrosine kinase-binding protein), DNAM1 (CD226 antigen), FcERIγ (High affinity immunoglobulin epsilon receptor subunit gamma), IL21R (Interleukin-21 receptor), IL-2Rβ/IL15Rβ (Interleukin-2 receptor subunit beta), IL-2Rγ (Cytokine receptor common subunit gamma), IL-7R (Interleukin-7 receptor subunit alpha), KIR2DS2 (Killer cell immunoglobulin-like receptor 2DS2), NKG2D (NKG2-D type II integral membrane protein), NKp30 (Natural cytotoxicity triggering receptor 3), NKp44 (Natural cytotoxicity triggering receptor 2), NKp46 (Natural cytotoxicity triggering receptor 1), CS1 (SLAM family member 7), and CD8 (T-cell surface glycoprotein CD8 alpha chain);

(b) an amino acid sequence that has at least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to the cytoplasmic domain, or a portion thereof, of 2B4, 4-1BB, CD16, CD2, CD28, CD28H, CD3ζ, CD3ζ1XX, DAP10, DAP12, DNAM1, FcERIγ, IL21R, IL2Rβ (IL15Rβ), IL2Rγ, IL7R, KIR2DS2, NKG2D, NKp30, NKp44, NKp46, CS1, or CD8, represented by SEQ ID NOs: 54-76, respectively; and/or

(c) an amino acid sequence that has at least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to the cytoplasmic domain, or a portion thereof, of 2B4, CD28, CD3ζ, DAP10, NKG2D, CD3 ζ1XX, DNAM1, CS1, or combinations thereof.

4. The cell or population thereof of claim 3 , wherein the endodomain comprises two different signaling domains, and wherein said endodomain domain comprises fused cytoplasmic domains, or portions thereof, in any one of the forms: 2B4-CD3ζ/1XX, 2B4-DNAM1, 2B4-FcERIγ, 2B4-DAP10, CD16-DNAM1, CD16-DAP10, CD16-DAP12, CD2-CD3ζ/1XX, CD2-DNAM1, CD2-FcERIγ, CD2-DAP10, CD28-DNAM1, CD28-FcERIγ, CD28-DAP10, CD28-DAP12, CD28-CD3ζ/1XX, CD28H-CD3ζ/1XX, DAP10-CD3 ζ1XX, DAP10-DAP12, DAP12-CD3ζ/1XX, DAP12-DAP10, DNAM1-CD3ζ/1XX, KIR2DS2-CD3ζ/1XX, KIR2DS2-DAP10, KIR2DS2-2B4, or NKp46-2B4.

5. The cell or population thereof of claim 1 , wherein the transmembrane domain comprises an amino acid sequence that has at least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to a transmembrane region, or a portion thereof, of:

(a) CD2, CD3δ, CD3ε, CD3γ, CD3ζ, CD4, CD8, CD8a, CD8b, CD16, CD27, CD28, CD28H, CD40, CD84, CD166, 4-1BB, OX40, ICOS, ICAM-1, CTLA4, PD1, LAG3, 2B4, BTLA, DNAM1, DAP10, DAP12, FcERIγ, IL7, IL12, IL15, KIR2DL4, KIR2DS1, KIR2DS2, NKp30, NKp44, NKp46, NKG2C, NKG2D, CS1, or T cell receptor polypeptide;

(b) 2B4, CD2, CD16, CD28, CD28H, CD3ζ, DAP10, DAP12, DNAM1, FcERIγ, KIR2DS2, NKG2D, NKp30, NKp44, NKp46, CS1, or CD8; or

(c) 2B4, CD28, CD28H, DAP10, DNAM1, KIR2DS2, and NKG2D.

6. The cell or population thereof of claim 1 , wherein the transmembrane domain and its immediately linked signaling domain are from a same protein or from different proteins.

7. The cell or population thereof of claim 1 , wherein the ectodomain comprises one or more of:

(a) a signal peptide; and/or

(b) a spacer/hinge.

8. The cell or population thereof of claim 7 , wherein the spacer/hinge comprises:

(a) an IgG4 spacer, a CD28 spacers, a CD8 spacer, a CH3 spacer, a CH2/CH3 spacer, or any combination thereof;

(b) a short spacer of about 10 to about 80 amino acids; a medium spacer of more than 80 to about 180 amino acids; or a long spacer of more than 180 amino acids; and/or

(c) an amino acid sequence of at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to any of SEQ ID NOs: 96-100.

9. The cell or population thereof of claim 8 , wherein the spacer/hinge comprises a medium spacer, wherein the spacer comprises an amino acid sequence of at least about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to SEQ ID NO: 99.

10. The cell or population thereof of claim 1 , wherein the CAR comprises an amino acid sequence of at least about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% identity to SEQ ID NO: 117.

11. The cell or population thereof of claim 1 , further comprising a solid tumor targeting backbone comprising at least one of:

(a) a polynucleotide encoding a C—X—C motif chemokine receptor or a variant thereof;

(b) a polynucleotide encoding a TGFβ signaling redirector receptor (TGFβ-SRR) comprising a partial or full peptide of the extracellular domain (ECD) of transforming growth factor beta receptor (TGFβR); and

(c) a polynucleotide encoding an allo-immune defense receptor (ADR).

12. The cell or population thereof of claim 11 , wherein the solid tumor targeting backbone further comprises:

(i) CD38 knockout;

(ii) a polynucleotide encoding an exogenous CD16 or a variant thereof; and

(iii) a polynucleotide encoding a cytokine signaling complex comprising a partial or full peptide of a cell surface expressed exogenous cytokine and/or a receptor thereof.

13. The cell or population thereof of claim 1 , wherein the cell further comprises one or more of:

(i) HLA-I deficiency and/or HLA-II deficiency;

(ii) introduction of HLA-G or non-cleavable HLA-G;

(iii) disruption of least one of B2M, CIITA, TAP1, TAP2, Tapasin, NLRC5, RFXANK, RFX5, RFXAP, TCR, NKG2A, NKG2D, CD25, CD44, CD54, CD56, CD58, CD69, CIS, CBL-B, SOCS2, PD1, CTLA4, LAG3, TIM3, and TIGIT;

(iv) introduction of at least one of HLA-E, 4-1BBL, CD3, CD4, CD8, CD47, CD113, CD131, CD137, CD80, PDL1, A 2A R, antigen-specific TCR, chimeric fusion receptor (CFR), Fc receptor, an antibody or functional variant or fragment thereof, a checkpoint inhibitor, an engager, and surface triggering receptor for coupling with an agonist; or

(v) at least one of the genotypes listed in Table 4.

14. The cell or population thereof of claim 11 , wherein the C—X—C motif chemokine receptor comprises CXCR2 or CXCR3.

15. The cell or population thereof of claim 11 , wherein the TGFβ-SRR further comprises a partial or full peptide of the intracellular domain (ICD) of a cytokine receptor comprising IL2R, IL12R, IL18R, IL21R, or any combination thereof.

16. The cell or population thereof of claim 15 , wherein:

(a) the cytokine receptor is IL2Rβ, thereby forming a TGFβR2-IL2Rβ redirector receptor, and the intracellular domain (ICD) of IL2Rβ comprises an amino acid sequence represented by SEQ ID NO: 11; or

(b) the cytokine receptor is IL12Rβ, thereby forming a TGFβR2-IL12Rβ redirector receptor, and the intracellular domain (ICD) of IL12Rβ comprises an amino acid sequence represented by SEQ ID NO: 12 or SEQ ID NO: 13; or

(c) the cytokine receptor is IL18Rβ, thereby forming a TGFβR2-IL18Rβ redirector receptor, and the intracellular domain (ICD) of IL18Rβ comprises an amino acid sequence represented by SEQ ID NO: 14; or

(d) the cytokine receptor is IL21R, thereby forming a TGFβR2-IL21R redirector receptor, and the intracellular domain (ICD) of IL21Rβ comprises an amino acid sequence represented by SEQ ID NO: 15; or

(e) the extracellular domain (ECD) of TGFβR comprises an amino acid sequence represented by SEQ ID NO: 10.

17. The cell or population thereof of claim 15 , wherein the cytokine receptor is a fragment of IL2Rβ, forming a TGFβR2-trIL12Rβ redirector receptor which comprises an amino acid sequence having sequence identity of at least 80%, 85%, 90%, 95%, or 97%, 98%, or 99% to a sequence represented by SEQ ID NO: 16, wherein an amino acid sequence represented by SEQ ID NO: 17 comprised in SEQ ID NO: 16 is variable.

18. The cell or population thereof of claim 11 , wherein the ADR is specific to 4-1BB or to CD38.

19. The cell or population thereof of claim 11 , wherein one or more polynucleotides of the solid tumor targeting backbone are inserted at an endogenous CD38 locus to knock out CD38.

20. The cell or population thereof of claim 12 , wherein the polynucleotide encoding the exogenous CD16 or variant thereof and two or more polynucleotides of the solid tumor targeting backbone are co-expressed in a tri-cistronic construct.

21. The cell or population thereof of claim 12 , wherein the exogenous CD16 or variant thereof comprises at least one of:

(a) a high affinity non-cleavable CD16 (hnCD16);

(b) F176V and S197P in ectodomain domain of CD16;

(c) a full or partial ectodomain originated from CD64;

(d) a non-native (or non-CD16) transmembrane domain;

(e) a non-native (or non-CD16) intracellular domain;

(f) a non-native (or non-CD16) signaling domain;

(g) a non-native stimulatory domain; and

(h) transmembrane, signaling, and stimulatory domains that are not originated from CD16, and are originated from a same or different polypeptide.

22. The cell or population thereof of claim 12 , wherein the cell further comprises the cytokine signaling complex comprising:

(a) a partial or full peptide of a cell surface expressed exogenous cytokine or a receptor thereof comprising at least one of IL2, IL4, IL6, IL7, IL9, IL10, IL11, IL12, IL15, IL18, IL21, or respective receptor thereof; or

(b) at least one of:

(i) co-expression of IL15 and IL15Rα with a self-cleaving peptide in-between;

(ii) a fusion protein of IL15 and IL15Rα;

(iii) an IL15/IL15Rα fusion protein with intracellular domain of IL15Rα truncated;

(iv) a fusion protein of IL15 and membrane bound Sushi domain of IL15Rα;

(v) a fusion protein of IL15 and IL15Rβ;

(vi) a fusion protein of IL15 and common receptor γC, wherein the common receptor γC is native or modified; and

(vii) a homodimer of IL15Rβ;

or

(c) at least one of:

(i) a fusion protein of IL7 and IL7Rα;

(ii) a fusion protein of IL7 and common receptor γC, wherein the common receptor γC is native or modified; and

(iii) a homodimer of IL7Rβ;

and optionally,

(d) is transiently expressed.

23. The cell or population thereof of claim 1 , wherein

(i) the CAR is co-expressed with a cytokine signaling complex in a bicistronic construct, wherein the cytokine signaling complex comprises a partial or full peptide of a cell surface expressed exogenous cytokine and/or a receptor thereof; and/or

(ii) wherein the CAR is inserted at a TCR locus, and optionally is operatively linked to an endogenous promoter of the TCR.

24. The cell or population thereof of claim 23 , wherein

(i) the TCR locus is a constant region of TCR alpha and/or TCR beta; and/or

(ii) the TCR is knocked out by the CAR insertion.

25. The cell or population thereof of claim 1 , wherein the cancer cell is a breast cancer cell, an ovary cancer cell, an endometrium cancer cell, a lung cancer cell, an esophageal cancer cell, a salivary gland cancer cell, a bladder cancer cell, a gastric cancer cell, a colorectal cancer cell, or a head and neck cancer cell.

26. The cell or population thereof of claim 1 , wherein (i) the iPSC is a clonal iPSC, a single cell dissociated iPSC, an iPSC cell line cell, or an iPSC master cell bank (MCB) cell; or (ii) the derivative cell comprises a derivative CD34 + cell, a derivative hematopoietic stem and progenitor cell, a derivative hematopoietic multipotent progenitor cell, a derivative T cell progenitor, a derivative NK cell progenitor, a derivative T lineage cell, a derivative NKT lineage cell, a derivative NK lineage cell, or a derivative B lineage cell; or (iii) the derivative cell comprises a derivative effector cell having one or more functional features that are not present in a counterpart primary T, NK, NKT, and/or B cell.

27. The cell or population thereof of claim 11 , wherein the cell is an NK lineage cell or a T lineage cell, wherein:

(i) the NK lineage cell or the T lineage cell has improved infiltration and/or retention at tumor sites;

(ii) the NK lineage cell is capable of recruiting, and/or migrating T cells to tumor sites; or

(iii) the NK lineage cell or the T lineage cell is capable of reducing tumor immunosuppression in the presence of one or more checkpoint inhibitors.

28. A composition comprising the cell or population thereof of claim 1 .

29. The composition of claim 28 , further comprising one or more therapeutic agents, wherein the one or more therapeutic agents comprise a peptide, a cytokine, a checkpoint inhibitor, a mitogen, a growth factor, a small RNA, a dsRNA (double stranded RNA), mononuclear blood cells, feeder cells, feeder cell components or replacement factors thereof, a vector comprising one or more polynucleic acids of interest, an antibody, a chemotherapeutic agent or a radioactive moiety, or an immunomodulatory drug (IMiD).

30. A method of treating a subject using the composition of claim 28 , the method comprising introducing the composition to a subject in need of an adoptive cell therapy, wherein the subject has a hematological malignancy, a solid tumor, or cancer.

Assignments (1)

Continuity (4)

References Cited (77)