High avidity WT1 T cell receptors and uses thereof

Patent metadata

Field	Value
Publication number	US-12448430-B2
Application number	US-202017438380-A
Country	US
Kind code	B2
Filing date	Mar 10, 2020
Priority date	Mar 11, 2019
Publication date	Oct 21, 2025
Grant date	Oct 21, 2025

Abstract

The present disclosure provides T cell receptors (TCRs) and related binding proteins with high functional avidity against tumor associated antigen p37 from Wilms tumor protein 1 (WT1), T cells expressing such high affinity WT1 specific TCRs, nucleic acids encoding the same, and compositions for use in treating diseases or disorders in which cells overexpress WT1 and/or produce the p37 antigen, such as in cancer.

First claim

What is claimed is:

1. A host cell comprising a heterologous polynucleotide encoding a T cell receptor (TCR), wherein the TCR comprises:
(a) a TCR α-chain variable (Vα) domain comprising the CDR1α amino acid sequence set forth in SEQ ID NO:194, the CDR2a amino acid sequence set forth in SEQ ID NO:195, and the CDR3a amino acid sequence set forth in SEQ ID NO:196 or 12; and
(b) a TCR β-chain variable (VB) domain comprising the CDR1β amino acid sequence set forth in SEQ ID NO:197, the CDR2β amino acid sequence set forth in SEQ ID NO:198, and the CDR3β amino acid sequence set forth in SEQ ID NO:199 or 1,
wherein the TCR binds to a VLDFAPPGA (SEQ ID NO:59): human leukocyte antigen (HLA) complex, wherein the HLA comprises HLA-A*201.

2. The host cell of claim 1 , wherein the TCR specifically binds to the VLDFAPPGA (SEQ ID NO:59): HLA complex.

3. The host cell of claim 1 , wherein the TCR further binds to the VLDFAPPGA (SEQ ID NO:59): human leukocyte antigen (HLA) complex on a cell surface independent of CD8 or in the absence of CD8.

4. The host cell of claim 1 , wherein:
(a) the Vα domain of the TCR comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:253; and/or;
(b) the Vβ domain of the TCR comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO:242.

5. The host cell of claim 1 , wherein the Vα domain of the TCR comprises the amino acid sequence set forth in SEQ ID NO:253, and/or wherein the Vβ domain of the TCR comprises the amino acid sequence set forth in SEQ ID NO:242.

6. The host cell of claim 1 , wherein:
(i) the Vα domain of the TCR consists of the amino acid sequence set forth in SEQ ID NO:253; and/or
(ii) the VB domain of the TCR consists of the amino acid sequence set forth in SEQ ID NO:242.

7. The host cell of claim 1 , wherein the TCR further comprises:
(i) a TCR α-chain constant domain having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:47; and/or
(ii) a TCR β-chain constant domain having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:45 or 46.

8. The host cell of claim 7 , wherein the TCR α-chain constant domain and the TCR β-chain constant domain comprise a non-native cysteine.

9. The host cell of claim 1 , wherein the TCR comprises a TCR α-chain comprising the Vα domain and an α-chain constant domain, wherein
the Vα domain comprises the amino acid sequence set forth in SEQ ID NO: 253, and the α-chain constant domain comprises the amino acid sequence of SEQ ID NO:47.

10. The host cell of claim 1 , wherein the TCR comprises a TCR β-chain comprising the Vβ domain and a β-chain constant domain, wherein the Vβ domain comprises the amino acid sequence set forth in SEQ ID NO:242, and the β-chain constant domain comprises the amino acid sequence of SEQ ID NO:45 or 46.

11. The host cell of claim 1 , wherein the TCR comprises a TCR β-chain comprising the Vβ domain and a β-chain constant domain, wherein:
the Vβ domain consists of the amino acid sequence set forth in SEQ ID NO:242, and the β-chain constant domain consists of the amino acid sequence of SEQ ID NO:45 or 46.

12. The host cell of claim 1 , wherein:
the Vα domain and the Vβ domain of the TCR comprise the amino acid sequences set forth in SEQ ID NOs:253 and 242, respectively.

13. The host cell of claim 12 , wherein the TCR further comprises an α-chain constant domain and/or a β-chain constant domain, wherein the α-chain constant domain comprises the amino acid sequence set forth in SEQ ID NO:47, and wherein the β-chain constant domain comprises the amino acid sequence set forth SEQ ID NO:46.

14. The host cell of claim 1 , wherein the TCR comprises a single-chain TCR (scTCR) or a CAR.

15. The host cell of claim 1 , wherein the heterologous polynucleotide encoding the TCR:
(i) is codon optimized for the host cell;
(ii) has at least 75% identity to or comprises the nucleotide sequence set forth in any one of SEQ ID NOs:64, 75, 86, 97, 108, 109, 110, 111, 122, 133, and 144;
(iii) has at least 95% identity to or comprises the polynucleotide sequence set forth in SEQ ID NO:155; or
(iv) encodes an amino acid sequence having at least 95% identity to or comprising the amino acid sequence set forth in SEQ ID NO:48.

16. The host cell of claim 1 , wherein the heterologous polynucleotide encoding the TCR further comprises:
(i) a polynucleotide encoding a polypeptide that comprises an extracellular portion of a CD8 co-receptor α chain;
ii a polynucleotide encoding a polypeptide that comprises an extracellular portion of a CD8 co-receptor β chain; and/or
(iii) the polynucleotide of (i) and the polynucleotide of (ii); and/or
(iv) the polynucleotide of (i) and the polynucleotide of (ii), and further comprises a polynucleotide encoding a self-cleaving peptide disposed between the polynucleotide of (i) and the polynucleotide of (ii); and/or
(v) the polynucleotide of (i) and the polynucleotide of (ii), and further comprises a polynucleotide encoding a self-cleaving peptide disposed between the polynucleotide encoding the TCR and the polynucleotide of (i) and/or between the polynucleotide encoding the TCR and the polynucleotide of (ii); and/or
(vi) a polynucleotide encoding a self-cleaving peptide disposed between a polynucleotide encoding a TCRα chain of the TCR and a polynucleotide encoding a TCRβ chain of the TCR.

17. The host cell of claim 16 , wherein the heterologous polynucleotide comprises, operably linked in-frame:
(i) (pnCD8α)-(pnSCP 1 )-(pnCD8β)-(pnSCP 2 )-(pnTCR);
(ii) (pnCD8β)-(pnSCP 1 )-(pnCD8α)-(pnSCP 2 )-(pnTCR);
iii) (pnTCR)-(pnSCP 1 )-(pnCD8α)-(pnSCP 2 )-(pnCD8β);
(iv) (pnTCR)-(pnSCP 1 )-(pnCD8β)-(pnSCP 2 )-(pnCD8α);
(v) (pnCD8α)-(pnSCP 1 )-(pnTCR)-(pnSCP 2 )-(pnCD8β); or
(vi) (pnCD8β)-(pnSCP 1 )-(pnTCR)-(pnSCP 2 )-(pnCD8α),
wherein pnCD8a is the polynucleotide encoding the polypeptide that comprises the extracellular portion of the CD8 co-receptor a chain,
wherein pnCD8β is the polynucleotide encoding the polypeptide that comprises the extracellular portion of the CD8 co-receptor chain,
wherein pnTCR is the polynucleotide encoding the TCR,
and wherein pnSCP 1 and pnSCP 2 are each independently a polynucleotide encoding a self-cleaving peptide, wherein the polynucleotides and/or the encoded self-cleaving peptides are the same or different.

18. The host cell of claim 17 , wherein the heterologous polynucleotide comprises, operably linked in-frame:
(i) (pnCD8α)-(pnSCP 1 )-(pnCD8β)-(pnSCP 2 )-(pnTCRβ)-(pnSCP 3 )-(pnTCRα);
(ii) (pnCD8β)-(pnSCP 1 )-(pnCD8α)-(pnSCP 2 )-(pnTCRβ)-(pnSCP 3 )-(pnTCRα);
(iii) (pnCD8α)-(pnSCP 1 )-(pnCD8β)-(pnSCP 2 )-(pnTCRα)-(pnSCP 3 )-(pnTCRβ);
(iv) (pnCD8β)-(pnSCP 1 )-(pnCD8α)-(pnSCP 2 )-(pnTCRα)-(pnSCP 3 )-(pnTCRβ);
(v) (pnTCRβ)-(pnSCP 1 )-(pnTCRα)-(pnSCP 2 )-(pnCD8α)-(pnSCP 3 )-(pnCD8β);
(vi) (pnTCRβ)-(pnSCP 1 )-(pnTCRα)-(pnSCP 2 )-(pnCD8β)-(pnSCP 3 )-(pnCD8α);
(vii) (pnTCRα)-(pnSCP 1 )-(pnTCRβ)-(pnSCP 2 )-(pnCD8α)-(pnSCP 3 )-(pnCD8β); or
(viii) (pnTCRα)-(pnSCP 1 )-(pnTCRβ)-(pnSCP 2 )-(pnCD8β)-(pnSCP 3 )-(pnCD8α),
wherein pnCD8α is the polynucleotide encoding the polypeptide that comprises the extracellular portion of the CD8 co-receptor a chain,
wherein pnCD8β is the polynucleotide encoding the polypeptide that comprises the extracellular portion of the CD8 co-receptor chain,
wherein pnTCRα is the polynucleotide encoding the TCR α chain,
wherein pnTCRβ is the polynucleotide encoding the TCR β chain,
and wherein pnSCP 1

Assignees

• Fred Hutchinson Cancer Center

Inventors

• Schmitt Thomas M
• Chapuis Aude G
• Greenberg Philip D

Classifications

• A61K40/4243

  Wilms tumor 1 [WT1] · CPC title

• A61K40/32

  T-cell receptors [TCR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title

• A61K2239/49

  Breast · CPC title

• A61K2239/54

  Pancreas · CPC title