# US 2020/0399598 A1 · Il-33 secreting immunoresponsive cells and uses thereof

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- Number: US 2020/0399598 A1
- Document ID: US20200399598A1
- Status: Published application
- Kind: A1
- Application number: 15/930,947
- Filed: 2020-05-13
- Published: 2020-12-24
- Earliest claimed: 2017-11-14
- Assignees: Memorial Sloan Kettering Cancer Center
- Applicants: Memorial Sloan-Kettering Cancer Center
- Inventors: Anthony Daniyan; Renier J. Brentjens
- Web page: https://patentgrep.com/patent/US20200399598A1

## CPC

- A61K 38/20: Interleukins [IL]
- A61K 38/177: {Receptors; Cell surface antigens; Cell surface determinants}
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/30: characterised by the recombinant expression of specific molecules in the cells of the immune system
- A61K 40/31: Chimeric antigen receptors [CAR]
- A61K 40/4211: {CD19 or B4}
- C07K 14/4748: {Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE}
- C07K 14/54: Interleukins [IL]
- C07K 14/7051: {T-cell receptor (TcR)-CD3 complex}
- C07K 14/70521: {CD28, CD152}
- C07K 16/2803: {against the immunoglobulin superfamily}
- C12N 5/0636: {T lymphocytes}
- C12N 15/867: Retroviral vectors
- A61K 2039/505: {comprising antibodies}
- A61K 2239/38: characterised by the dose, timing or administration schedule
- A61K 2239/48: Blood cells, e.g. leukemia or lymphoma
- C07K 14/535: Granulocyte CSF; Granulocyte-macrophage CSF
- C07K 14/55: IL-2
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2319/02: containing a signal sequence
- C12N 2501/2333: Interleukin-33 (IL-33)
- C12N 2501/599: with CD designations not provided for elsewhere
- C12N 2510/00: Genetically modified cells
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present disclosure provides methods and compositions for enhancing the immune response toward cancers and pathogens. It relates to an immunoresponsive cell comprising an antigen-recognizing receptor (e.g., a chimeric antigen receptor (CAR) or a T cell receptor (TCR)), and expressing increased level of IL-33. In certain embodiments, the engineered immunoresponsive cells are antigen-directed and have enhanced immune-activating properties.

## Claims

### Claim 1

An immunoresponsive cell comprising:
(a) an antigen-recognizing receptor that binds to an antigen, and
(b) an exogenous IL-33 polypeptide or a fragment thereof.

### Claim 2

An immunoresponsive cell comprising:
(a) an antigen-recognizing receptor that binds to an antigen, and
(b) a modified promoter at an endogenous IL-33 gene locus, wherein the modified promoter enhances gene expression of the endogenous IL-33 gene.

### Claim 3 (depends on 2)

The immunoresponsive cell of claim 2, wherein the modification comprises replacement of an endogenous promoter with a constitutive promoter or an inducible promoter, or insertion of a constitutive promoter or inducible promoter to the promoter region of the endogenous IL-33 gene locus.

### Claim 4 (depends on 3)

The immunoresponsive cell of claim 3, wherein the constitutive promoter is selected from the group consisting of a CMV promoter, an EF1a promoter, a SV40 promoter, a PGK1 promoter, a Ubc promoter, a beta-actin promoter, and a CAG promoter.

### Claim 5 (depends on 3)

The immunoresponsive cell of claim 3, wherein the inducible promoter is selected from the group consisting of a tetracycline response element (TRE) promoter and an estrogen response element (ERE) promoter.

### Claim 6 (depends on 1)

The immunoresponsive cell of claim 1, wherein the antigen is a tumor antigen or a pathogen antigen, optionally wherein the antigen is a tumor antigen.

### Claim 7 (depends on 1)

The immunoresponsive cell of claim 1, wherein the exogenous IL-33 polypeptide is secreted.

### Claim 8 (depends on 1)

The immunoresponsive cell of claim 1, wherein said antigen-recognizing receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR).

### Claim 9 (depends on 1)

The immunoresponsive cell of claim 1, wherein the antigen-recognizing receptor and/or the exogenous IL-33 polypeptide is expressed from a vector.

### Claim 10 (depends on 1)

The immunoresponsive cell of claim 1, wherein the cell is selected from the group consisting of a T cell, a Natural Killer (NK) cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell, a Natural Killer T (NKT) cell, and a pluripotent stem cell from which lymphoid cells may be differentiated.

### Claim 11 (depends on 10)

The immunoresponsive cell of claim 10, wherein said immunoresponsive cell is a T cell.

### Claim 12 (depends on 11)

The immunoresponsive cell of claim 11, wherein said T cell is selected from the group consisting of a cytotoxic T lymphocyte (CTL), a regulatory T cell, a Natural Killer T (NKT) cell, and combinations thereof.

### Claim 13 (depends on 6)

The immunoresponsive cell of claim 6, wherein said antigen is a tumor antigen.

### Claim 14 (depends on 13)

The immunoresponsive cell of claim 13, wherein the tumor antigen is selected from the group consisting of CD19, MUC16, MUC1, CAIX, CEA, CD8, CD7, CD10, CD20, CD22, CD30, CLL1, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD133, CD138, EGP-2, EGP-40, EpCAM, Erb-B2, Erb-B3, erb-B4, FBP, Fetal acetylcholine receptor, folate receptor-a, GD2, GD3, HER-2, hTERT, IL-13R-a2, K-light chain, KDR, LeY, L1 cell adhesion molecule, MAGE-A1, Mesothelin, ERBB2, MAGEA3, p53, MART1, GP100, Proteinase3 (PR1), Tyrosinase, Survivin, hTERT, EphA2, NKG2D ligands, NY-ESO-1, oncofetal antigen (h5T4), PSCA, PSMA, ROR1, TAG-72, VEGF-R2, WT-1, BCMA, CD123, CD44V6, NKCS1, EGF1R, and EGFR-VIII.

### Claim 15 (depends on 14)

The isolated immunoresponsive cell of claim 14, wherein said antigen is CD19.

### Claim 16 (depends on 1)

The immunoresponsive cell of claim 1, wherein said IL-33 polypeptide comprises a heterologous signal sequence at the amino-terminus.

### Claim 17 (depends on 16)

The immunoresponsive cell of claim 16, wherein said heterologous signal sequence is selected from the group consisting of an IL-2 signal sequence, a kappa leader sequence, a CD8 leader sequence, and combinations thereof.

### Claim 18 (depends on 17)

The immunoresponsive cell of claim 17, wherein said heterologous signal sequence is an IL-2 signal sequence.

### Claim 19 (depends on 8)

The immunoresponsive cell of claim 8, wherein the antigen-recognizing receptor is a CAR.

### Claim 20 (depends on 19)

The immunoresponsive cell of claim 19, wherein the CAR comprises an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain.

### Claim 21 (depends on 20)

The immunoresponsive cell of claim 20, wherein the intracellular signaling of the CAR does not comprise a co-stimulatory signaling region.

### Claim 22 (depends on 1)

The immunoresponsive cell of claim 1, wherein the IL-33 peptide comprises (a) an amino acid sequence that is at least about 80% homologous to the sequence set forth in SEQ ID NO: 4 or SEQ ID NO: 21, or (b) the amino acid sequence set forth in SEQ ID NO: 4 or SEQ ID NO: 21.

### Claim 23 (depends on 1)

The immunoresponsive cell of claim 1, wherein the exogenous IL-33 polypeptide enhances an immune response of the immunoresponsive cell, and/or increases anti-tumor cytokine production of the immunoresponsive cell.

### Claim 24 (depends on 23)

The immunoresponsive cell of claim 23, wherein the anti-tumor cytokine is selected from the group consisting of IL-2, GM-CSF and IFN-γ.

### Claim 25 (depends on 1)

A pharmaceutical composition comprising an effective amount of an immunoresponsive cell of claim 1 and a pharmaceutically acceptable excipient.

### Claim 26 (depends on 1)

A method of reducing tumor burden in a subject, and/or treating and/or preventing a neoplasm in a subject, and/or lengthening survival of a subject having a neoplasm, and/or increasing immune-activating cytokine production in response to a tumor antigen or a pathogen antigen in a subject, the method comprising administering to the subject an effective amount of immunoresponsive cells of claim 1 or a pharmaceutical composition comprising thereof.

### Claim 27 (depends on 26)

The method of claim 26, wherein the subject did not receive preconditioning chemotherapy prior to the administration of the cells.

### Claim 28 (depends on 1)

A method for producing an antigen-specific immunoresponsive cell of claim 1, the method comprising introducing into an immunoresponsive cell (a) a first nucleic acid sequence encoding the antigen-recognizing receptor; and (b) a second nucleic sequence encoding the exogenous IL-33.

### Claim 29

A nucleic acid composition comprising (a) a first nucleic acid sequence encoding an antigen-recognizing receptor and (b) a second nucleic acid sequence encoding an exogenous IL-33 polypeptide or a fragment thereof, each optionally operably linked to a promoter element.

### Claim 30 (depends on 29)

A vector comprising the nucleic acid composition of claim 29.

### Claim 31 (depends on 30)

The vector of claim 30, which is a retroviral vector.

### Claim 32 (depends on 1)

A kit comprising an immunoresponsive cell of claim 1.