Enhanced antigen presenting ability of RNA CAR T cells by co-introduction of costimulatory molecules

Patent metadata

Field	Value
Publication number	US-11028143-B2
Application number	US-201515113044-A
Country	US
Kind code	B2
Filing date	Jan 21, 2015
Priority date	Jan 21, 2014
Publication date	Jun 8, 2021
Grant date	Jun 8, 2021

Abstract

The invention provides T cells comprising nucleic acid sequence encoding a chimeric antigen receptor and a nucleic acid sequence encoding an enhancer of T cell priming, compositions including the T cells, and methods of using the T cells to treat diseases associated with the expression of disease-associated antigens.

First claim

The invention claimed is:

1. A T cell comprising an exogenous nucleic acid, wherein:
(a) the nucleic acid comprises a first nucleic acid sequence encoding a chimeric antigen receptor (CAR) comprising an extracellular domain, a transmembrane domain, and an intracellular signaling domain, wherein the intracellular signaling domain comprises a costimulatory signaling domain that is a functional signaling domain from a protein selected from the group consisting of OX40, CD27, CD28, CD30, CD40, PD-1, CD2, CD7, CD258, NKG2C, B7-H3, ICAM-1, LFA-1 (CD11a/CD18), ICOS and 4-1BB (CD137), or any combination thereof, and
(b) the nucleic acid comprises a second nucleic acid sequence encoding a polypeptide which enhances T cell priming, or a functional fragment or variant thereof, wherein the polypeptide is selected from the group consisting of CD70, CD86, and CD252,
wherein the polypeptide is expressed at a level that: (1) the T cell enhances the priming of another T cell that does not comprise the exogenous nucleic acid and (2) expression of the polypeptide does not inhibit the cell-killing function of the CAR by more than 40%, provided that
(i) the first and/or second nucleic acid sequence comprises an RNA; or
(ii) the CAR further comprises a second intracellular signaling domain.

2. The T cell of claim 1 , wherein:
(i) the first and second nucleic acid sequences are disposed on a single nucleic acid molecule,
(ii) the first and second nucleic acid sequences are disposed on a single nucleic acid molecule, wherein the single nucleic acid molecule comprises an RNA,
(iii) the first and second nucleic acid sequences are disposed on a single nucleic acid molecule, wherein the single nucleic acid molecule comprises a DNA,
(iv) the first and second nucleic acid sequences are disposed on two or more distinct nucleic acid molecules,
(v) the first and second nucleic acid sequences are disposed on two or more distinct nucleic acid molecules, wherein one or both nucleic acid molecules comprise RNA,
(vi) the first and second nucleic acid sequences are disposed on two or more distinct nucleic acid molecules, wherein one or both nucleic acid molecules comprise DNA, or
(vii) the first and second nucleic acid sequences are disposed on two or more distinct nucleic acid molecules, wherein one nucleic acid molecule comprises an RNA and the other nucleic acid molecule comprises a DNA.

3. The T cell of claim 1 , wherein:
(i) the first nucleic acid sequence comprises an RNA,
(ii) the second nucleic acid sequence comprises an RNA, or
(iii) the first and the second nucleic acid sequences each comprise RNA.

4. The T cell of claim 3 , wherein:
(i) the T cell is transfected to transiently express the first and/or second RNAs,
(ii) the cell does not comprise an exogenous DNA encoding the first or second RNA,
(iii) the first and/or second RNAs are generated by in vitro transcription,
(iv) the first and/or second RNAs are synthetic RNAs, or
(v) the first and/or second RNAs are introduced into the T cell by electroporation.

5. The T cell of claim 1 , wherein the first and/or second nucleic acid sequence comprises DNA or cDNA.

6. The T cell of claim 1 , wherein:
(i) the first and/or second nucleic acid sequence comprises a vector,
(ii) the first and/or second nucleic acid sequence comprises a viral vector,
(iii) the first and/or second nucleic acid sequence comprises a retroviral vector or a lentiviral vector, or
(iv) the T cell is virally transduced to express the first and/or second nucleic acid sequence.

7. The T cell of claim 1 , wherein the extracellular domain of the CAR comprises an antigen-binding domain, wherein:
(i) the antigen-binding domain is a scFv domain,
(ii) the antigen-binding domain binds to an antigen associated with a disease state, wherein the disease state is selected from the group consisting of a proliferative disease, a precancerous condition, a non-cancer indication, a viral infection, and a bacterial infection,
(iii) the antigen-binding domain binds to a tumor antigen, a viral antigen, or a bacterial antigen, or
(iv) the antigen-binding domain binds to a tumor antigen, wherein the tumor antigen is an antigen associated with a cancer selected from the group consisting of brain cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer, liver cancer, kidney cancer, lymphoma, leukemia, lung cancer, melanoma, metastatic melanoma, mesothelioma, neuroblastoma, ovarian cancer, prostate cancer, pancreatic cancer, renal cancer, skin cancer, thymoma, sarcoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, uterine cancer, and combinations thereof.

8. The T cell of claim 1 , wherein:
(i) the transmembrane domain comprises a transmembrane protein of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154,
(ii) the intracellular signaling domain comprises a functional signaling domain of CD3 zeta, common FcR gamma (FCER1G), Fc gamma RIIa, FcR beta (Fc Epsilon Rib), CD3 gamma, CD3 delta, CD3 epsilon, CD79a, CD79b, DAP10, or DAP12,
(iii) the second intracellular signaling domain comprises a costimulatory signaling domain,
(iv) the intracellular signaling domain comprises a CD3zeta domain and the second intracellular signaling domain comprises a 4-1BB domain, or
(v) the extracellular domain is connected to the transmembrane domain by a hinge region.

9. The T cell of claim 1 , wherein:
(i) the T cell has enhanced antigen presentation ability relative to a T cell which lacks the second nucleic acid sequence,
(ii) the T cell has increased efficacy in killing tumor cells or reducing tumor size in a subject with a tumor relative to a T cell comprising only the first nucleic acid sequence, or
(iii) the T cell enhances the priming of T cells with a tumor antigen, a viral antigen, a bacterial antigen.

10. The T cell of claim 1 , wherein the T cell is transfected to transiently express a nucleic acid comprising a third nucleic acid sequence encoding a polypeptide which enhances T cell priming, or a functional fragment or variant thereof, which differs from the polypeptide encoded by the second nucleic acid sequence.

11. The T cell of claim 10 , wherein:
(i) the T cell has increased T cell priming ability relative to a T cell comprising the first nucleic acid sequence and second nucleic acid sequence, but not the third nucleic acid sequence,
(ii) the third nucleic acid sequence comprises an RNA,
(iii) the third nucleic acid sequence comprises an RNA, wherein the T cell is transfected to transiently express the third RNA,
(iv) the third nucleic acid sequence comprises an RNA, wherein the cell does not comprise an exogenous DNA encoding the third RNA, or
(v) the CAR comprises one or more costimulatory signaling domains, and the third nucleic acid sequence comprises DNA.

12. The T cell of claim 10 , wherein the T cell further comprises one or more additional distinct nucleic acid sequences encoding a polypeptide which enhances T cell priming, or a functional fragment or variant thereof, which differs from the polypeptides encoded by the second and third nucleic acid sequences.

13. The T cell of claim 12 , wherein:
(i) the one or more additional nucleic acid sequences comprise RNA, or
(ii) the CAR comprises one or more costimulatory signaling domains, and the one or more additional nucleic acids comprise DNA.

14. A composition comprising the T cell of claim 1 .

15. The composition of claim 14 , further comprising a second therapeutic

Assignees

• Novartis Ag
• Univ Pennsylvania

Inventors

• Zhao Yangbing
• Shedlock Devon J
• June Carl H
• Liu Xiaojun

Classifications

• A61K40/4272

  Melan-A/MART · CPC title

• A61K40/4255

  Mesothelin [MSLN] · CPC title

• A61K40/4224

  Molecules with a "CD" designation not provided for elsewhere · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

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