Methods for altering cell fate to generate T-cells specific for an antigen of interest

• US 20040072288A1
• Filed: 04/30/2003
• Published: 04/15/2004
• Est. Priority Date: 12/22/2000
• Status: Active Grant

• Current Assignee: SAB LLC (W.P. Carey, Inc. (Real Estate))
• Original Assignee: Kyowa Hakko Kirin Company Limited (Kirin Holdings Company Limited)
• Inventors: Robl, James M., Collas, Philippe, Skalhegg, Bjorn Steen
• Granted Patent: US 7,491,534 B2
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/4632 T-cell receptors [TCR]; antibody T-cell receptor constructs
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/464499 Undefined tumor antigens, e.g. tumor lysate or antigens targeted by cells isolated from tumor
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2506/00 Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
• CPC Class Codes: C12N 2517/10 Conditioning of cells for in vitro fecondation or nuclear transfer
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

The invention provides methods for altering the expression profile of a cell to convert the cell from one cell type to a desired cell type (e.g., T-cells). These reprogrammed cells may be used in a variety of medical applications for treating a mammal in need of a particular cell type.

Claims

1. A method of reprogramming a cell to express a T-cell receptor (TCR) reactive with an antigen of interest, said method comprising the steps of:

(a) incubating a nucleus from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said nucleus or the addition of a factor from said reprogramming media to said nucleus;

(b) inserting said nucleus or a chromatin mass formed from said nucleus into a recipient cell or cytoplast, thereby forming a reprogrammed cell; and

(c) selecting said reprogrammed cell that expresses a TCR reactive with an antigen of interest.

7. A cell produced using the method of claim 1, wherein said cell expresses a combination of two or more endogenous mRNA molecules or endogenous proteins that is not expressed in a naturally-occurring cell.

15. The method of claim 1, wherein said antigen is a tumor antigen.

16. The method of claim 1, wherein said antigen is a fat cell antigen.

17. The method of claim 1, wherein said antigen is an antigen from an infectious pathogen, autoimmune generated cell, or autoantibody.

19. The method of claim 1, wherein said nucleus remains membrane-bounded and the chromatin in said nucleus does not condense during incubation with said reprogramming media.

20. The method of claim 1, wherein a chromatin mass is formed from incubation of said nucleus in said reprogramming media.

21. The method of claim 1, wherein said chromatin mass is incubated in an interphase reprogramming media under conditions that allow a nucleus to be formed from said chromatin mass and said reformed nucleus is inserted into said recipient cell or said recipient cytoplast.

23. The method of claim 1, wherein at least 5 mRNA or protein molecules are expressed in said reprogrammed cell that are not expressed in said donor cell or said permeabilized cell.

24. The method of claims 23, wherein said 5 mRNA or protein molecules are specific for T-cells.

25. The method of claim 1, wherein at least 5 mRNA or protein molecules are expressed in said donor cell or said permeabilized cell that are not expressed in said reprogrammed cell.

26. The method of claim 1, wherein said donor cell or said permeabilized cell is an interphase or mitotic cell.

27. The method of claim 1, wherein said donor cell, said permeabilized cell, said recipient cell, said recipient cytoplast, or said reprogrammed cell is an epithelial cell, neural cell, epidermal cell, keratinocyte, hematopoietic cell, melanocyte, chondrocyte, B-cell, Jurak cell, T-cell, erythrocyte, macrophage, monocyte, fibroblast, muscle cell, embryonic stem cell, or adult stem cell.

28. The method of claim 1, wherein said donor cell or said permeabilized cell is a B-cell, Jurak cell, or fibroblast and said reprogrammed cell is a T-cell.

2. A method of reprogramming a cell to express a TCR reactive with an antigen of interest, said method comprising the steps of:

(a) incubating a nucleus from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said nucleus or the addition of a factor from said reprogramming media to said nucleus;

(b) inserting said nucleus or a chromatin mass formed from said nucleus into a recipient cell or cytoplast, thereby forming a reprogrammed cell; and

(c) prior to, during, or after step (a) or (b), inserting into said nucleus, said chromatin mass, or said reprogrammed cell a nucleic acid encoding a TCR reactive with an antigen of interest.

3. A method of reprogramming a cell to express a TCR reactive with an antigen of interest, said method comprising the steps of:

(a) incubating a chromatin mass from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said chromatin mass or the addition of a factor from said reprogramming media to said chromatin mass;

(b) inserting said chromatin mass or a nucleus formed from said chromatin mass into a recipient cell or cytoplast, thereby forming a reprogrammed cell; and

(c) selecting said reprogrammed cell that expresses a TCR reactive with an antigen of interest.

4. A method of reprogramming a cell to express a TCR reactive with an antigen of interest, said method comprising the steps of:

(a) incubating a chromatin mass from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said chromatin mass or the addition of a factor from said reprogramming media to said chromatin mass;

(b) inserting said chromatin mass or a nucleus formed from said chromatin mass into a recipient cell or cytoplast, thereby forming a reprogrammed cell; and

(c) prior to, during, or after step (a) or (b), inserting into said nucleus, said chromatin mass, or said reprogrammed cell a nucleic acid encoding a TCR reactive with an antigen of interest.

5. A method of reprogramming a cell to express a TCR reactive with an antigen of interest, said method comprising the steps of:

(a) incubating a permeabilized cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from the nucleus or chromatin mass of said permeabilized cell or the addition of a factor from said reprogramming media to said nucleus or chromatin mass, thereby forming a reprogrammed cell; and

(b) selecting said reprogrammed cell that expresses a TCR reactive with an antigen of interest.

18. The method of claim 5, wherein said reprogramming media is an interphase reprogramming media or a mitotic reprogramming media.

22. The method of claim 5, wherein said reprogrammed cell is incubated under conditions that allow the membrane of said reprogrammed cell to reseal.

29. The method of claim 5, wherein said recipient cell or said cytoplast is an undifferentiated cell.

30. The method of claim 5, wherein said reprogramming media is a cell extract.

31. The method of claim 5, wherein said reprogramming media comprises RNA or protein for a T-cell receptor reactive with said antigen of interest.

6. A method of reprogramming a cell to express a TCR reactive with an antigen of interest, said method comprising the steps of:

(a) incubating a permeabilized cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from the nucleus or chromatin mass of said permeabilized cell or the addition of a factor from said reprogramming media to said nucleus or chromatin mass, thereby forming a reprogrammed cell; and

(b) prior to, during, or after step (a), inserting into said permeabilized cell or said reprogrammed cell a nucleic acid encoding a TCR reactive with an antigen of interest.

8. A cell that expresses a T-cell receptor reactive with an antigen of interest and one or more fibroblast-specific proteins.

9. A method of treating, stabilizing, or preventing a disease, disorder, or condition in a mammal, said method comprising the steps of:

(a) incubating a nucleus from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said nucleus or the addition of a factor from said reprogramming media to said nucleus;

(b) inserting said nucleus or a chromatin mass formed from said nucleus into a recipient cell or cytoplast, thereby forming a reprogrammed cell;

(c) selecting said reprogrammed cell that expresses a TCR reactive with an antigen of interest; and

(d) administering said reprogrammed cell to a mammal.

32. The method of claim 9, wherein said donor cell, said permeabilized cell, said recipient cell, or said recipient cytoplast is from a human.

33. The method claim 9, wherein said disease, disorder, or condition is an immune disease, autoimmune disease, inflammatory condition, cancer, infection, or obesity.

34. The method of claim 33, wherein said disease, disorder, or condition is cancer.

10. A method of treating, stabilizing, or preventing a disease, disorder, or condition in a mammal, said method comprising the steps of:

(a) incubating a nucleus from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said nucleus or the addition of a factor from said reprogramming media to said nucleus;

(b) inserting said nucleus or a chromatin mass formed from said nucleus into a recipient cell or cytoplast, thereby forming a reprogrammed cell;

(c) prior to, during, or after step (a) or (b), inserting into said nucleus, said chromatin mass, or said reprogrammed cell a nucleic acid encoding a TCR reactive with an antigen of interest; and

(d) administering said reprogrammed cell to a mammal.

11. A method of treating, stabilizing, or preventing a disease, disorder, or condition in a mammal, said method comprising the steps of:

(a) incubating a chromatin mass from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said chromatin mass or the addition of a factor from said reprogramming media to said chromatin mass;

(b) inserting said chromatin mass or a nucleus formed from said chromatin mass into a recipient cell or cytoplast, thereby forming a reprogrammed cell;

(c) selecting said reprogrammed cell that expresses a TCR reactive with an antigen of interest; and

(d) administering said reprogrammed cell to a mammal.

12. A method of treating, stabilizing, or preventing a disease, disorder, or condition in a mammal, said method comprising the steps of:

(a) incubating a chromatin mass from a donor cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from said chromatin mass or the addition of a factor from said reprogramming media to said chromatin mass;

(b) inserting said chromatin mass or a nucleus formed from said chromatin mass into a recipient cell or cytoplast, thereby forming a reprogrammed cell;

(c) prior to, during, or after step (a) or (b), inserting into said nucleus, said chromatin mass, or said reprogrammed cell a nucleic acid encoding a TCR reactive with an antigen of interest; and

(d) administering said reprogrammed cell to a mammal.

13. A method of treating, stabilizing, or preventing a disease, disorder, or condition in a mammal, said method comprising the steps of:

(a) incubating a permeabilized cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from the nucleus or chromatin mass of said permeabilized cell or the addition of a factor from said reprogramming media to said nucleus or chromatin mass, thereby forming a reprogrammed cell;

(b) selecting said reprogrammed cell that expresses a TCR reactive with an antigen of interest; and

(c) administering said reprogrammed cell to a mammal.

14. A method of treating, stabilizing, or preventing a disease, disorder, or condition in a mammal, said method comprising the steps of:

(a) incubating a permeabilized cell with a reprogramming media comprising a T-cell factor under conditions that allow the removal of a factor from the nucleus or chromatin mass of said permeabilized cell or the addition of a factor from said reprogramming media to said nucleus or chromatin mass, thereby forming a reprogrammed cell;

(b) prior to, during, or after step (a), inserting into said nucleus, said chromatin mass, or said reprogrammed cell a nucleic acid encoding a TCR reactive with an antigen of interest; and

(b) administering said reprogrammed cell to a mammal.