CD8a and t cell receptor variants and methods of using same in modulating immune cell responses

Patent metadata

Field	Value
Publication number	US-10975137-B2
Application number	US-201515524887-A
Country	US
Kind code	B2
Filing date	Nov 6, 2015
Priority date	Nov 6, 2014
Publication date	Apr 13, 2021
Grant date	Apr 13, 2021

Abstract

Novel costimulatory fusion proteins and DNA sequences that enhance T cell responses to weakly immunogenic and/or lowly expressed antigens and that confer T cell resistance against MDSC-mediated suppression are disclosed. The fusion proteins comprise portions of CD4, CD8α or the T cell receptor linked to a specific region of MyD88 or other signaling molecules. These fusion proteins and sequence variants thereof improve T cell activation and responsiveness. Also disclosed is the use of these molecules in host cells as a means to enhance and costimulate responses of immune cells including cytotoxic CD8+ T cells and the use of these cells to treat cancer, infectious agents and other diseases.

First claim

What is claimed is:

1. A fusion protein comprising an amino-terminal domain linked to a region of MyD88 lacking the TIR domain,
wherein the amino-terminal domain is selected from the group consisting of:
(a) extracellular and transmembrane regions of CD8α,
(b) a transmembrane region of CD8α,
(c) extracellular and transmembrane regions of CD4, and
(d) a T cell receptor; and
wherein the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24) plus or minus up to 10 amino acids from either end or from both ends.

2. The fusion protein of claim 1 , wherein (a) the extracellular and transmembrane regions of CD8α correspond to amino acids 1-217 of mouse CD8α (SEQ ID NO:16) or amino acids 1-203 of human CD8α (SEQ ID NO:12) and (b) the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).

3. The fusion protein of claim 1 , wherein the fusion protein comprises mCD8α-hMyD88 as set forth in SEQ ID NO:17 or hCD8α-hMyD88 as set forth in SEQ ID NO:14.

4. The fusion protein of claim 1 , wherein the transmembrane region of CD8α corresponds to amino acids 1-83 of SEQ ID NO:18 and the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).

5. The fusion protein of claim 1 , wherein the fusion protein comprises hCD8αTM-hMyD88 as set forth in SEQ ID NO:18.

6. The fusion protein of claim 1 , wherein (a) the extracellular and transmembrane regions of CD4 correspond to amino acids 1-417 of mouse CD4 (amino acids 1-417 of SEQ ID NO:21) or human CD4 (amino acids 1-418 of SEQ ID NO:20) and (b) the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).

7. The fusion protein of claim 1 , wherein the fusion protein comprises mCD4-hMyD88 as set forth in SEQ ID NO:21 or hCD4-hMyD88 as set forth in SEQ ID NO:20.

8. The fusion protein of claim 1 , wherein the TCR is the DMFS TCR having the amino acid sequence of residues 1-603 of SEQ ID NO:22 and the region of MyD88 lacking the TIR domain corresponds to amino acids 1-155 of human MyD88 (SEQ ID NO:24).

9. The fusion protein of claim 1 , wherein the fusion protein comprises hTCR-hMyD88 as set forth in SEQ ID NO:22.

10. An isolated population of cells expressing at least one fusion protein of claim 1 .

11. The isolated population of cells of claim 10 , wherein the isolated population of cells expresses at least one fusion protein selected from the group consisting of mCD8α-hMyD88 (SEQ ID NO:17), hCD8α-hMyD88 (SEQ ID NO:14), hCD8αTM-hMyD88 (SEQ ID NO:18), mCD4-hMyD88 (SEQ ID NO:21), hCD4-hMyD88 (SEQ ID NO:20) and hTCR-hMyD88 (SEQ ID NO:22).

12. A method of treating a subject having cancer or an infectious disease, comprising administering to a subject having cancer or an infectious disease a therapeutically-effective amount of at least one population of cells as defined in claim 10 .

13. A method of treating a subject having cancer or an infectious disease, comprising administering to a subject having cancer or an infectious disease a therapeutically-effective amount of at least one population of cells as defined in claim 11 .

14. A method of conferring T cell resistance against myeloid derived suppressor cells (MDSC)-mediated suppression, comprising expressing at least one fusion protein of claim 1 in a T cell.

15. The method of claim 14 , wherein the fusion protein is selected from the group consisting of mCD8α-hMyD88 (SEQ ID NO:17), hCD8α-hMyD88 (SEQ ID NO:14), hCD8αTM-hMyD88 (SEQ ID NO:18), mCD4-hMyD88 (SEQ ID NO:21), hCD4-hMyD88 (SEQ ID NO:20) and hTCR-hMyD88 (SEQ ID NO:22).

16. A method of enhancing immune cell recognition of an antigen, comprising expressing at least one fusion protein of claim 1 in an immune cell.

17. The method of claim 16 , wherein the fusion protein is selected from the group consisting of mCD8α-hMyD88 (SEQ ID NO:17), hCD8α-hMyD88 (SEQ ID NO:14), hCD8αTM-hMyD88 (SEQ ID NO:18), mCD4-hMyD88 (SEQ ID NO:21), hCD4-hMyD88 (SEQ ID NO:20) and hTCR-hMyD88 (SEQ ID NO:22).

18. The method of claim 16 , wherein the antigen is present at a low concentration in vitro or in vivo.

19. The method of claim 16 , wherein the antigen is a weakly antigenic antigen.

Assignees

• Davila Eduardo
• Univ Maryland

Inventors

• Davila Eduardo

Classifications

• A61K40/4273

  Glycoprotein 100 [Gp100] · CPC title

• A61K40/4272

  Melan-A/MART · 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

• A61K2239/38

  characterised by the dose, timing or administration schedule · CPC title