# US 2013/0280269 A1 · Manipulation of HSP70 and IRE1Alpha Protein Interactions

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- Number: US 2013/0280269 A1
- Document ID: US20130280269A1
- Status: Published application
- Kind: A1
- Application number: 13/702,649
- Filed: 2011-06-08
- Published: 2013-10-24
- Earliest claimed: 2010-06-08
- Assignees: National University of Ireland
- Inventors: Afshin Samali; Sanjeev Gupta; Aysawaria Deepti
- Web page: https://patentgrep.com/patent/US20130280269A1

## CPC

- C12N 9/12: transferring phosphorus containing groups, e.g. kinases (2.7)
- C07K 14/47: from mammals
- C12N 5/0018: {Culture media for cell or tissue culture (media for specific animal cell type C12N5/06)}
- C12N 2501/07: Heat shock proteins
- C12N 2510/02: Cells for production
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)

## Abstract

The present invention relates to methods that manipulate the level of interaction between the proteins Hsp70 and IRE1α. In particular, the invention relates to methods of manipulating this interaction in order to increase protein yield, and to methods of manipulating this interaction in order to treat diseases associated with abnormal apoptotic activity, including cancer and autoimmune diseases.

## Claims

### Claim at position 1

1-75. (canceled)

### Claim 76

A method of generating a cell for protein production, the method comprising manipulating the cell to alter a level of interaction between Hsp70 and IRE1α.

### Claim 77 (depends on 76)

The method of claim 76, wherein the method comprises manipulating the cell to increase the level of interaction between Hsp70 and IRE1α, such that the level of unconventionally-spliced XBP1 (XBP1s) mRNA rises.

### Claim 78 (depends on 77)

The method of claim 77, wherein the method comprises increasing a level of expression of at least one of Hsp70 or IRE1α.

### Claim 79 (depends on 78)

The method of claim 78, wherein the method comprises transfecting the cell with a plasmid expressing Hsp70 or IRE1α.

### Claim 80 (depends on 78)

The method of claim 78, wherein the method comprises inserting a strong promoter that is transcriptionally linked to an endogenous gene encoding Hsp70 or IRE1α.

### Claim 81 (depends on 76)

The method of claim 76, wherein the method comprises manipulating the cell to decrease the level of interaction between Hsp70 and IRE1α, such that level of activated JNK or level of activation of JNK decreases.

### Claim 82 (depends on 76)

The method of claim 76, wherein the method comprises administering at least one of an Hsp70 neutralizing agent or an IRE1α neutralizing agent to the cell.

### Claim 83 (depends on 76)

The method of claim 76, wherein the Hsp70 is Hsp72.

### Claim 84 (depends on 76)

The method of claim 76, further comprising isolating a protein of interest from a preparation of the cell.

### Claim 85 (depends on 76)

A cell produced according to the method of claim 76.

### Claim 86

A method of treating a disease, the method comprising modulating an interaction between Hsp70 and IRE1α.

### Claim 87 (depends on 86)

The method of claim 86, wherein the method comprises modulating the interaction between Hsp70 and IRE1α such that a level of unconventionally-spliced XBP1 (XBP1s) mRNA is altered.

### Claim 88 (depends on 87)

The method of claim 87, wherein the method comprises:
decreasing a level of the interaction between Hsp70 and IRE1α, such that the level of the unconventionally-spliced XBP1 (XBP1s) mRNA is reduced; or
increasing a level of the interaction between Hsp70 and IRE1α, such that the level of the unconventionally-spliced XBP1 mRNA rises.

### Claim 89 (depends on 88)

The method of claim 88, wherein the disease is selected from the group consisting of cancer, an autoimmune disease, diabetes and cardiac disease.

### Claim 90 (depends on 88)

The method of claim 88, comprising administering at least one of an Hsp70 neutralizing agent or an IRE1α neutralizing agent to an affected cell.

### Claim 91 (depends on 88)

The method of claim 88, wherein the Hsp70 is Hsp72.

### Claim 92 (depends on 90)

The method of claim 90, wherein the Hsp70 neutralizing agent is an anti-Hsp70 antibody, and wherein the IRE1α neutralizing agent is an anti-IRE1α antibody.

### Claim 93 (depends on 92)

The method of claim 92, wherein the anti-Hsp70 antibody is an anti-HSP70 monoclonal antibody and wherein the anti-IRE1αantibody is an anti-IRE1α monoclonal antibody.

### Claim 94 (depends on 92)

The method of claim 92, wherein the anti-Hsp70 antibody is directed to an ATPase domain of Hsp70.

### Claim 95 (depends on 92)

The method of claim 92, wherein the anti-IRE1α antibody is directed to a cytosolic C-terminal region of IRE1α.

### Claim 96 (depends on 88)

The method of claim 88, wherein the method further comprises inducing cellular stress.

### Claim 97 (depends on 86)

The method of claim 86, wherein the method comprises modulating the interaction between Hsp70 and IRE1α such that a level of activated JNK or a level of JNK activation is altered.

### Claim 98

A pharmaceutical composition comprising an Hsp70 neutralizing agent and/or an IRE1α neutralizing agent and a pharmaceutically acceptable carrier or the amino acid sequence of Hsp70 or a plasmid encoding Hsp70, and/or the amino acid sequence of IRE1α or a plasmid encoding IRE1α and a pharmaceutically acceptable carrier.

### Claim 99 (depends on 98)

A method of treating a disease comprising administering the pharmaceutical composition of claim 98 to an animal, wherein the disease selected from the group consisting of cancer, an autoimmune disease, diabetes and cardiac disease.