Methods and compositions relating to modulation of hepatocyte growth, plasma cell differentiation or T cell subset activity by modulation of XBP-1 activity

• US 6,632,608 B2
• Filed: 12/29/2000
• Issued: 10/14/2003
• Est. Priority Date: 12/30/1999
• Status: Expired due to Term

• Current Assignee: Cornell University
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Reimold, Andreas M., Glimcher, Laurie H.
• Primary Examiner(s): Wehbé , Anne M.
• Assistant Examiner(s): LI, QIAN JANICE
• Application Number:
  US09/753,346
• Publication Number:
  US 20020059652A1
• Time in Patent Office: 1,019 Days
• Field of Search: 435/69.1, 435/4, 435/6, 435/7.1, 435/372, 435/372.2, 435/372.3, 536/23.5, 536/24.1
• US Class Current: 506/10
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 2267/0306 Animal model for genetic diseases
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C07K 14/4753 Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: G01N 2333/4753 Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 involving specific cell types
• CPC Class Codes: G01N 33/505 involving T-cells
• CPC Class Codes: G01N 33/5067 Liver cells
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/6845 : Methods of identifying protein-protein interactions in protein mixtures
• CPC Class Codes: G01N 33/6863 : Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• CPC Class Codes: G01N 33/74 : involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: Y10S 435/973 : Simultaneous determination of more than one analyte

Abstract

The invention demonstrates that the transcription factor XBP-1 is a regulator of hepatocyte growth, plasma cell differentiation and T cell subset activity. Methods for identifying modulators of hepatocyte growth, plasma cell differentiation and/or T cell subset activity, using XBP-1-containing indicator compositions or XBP-1-deficient cells, are provided. Methods of modulating hepatocyte growth, plasma cell differentiation and/or T cell subset activity (e.g., Th2 cytokine production) using agents that modulate the activity of XBP-1 are also provided. Methods for diagnosing disorders associated with aberrant hepatocyte growth, plasma cell differentiation and/or T cell subset activity, by assessing a change in XBP-1 expression, are also provided. XBP-1 deficient cells, animals and embryos, as well as kits for the methods of the invention, are also provided by the invention.

Claims

1. A method of identifying a compound that modulates hepatocyte cell growth comprising

a) providing an indicator composition comprising an X-box-binding protein (XBP-1);

b) contacting the indicator composition with each member of a library of test compounds;

c) selecting from the library of test compounds a compound of interest that modulates the activity of the XBP-1 protein; and

d) determining the effect of the compound of interest on hepatocyte growth to thereby identify a compound that modulates hepatocyte cell growth.

2. The method of claim 1, wherein the indicator composition is a cell that expresses the XBP-1 protein.

3. The method of claim 2, wherein the cell has been engineered to express the XBP-1 protein by introducing into the cell an expression vector encoding the XBP-1 protein.

4. The method of claim 1, wherein the indicator composition is a cell free composition.

5. The method of claim 1, wherein the indicator composition is a cell that expresses an XBP-1 protein and a target molecule, and the ability of the test compound to modulate the interaction of the XBP-1 protein with a target molecule is monitored.

6. The method of claim 1, wherein the indicator composition comprises an indicator cell, wherein the indicator cell comprises an XBP-1 protein and a reporter gene, wherein the reporter gene is operably linked to an XBP-1 responsive element.

7. The method of claim 6, wherein said indicator cell contains:

a recombinant expression vector encoding the XBP-1 protein; and

a vector comprising an XBP-1 responsive regulatory element operatively linked to a reporter gene; and

said method comprises;

a) contacting the indicator cell with a test compound;

b) determining the level of expression of the reporter gene in the indicator cell in the presence of the test compound; and

c) comparing the level of expression of the reporter gene in the indicator cell in the presence of the test compound with the level of expression of the reporter gene in the indicator cell in the absence of the test compound whereby a difference in the level of the reporter gene expression indicates that the compound modulates XBP-1 protein.

8. The method of claim 6, wherein the XBP-1 responsive element is selected from the group consisting of:

α

-1 antitrypsin, α

-fetoprotein, HLA DRα

, and the 21 base pair repeat enhancer of HTLV-1LTR.

9. A method of identifying a compound that modulates plasma cell differentiation comprising

a) providing an indicator composition comprising an X-box-binding protein (XBP-1) protein;

b) contacting the indicator composition with each member of a library of test compounds;

c) selecting from the library of test compounds a compound of interest that modulates the activity of the XBP-1 protein; and

d) determining the effect of the compound of interest on plasma cell differentiation to thereby identify a compound that modulates plasma cell differentiation.

10. The method of claim 9, wherein the indicator composition is a cell that expresses the XBP-1 protein.

11. The method of claim 10, wherein the cell has been engineered to express the XBP-1 protein by introducing into the cell an expression vector encoding the XBP-1 protein.

12. The method of claim 9, wherein the indicator composition is a cell free composition.

13. The method of claim 9, wherein the indicator composition is a cell that expresses an XBP-1 protein and a target molecule, and the ability of the test compound to modulate the interaction of the XBP-1 protein with a target molecule is monitored.

14. The method of claim 9, wherein the indicator composition comprises an indicator cell, wherein the indicator cell comprises an XBP-1 protein and a reporter gene, wherein the reporter gene is operably linked to an XBP-1 responsive element.

15. The method of claim 14, wherein said indicator cell contains;

a recombinant expression vector encoding the XBP-1 protein; and

a vector comprising an XBP-1 responsive regulatory element operatively linked to a reporter gene; and

said method comprises;

a) contacting the indicator cell with a test compound;

b) determining the level of expression of the reporter gene in the indicator cell in the presence of the test compound; and

c) comparing the level of expression of the reporter gene in the indicator cell in the presence of the test compound with the level of expression of the reporter gene in the indicator cell in the absence of the test compound whereby a difference in the level of the reporter gene expression indicates that the compound modulates XBP-1 protein.

16. The method of claim 14, wherein the XBP-1 responsive element is selected from the group consisting of:

α

-1 antitrypsin, α

-fetoprotein, HLA DRα

, and the 21 base pair repeat enhancer of HTLV-1LTR.

17. A method of identifying a compound that modulates TH2 cell subset activity comprising:

a) providing an indicator composition comprising an X-box-binding protein (XBP-1);

b) contacting the indicator composition with each member of a library of test compounds;

c) selecting from the library of test compounds a compound of interest that modulates the activity of the XBP-1 protein; and

d) determining the effect of the compound of interest on T cell subset activity to thereby identify a compound that modulates TH2 cell subset activity.

18. The method of claim 17, wherein the indicator composition is a cell that expresses the XBP-1 protein.

19. The method of claim 18, wherein the cell has been engineered to express the XBP-1 protein by introducing into the cell an expression vector encoding the XBP-1 protein.

20. The method of claim 17, wherein the indicator composition is a cell free composition.

21. The method of claim 17, wherein the indicator composition is a cell that expresses an XBP-1 protein and a target molecule, and the ability of the test compound to modulate the interaction of the XBP-1 protein with a target molecule is monitored.

22. The method of claim 17, wherein the indicator composition comprises an indicator cell, wherein the indicator cell comprises an XBP-1 protein and a reporter gene, wherein the reporter gene is operably linked to an XBP-1 responsive element.

23. The method of claim 22, wherein said indicator cell contains;

a recombinant expression vector encoding the XBP-1 protein; and

a vector comprising an XBP-1 responsive regulatory element operatively linked to a reporter gene; and

said method comprises;

a) contacting the indicator cell with a test compound;

b) determining the level of expression of the reporter gene in the indicator cell in the presence of the test compound; and

c) comparing the level of expression of the reporter gene in the indicator cell in the presence of the test compound with the level of expression of the reporter gene in the indicator cell in the absence of the test compound whereby a difference in the level of the reporter gene expression indicates that the compound modulates XBP-1 protein.

24. The method of claim 22, wherein the XBP-1 responsive element is selected from the group consisting of:

α

-1 antitrypsin, α

-fetoprotein, HLA DRα

, and the 21 base pair repeat enhancer of HTLV-1LTR.

25. The method of claim 22, wherein T cell cytokine production is measured.

26. The method of claim 17, wherein T cell cytokine production is measured.