Methods regarding enhanced T-cell receptor-mediated tumor necrosis factor superfamily mRNA expression in peripheral blood leukocytes in patients with crohn's disease

• US 7,838,239 B2
• Filed: 04/05/2007
• Issued: 11/23/2010
• Est. Priority Date: 04/07/2006
• Status: Expired due to Fees

• Current Assignee: Cedars-Sinai Medical Center (Cedars Sinai Health System)
• Original Assignee: Cedars-Sinai Medical Center (Cedars Sinai Health System), Hitachi Chemical Company Limited (Resonac Holdings Corp.), Hitachi Chemical Research Center, Inc (Resonac Holdings Corp.)
• Inventors: Mitsuhashi, Masato, Targan, Stephan R.
• Primary Examiner(s): Landsman; Robert
• Application Number:
  US12/296,425
• Publication Number:
  US 20090253133A1
• Time in Patent Office: 1,328 Days
• Field of Search: None
• US Class Current: 435/6.16
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/106 Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

A method is disclosed for determining whether a human having Crohn'"'"'s disease is likely to respond to a therapy targeting a TNFSF member or a cytokine by measuring the level of certain mRNAs in response to a stimulus. A method of evaluating the effectiveness of a Crohn'"'"'s disease therapy in a human is also disclosed. Furthermore, a method of screening compounds for use in the treatment of Crohn'"'"'s disease is disclosed. A method of monitoring the disease state over time in Crohn'"'"'s disease patients is also disclosed.

Claims

1. A method of determining whether a human having Crohn'"'"'s disease is likely to respond to immunosuppressive therapy, comprising:

stimulating leukocytes with an anti-T-cell receptor antibody in vitro in a first sample that comprises leukocytes from the human;

after the stimulation, measuring the amount of an mRNA encoding a tumor necrosis factor superfamily member in the first sample;

exposing leukocytes in vitro in a second sample comprising leukocytes from the human to a control stimulus;

measuring the amount of the mRNA in the second sample after exposing; and

determining a ratio of the amount of the mRNA in the first sample to the amount of the mRNA in the second sample, wherein a ratio of at least about 1.7;

1 indicates that the human is likely to respond to said immunosuppressive therapy.

2. The method of claim 1, wherein at least one of the first and second samples comprises whole blood.

3. The method of claim 1, wherein the control stimulus comprises a purified control immunoglobulin.

4. The method of claim 1, wherein the therapy targets TNF-α

activity.

5. The method of claim 4, wherein the therapy comprises administration of infliximab.

6. The method of claim 1, wherein the therapy comprises administration of an agent selected from the group consisting of cyclosporine A and tacrolimus.

12. The method of any of claims 1 and 7 wherein the tumor necrosis factor superfamily (“

TNFSF”

) member is selected from the group consisting of TNFSF-2, TNFSF-5, TNFSF-6, and TNFSF-14.

13. The method of claim 12, wherein the TNFSF member is selected from the group consisting of TNFSF-2 and TNFSF-14.

7. A method of evaluating the state of Crohn'"'"'s disease in a human, comprising:

stimulating leukocytes with an anti-T-cell receptor antibody in vitro in a first sample that comprises leukocytes and is obtained at a first time from the human;

after the stimulation, measuring the amount of an mRNA encoding a tumor necrosis factor superfamily member in the first sample;

exposing leukocytes in vitro to a control stimulus in a second sample comprising leukocytes obtained from the human at the first time;

measuring the amount of the mRNA in the second sample after exposing;

determining a first ratio of the amount of the mRNA in the first sample to the amount of the mRNA in the second sample;

stimulating leukocytes with an anti-T-cell receptor antibody in vitro in a third sample that comprises leukocytes and is obtained from the human at a second time that is subsequent to the first time;

after the stimulation, measuring the amount of the mRNA in the third sample;

exposing leukocytes in vitro to a control stimulus in a fourth sample comprising leukocytes obtained from the human at the second time;

measuring the amount of the mRNA in the fourth sample after exposing;

determining a second ratio of the amount of the mRNA in the third sample to the amount of the mRNA in the fourth sample; and

comparing the first and second ratios, wherein a statistically significant difference in the first and second ratios is indicative of a change in the disease state.

8. The method of claim 7, wherein the control stimulus comprises a purified control immunoglobulin.

9. The method of claim 7, wherein at least one of the first, second, third and fourth samples comprises whole blood.

10. The method of claim 7, wherein the significant difference in the ratios is that the second ratio is greater than the first ratio, and the change in disease state is a progression of the disease.

11. The method of claim 7, wherein the significant difference in the ratios is that the first ratio is greater than the second ratio, and the change in disease state is a regression of the disease.