Markers and Methods for Assessing and Treating Ulcerative Colitis and Related Disorders Using 66 Gene Panel

• US 20090054253A1
• Filed: 08/30/2007
• Published: 02/26/2009
• Est. Priority Date: 08/30/2006
• Status: Active Grant

• Current Assignee: Centocor Ortho Biotech Incorporated (Johnson & Johnson)
• Original Assignee: Centocor Ortho Biotech Incorporated (Johnson & Johnson)
• Inventors: Song, Xiao-yu, Li, Xilin
• Granted Patent: US 7,943,310 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• 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 for prognostic or diagnostic assessment of a gastrointestinal-related disorder, such as ulcerative colitis, in a subject correlates the presence, absence, and/or magnitude of a gene in a sample with a reference standard to determine the presence and/or severity of the disorder, and/or the response to treatment for the disorder. The method enables identification of the effectiveness of candidate therapies.

Claims

1. A method for prognostic or diagnostic assessment of a gastrointestinal-related disorder in a subject, comprising:

a) preparing a sample of nucleic acids from a specimen obtained from the subject;

b) contacting the sample with a panel of nucleic acid segments consisting of at least 2 genes represented by nucleic acids from the group consisting of SEQ ID NOS;

1-66 to detect the levels of the panel segments;

c) evaluating the sample against a reference standard to determine the magnitude of change in the amounts of the at least 2 members present in the sample; and

d) correlating the magnitude of change with the presence or resolution of the gastrointestinal-related disorder.

2. The method of claim 1, wherein the subject is a patient having a gastrointestinal-related disorder and steps a) through d) are performed before, during, and/or after treatment of the patient with a therapy for the gastrointestinal-related disorder.

3. The method of claim 2, wherein steps a) through d) are performed during treatment of the patient with a therapy for the gastrointestinal-related disorder and about weeks after commencement of treatment.

4. The method of claim 2, wherein the gastrointestinal-related disorder is ulcerative colitis.

5. The method of claim 2, wherein the reference standard is from the group consisting of colon biopsy from a normal patient, colon biopsy from an untreated ulcerative colitis patient, and colon biopsy from a treated ulcerative colitis patient.

6. The method of claim 2, wherein the reference standard is from the subject prior to treatment with a therapy, the sample of nucleic acids is from the subject after treatment with a therapy, and the correlating step evaluates the effectiveness of treatment with the therapy.

7. The method of claim 2, wherein the therapy is an anti-TNFα

antibody,

8. The method of claim 1, wherein the collection is an array of nucleic acid segments.

9. The method of claim 2, wherein the sample is from a colon biopsy of a patient selected from the group consisting of patients suspected of having ulcerative colitis, patients diagnosed with ulcerative colitis undergoing treatment with an approved agent, and patients diagnosed with ulcerative colitis undergoing treatment with an experimental agent.

10. The method of claim 2, wherein the sample is from a source selected from the group consisting of a patient providing the sample prior to administration of a therapy, a placebo treated patient having a gastrointestinal-related disorder, and a sample from a biobank.

11. The method of claim 1, wherein the at least one gene from the collection is selected from the group consisting of cytokines, chemokines, transcription factors, proteases, protease inhibitors, structural and adhesion molecules, and genes for proteins involved in lipid metabolism.

12. The method of claim 1, wherein the sample comprises a colon biopsy sample,

13. The method of claim 1, wherein the sample comprises peripheral blood cells.

14. The method of claim 1, wherein the sample is contacted with a panel of nucleic acid segments comprising at least 4 members from the group consisting of SEQ ID NOS:

1-66.

15. The method of claim 14, wherein the at least four nucleic acid segments are representative of or selected from an innate or adaptive immune response-related gene selected from the group consisting of SEQ ID NOS:

1-26, 29, 30, and 38;

a cell-cell interaction, cell-matrix interaction or matrix regulation-related gene selected from the group consisting of SEQ ID NOS;

27, 34, 37,49, and 51;

a cell-cell, intracellular signaling pathway-related gene selected from the group consisting of SEQ ID NOS;

28, 39, and 41-48;

a cell growth and apoptosis-related gene selected from the group consisting of SEQ ID NOS;

31-33, 35, and 36;

a protein regulation-related gene selected from the group consisting of SEQ ID NOS. 50 and 66;

a metabolic regulation-related gene selected from the group consisting of SEQ ID NOS;

52-61, 64, and 65; and

a cytoskeleton organization-related gene selected from the group consisting of SEQ ID NOS;

62 and 63.

16. The method of claim 1, wherein at least one of the at least two nucleic acid segments is representative of or selected from the group consisting of SEQ ID NOS:

1-26, 29, 30, and 38.

17. The method of claim 1, wherein the at least two gene segments are representative of or selected from the group consisting of SEQ ID NOS:

1-26, 29, 30, and 38 and SEQ ID NOS;

31-33, 35, and 36.

48. The method of claim 1, wherein the subject is a patient having a gastrointestinal-related disorder and steps a) through d) are performed before, during, and/or after treatment of the patient with a therapy for the gastrointestinal-related disorder.

18. A method for prognostic or diagnostic assessment of a gastrointestinal-related disorder in a subject, comprising:

a) preparing a sample of nucleic acids from a sample obtained from a patient;

b) contacting the sample with a panel of nucleic acid segments represented by nucleic acids consisting of at least one member from the group consisting of SEQ ID NOS;

1-26,29, 30, and 38 to defect the presence of the panel segments;

c) evaluating the sample against a reference standard to determine the change and/or magnitude of change in the expression level of the amounts of the at least one member present in the sample; and

d) correlating the change and/or magnitude of expression level with the presence or resolution of the gastrointestinal-related disorder.

19. An array-based testing method for prognostic or diagnostic assessment of a gastrointestinal-related disorder in a patient, comprising:

a) preparing a mixture of nucleic acids from a specimen obtained from a patient;

b) labeling said specimen nucleic acids with a detectable marker to form a sample;

c) contacting the sample with an array comprising a plurality of nucleic acid segments, wherein each nucleic acid segment is immobilized to a discrete and known address on a substrate surface of the array, wherein at least two members of a gastrointestinal-related gene panel consisting of SEQ ID NOS;

1-66 are identified as features of the array by address, and wherein said array further comprises at least one calibration nucleic acid at a known address on the substrate;

d) determining the degree of binding of the specimen nucleic acids to the nucleic acid segments; and

e) comparing the degree of binding to a reference standard to enable a prognostic or diagnostic assessment,

20. The method of claim 19, further comprising the step of performing a statistical comparison of the specimen nucleic acids from gastrointestinal-related disorder patients treated with a therapy to a reference standard to evaluate the effect of treatment with the therapy,

21. The method of claim 20, wherein the gastrointestinal-related disorder is ulcerative colitis and the gastrointestinal-related gene panel is an ulcerative colitis-related gene panel.

22. The method of claim 20, wherein the therapy is an anti-TNFα

antibody.

23. The method of claim 19, wherein the specimen is from a colon biopsy of a patient selected from the group consisting of patients suspected of having ulcerative colitis, patients diagnosed with ulcerative colitis not undergoing treatment, and patients diagnosed with ulcerative colitis undergoing treatment with a therapy.

24. The method of claim 19, wherein the specimen is from a source selected from the group consisting of a patient providing the specimen prior to administration of a therapy, a patient having a similar disease or condition treated with a placebo, and a sample from a biobank.

25. The method of claim 19, wherein the members of the gene panel are selected from the group consisting of cytokines, chemokines, transcription factors, proteases, protease inhibitors, structural and adhesion molecules, and genes for proteins involved in lipid metabolism.

26. The method of claim 19, wherein the specimen comprises a colon biopsy sample.

27. The method of claim 19, wherein the specimen comprises peripheral blood cells.

28. The method of claim 21, wherein the comparing the degree of binding step further comprises a stringent test of the similarity of feature intensity changes of the array of the ulcerative colitis-related gene panel.

29. A reagent for testing the responsiveness of a cell or subject to a therapy for a gastrointestinal-related disorder, comprising at least one member selected from the group consisting of an oligonucleotide comprising at least 15 nucleotides complementary to a nucleotide sequence of one of SEQ ID NOS:

1-66, a polypeptide encoded by at least a portion of one of SEQ ID NOS;

1-66, and a ligand for the polypeptide encoded by at least a portion of one of SEQ ID NOS;

1-66.

30. The reagent of claim 29, wherein the gastrointestinal-related disorder is ulcerative colitis.

31. A method of testing for responsiveness to a therapy for a gastrointestinal-related disorder in a patient sample comprising contacting the patient sample with the reagent of claim 29 and comparing the levels of at least a portion of one of the genes or proteins of SEQ ID NOS:

1-66 to a reference standard.

32. The method of claim 31, wherein the testing is done by RT-PCR,

33. The method of claim 31, wherein the testing is done by ELISA.

34. A method of testing the effectiveness of a therapy for a gastrointestinal-related disorder, comprising:

a) contacting a sample from a patient being treated for the gastrointestinal-related disorder with the reagent of claim 29;

b) measuring levels of the at least one member; and

c) correlating the levels of the at least one member with the effectiveness of the therapy,

35. The method of claim 34, wherein the correlating step comprises comparing the levels with levels of the at least one member of a sample from the patient prior to treatment with the therapy and wherein a decrease of at least about 2-fold in the level of the at least one member from the patient being treated versus the patient prior to treatment indicates a responder to the therapy.

36. The method of claim 35, wherein the gastrointestinal-related disorder is ulcerative colitis.

37. The method of claim 35, wherein the therapy comprises an antagonist of TNFα

.

38. The method of claim 37, wherein the antagonist is an antibody to TNFα

.

39. The method of claim 38, wherein the antibody to TNFα

is infliximab.

42. A method of testing the effectiveness of a therapy for ulcerative colitis, comprising:

a) contacting a sample from a patient being treated for ulcerative colitis with at least two members of the reagent of claim 29;

b) measuring levels of the at least two members; and

c) correlating the levels of the at least two members with the effectiveness of the therapy.

43. The method of claim 42, wherein the correlating step comprises comparing the levels with levels of the at least two members of a sample from the patient prior to treatment with the therapy and wherein a decrease of at least about 2-fold in the level of the at least two members from the patient being treated versus the patient prior to treatment indicates a responder to the therapy.

44. The method of claim 43, wherein the therapy comprises an antagonist of TNFα

.

45. The method of claim 44, wherein the antagonist is an antibody to TNFα

.

46. The method of claim 45, wherein the antibody to TNFα

is infliximab.

40. A kit for prognostic or diagnostic use, comprising an oligonucleotide comprising at least 15 nucleotides complementary to a polynucleotide comprising the nucleotide sequence of a marker gene or the complementary strand thereof and cells expressing the master gene, wherein the marker gene is selected from the group consisting of SEQ ID NOS:

1-66.

41. A kit for screening for a therapeutic agent for UC, the kit comprising an antibody which recognizes a peptide comprising an amino acid sequence encoded by a marker gene and ceils expressing the marker gene, wherein the marker gene is selected from the group consisting of SEQ ID NOS:

1-66.

47. A method for prognostic or diagnostic assessment of a gastrointestinal-related disorder in a subject, comprising:

d) preparing a sample of nucleic acids from a specimen obtained from the subject;

e) contacting the sample with a panel of nucleic acid segments consisting of at least 2 members from the group of genes represented by nucleic acids selected from the group consisting of SEQ ID NOS;

1-66 to detect the levels of the panel segments;

f) evaluating the sample against a reference standard to determine the magnitude of change in the amounts of the at least 2 members present in the sample; and

g) correlating the magnitude of change with the presence or resolution of the gastrointestinal-related disorder.

49. Any invention described herein.