التفاصيل البيبلوغرافية
| العنوان: |
Methods of testing for bronchial asthma or chronic obstructive pulmonary disease |
| Document Number: |
20050208496 |
| تاريخ النشر: |
September 22, 2005 |
| Appl. No: |
10/631467 |
| Application Filed: |
July 31, 2003 |
| مستخلص: |
An objective of the present invention is to provide a method of testing for bronchial asthma or chronic obstructive pulmonary disease, a method of screening for candidate compounds for treating bronchial asthma or chronic obstructive pulmonary disease, and a pharmaceutical agent for treating bronchial asthma or chronic obstructive pulmonary disease. The present invention identified genes whose expression levels varied between respiratory epithelial cells that had been stimulated by IL-13 to induce the goblet cell differentiation, and unstimulated respiratory epithelial cells. The respiratory epithelial cells were cultured according to the air interface method. The genes were revealed to be useful as markers for testing for bronchial asthma or chronic obstructive pulmonary disease and screening for therapeutic agents for such diseases. Specifically, the present invention provides methods of testing for bronchial asthma or chronic obstructive pulmonary disease and methods of screening for compounds to treat the diseases based on the comparison of the expression levels of marker genes identified as described above. |
| Inventors: |
Ohtani, Noriko (Gunma, JP); Sugita, Yuji (Tsukuba-shi, JP); Yamaya, Mutsuo (Sendai-shi, JP); Kubo, Hiroshi (Sendai-shi, JP); Nagai, Hiroichi (Gifu-shi, JP); Izuhara, Kenji (Saga-shi, JP) |
| Assignees: |
Genox Research, Inc. (Ibaraki, JP) |
| Claim: |
1. A method of testing for bronchial asthma or chronic obstructive pulmonary disease, which comprises the steps of: (1) determining the expression level of a marker gene in a biological sample from a subject; (2) comparing the expression level determined in step (1) with the expression level of the marker gene in a biological sample from a healthy subject; and (3) judging the subject to have bronchial asthma or chronic obstructive pulmonary disease when the result of the comparison in step (2) indicates that (i) the expression level of the marker gene in the subject is higher than that in the control when the marker gene is a gene according to (a) or (ii) when the expression level of the marker gene in the subject is lower than that in the control when said marker gene is a gene according to (b); wherein the marker gene is any one selected from the group according to (a) or (b): (a) a group of genes whose expression levels increase when respiratory epithelial cells are stimulated with interleukin-13, and comprise any one of the nucleotide sequences of SEQ ID NOs: 25 to 310; (b) a group of genes whose expression levels decrease when respiratory epithelial cells are stimulated with interleukin-13, and comprise any one of the nucleotide sequences of SEQ ID NOs: 311 to 547. |
| Claim: |
2. The testing method according to claim 1, wherein the biological sample is a respiratory epithelial cell. |
| Claim: |
3. The testing method according to claim 1, wherein the gene expression level is measured by PCR analysis of the cDNA. |
| Claim: |
4. The testing method according to claim 1, wherein the gene expression level is measured by detecting the protein encoded by the marker gene. |
| Claim: |
5. A reagent for testing for bronchial asthma or chronic obstructive pulmonary disease, wherein the reagent comprises a polynucleotide comprising the nucleotide sequence of a marker gene, or an oligonucleotide having at least 15 nucleotides and comprising a nucleotide sequence complementary to the complementary strand of the nucleotide sequence of the marker gene, and wherein, the marker gene is any one selected from the group according to (a) or (b) in claim 1. |
| Claim: |
6. A reagent for testing for bronchial asthma or chronic obstructive pulmonary disease, wherein the reagent comprises an antibody that recognizes a protein encoded by a marker gene, and wherein the marker gene is any one selected from the group according to (a) or (b) in claim 1. |
| Claim: |
7. A method of screening for a therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, wherein the marker gene is any one selected from the group according to (a) or (b) in claim 1, and wherein the method comprises the steps of: (1) contacting a candidate compound with a cell expressing the marker gene; (2) measuring the expression level of said gene; and (3) selecting a compound that decreases the expression level of a marker gene belonging to group (a) or increases the expression level of a marker gene belonging to group (b), as compared to that in a control with which the compound has not been contacted. |
| Claim: |
8. The method according to claim 7, wherein the cell is a respiratory epithelial cell or a goblet cell. |
| Claim: |
9. The method according to claim 8, which comprises the step of culturing the respiratory epithelial cells under the condition in which culture medium is removed from the apical side of said cells and the culture medium is supplied from the basolateral side of the cells. |
| Claim: |
10. A kit for screening for a candidate compound for a therapeutic agent to treat bronchial asthma or chronic obstructive pulmonary disease, wherein the kit comprises (i) a polynucleotide comprising the nucleotide sequence of a marker gene, or an oligonucleotide having at least 15 nucleotides and comprising a nucleotide sequence that is complementary to the complementary strand of the polynucleotide, and (ii) a cell expressing the marker gene, and wherein the marker gene is any one selected from the group according to (a) or (b) in claim 1. |
| Claim: |
11. A kit for screening for a candidate compound for a therapeutic agent to treat bronchial asthma or chronic obstructive pulmonary disease, wherein the kit comprises (i) an antibody that recognize a protein encoded by a marker gene, and (ii) a cell expressing the marker gene, wherein the marker gene is selected from the group according to (a) or (b) in claim 1. |
| Claim: |
12. The kit according to claim 10, which further comprises a cell-supporting material to culture respiratory epithelial cells under conditions in which the culture medium is supplied from the basolateral side of the cells. |
| Claim: |
13. The kit according to claim 12, which further comprises respiratory epithelial cells. |
| Claim: |
14. An animal model for bronchial asthma or chronic obstructive pulmonary disease, wherein the animal is a transgenic nonhuman vertebrate wherein the expression level of a marker gene, or a gene functionally equivalent to the marker gene, has been increased in the respiratory tissue, wherein the marker gene is any one selected from the group according to (a) in claim 1 or the following (A): (A) a group of genes whose expression levels increase in the lung of an animal model for bronchial hypersensitivity induced by an exposure to the ovalbumin antigen, wherein the genes comprise any one of the nucleotide sequences of SEQ ID NOs: 954 to 1174. |
| Claim: |
15. The animal model according to claim 14, wherein the nonhuman vertebrate is a mouse. |
| Claim: |
16. An animal model for bronchial asthma or chronic obstructive pulmonary disease, wherein the animal is a transgenic nonhuman vertebrate wherein the expression level of a marker gene, or a gene functionally equivalent to the marker gene, has been decreased in the respiratory tissue, wherein the marker gene is any one selected from the group according to (b) in claim 1 or the following (B): (B) a group of genes whose expression levels decrease in the lung of an animal model for bronchial hypersensitivity induced by an exposure to the ovalbumin antigen, wherein the genes comprise any one of the nucleotide sequences of SEQ ID NOs: 1376 to 1515. |
| Claim: |
17. The animal model according to claim 16, wherein the nonhuman vertebrate is a mouse. |
| Claim: |
18. A method for producing an animal model for bronchial asthma or chronic obstructive pulmonary disease, which comprises the step of administering to a mouse any one of (i) or (ii): (i) a polynucleotide comprising the nucleotide sequence constituting any one of the genes selected from the gene group according to (A) in claim 14; and (ii) a protein encoded by a polynucleotide comprising the nucleotide sequence constituting any one of the genes selected from the gene group according to (A) in claim 14. |
| Claim: |
19. An inducer that induces bronchial asthma in a mouse, wherein said inducer comprises as an active ingredient (i) or (ii) in claim 18. |
| Claim: |
20. A method of screening for a therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease comprising the steps of: (1) administering a candidate compound to an animal subject, (2) assaying the expression level of the marker gene in a biological sample obtained from the animal subject, and (3) selecting a compound that decreases the expression level of a marker gene belonging to group (a) or a compound that increases the expression level of a marker gene belonging to group (b), as compared to that in a control with which the candidate compound has not been contacted, wherein the marker gene is any one selected from the group consisting of (a) or (b) in claim 1, or a gene functionally equivalent to said marker gene. |
| Claim: |
21. A method of screening for a therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease comprising the steps of: (1) contacting a candidate compound with a cell into which a vector has been introduced, wherein the vector comprises a transcriptional regulatory region of a marker gene and a reporter gene that is expressed under the control of the transcriptional regulatory region, (2) measuring the activity of the reporter gene, and (3) selecting a compound that decreases the expression level of the reporter gene when the marker gene belongs to group (a), or a compound that increases the expression level of the reporter gene when the marker gene belongs to group (b), as compared to that in a control with which the candidate compound has not been contacted, wherein the marker gene is any one selected from the group according to (a) or (b) in claim 1, or a gene functionally equivalent to the marker gene. |
| Claim: |
22. A method of screening for a therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease comprising the steps of: (1) contacting a candidate compound with a protein encoded by a marker gene, (2) measuring the activity of the protein, and (3) selecting a compound that decreases the activity when the marker gene belongs to group (a), or a compound that increases the activity when the marker gene belongs to the group (b), as compared to that in a control where the candidate compound has not been contacted, wherein the marker gene is any one selected from the group according to (a) or (b) in claim 1, or a gene functionally equivalent to the marker gene. |
| Claim: |
23. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a compound being obtainable by the screening method according to claim 7. |
| Claim: |
24. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a marker gene or an antisense nucleic acid corresponding to a portion of the marker gene, a ribozyme, or a polynucleotide that suppresses the expression of the gene through an RNAi effect, wherein the marker gene is any one selected from the group according to (a) in claim 1. |
| Claim: |
25. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient an antibody recognizing a protein encoded by a marker gene, wherein the marker gene is any one selected from the group according to (a) in claim 1. |
| Claim: |
26. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a marker gene, or a protein encoded by a marker gene, wherein the marker gene is any one selected from the group according to (b) in claim 1. |
| Claim: |
27. A DNA chip for testing for bronchial asthma or a chronic obstructive pulmonary disease, on which a probe has been immobilized to assay a marker gene, and wherein the marker gene comprises at least a single type of gene selected from group (a) and (b) in claim 1. |
| Claim: |
28. The kit according to claim 11, which further comprises a cell-supporting material to culture respiratory epithelial cells under conditions in which the culture medium is supplied from the basolateral side of the cells. |
| Claim: |
29. The kit according to claim 28, which further comprises respiratory epithelial cells. |
| Claim: |
30. A method for producing an animal model for bronchial asthma or chronic obstructive pulmonary disease, which comprises the step of administering to a mouse any one of (i) or (ii): (i) an antisense nucleic acid of a polynucleotide comprising the nucleotide sequence constituting any one of the genes selected from the gene group according to (B) in claim 16, a ribozyme, or a polynucleotide that suppresses the expression of a gene through an RNAi (RNA interference) effect; and (ii) an antibody that binds to a protein encoded by a polynucleotide comprising the nucleotide sequence constituting any one of the genes selected from the gene group according to (B) in claim 16, or a fragment comprising an antigen-binding region thereof. |
| Claim: |
31. An inducer that induces bronchial asthma in a mouse, wherein said inducer comprises as an active ingredient (i) or (ii) in claim 30. |
| Claim: |
32. A method of screening for a therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease comprising the steps of: (1) administering a candidate compound to an animal subject, (2) assaying the expression level of the marker gene in a biological sample obtained from the animal subject, and (3) selecting a compound that decreases the expression level of a marker gene belonging to group (a) or (A), or a compound that increases the expression level of a marker gene belonging to group (b) or (B), as compared to that in a control with which the candidate compound has not been contacted, wherein the marker gene is any one selected from the group consisting of (A) in claim 14, or a gene functionally equivalent to said marker gene. |
| Claim: |
33. A method of screening for a therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease comprising the steps of: (1) administering a candidate compound to an animal subject, (2) assaying the expression level of the marker gene in a biological sample obtained from the animal subject, and (3) selecting a compound that decreases the expression level of a marker gene belonging to group (a) or (A), or a compound that increases the expression level of a marker gene belonging to group (b) or (B), as compared to that in a control with which the candidate compound has not been contacted, wherein the marker gene is any one selected from the group consisting of (B) in claim 16, or a gene functionally equivalent to said marker gene. |
| Claim: |
34. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a compound being obtainable by the screening method according to claim 20. |
| Claim: |
35. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a compound being obtainable by the screening method according to claim 32. |
| Claim: |
36. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a compound being obtainable by the screening method according to claim 33. |
| Claim: |
37. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a compound being obtainable by the screening method according to claim 21. |
| Claim: |
38. A therapeutic agent for bronchial asthma or chronic obstructive pulmonary disease, which comprises as an active ingredient a compound being obtainable by the screening method according to claim 22. |
| Current U.S. Class: |
435006/000 |
| رقم الانضمام: |
edspap.20050208496 |
| قاعدة البيانات: |
USPTO Patent Applications |