NUCLEIC ACID ARRAYS FOR DETECTING GENE EXPRESSION IN ANIMAL MODELS OF INFLAMMATORY DISEASES

• US 20090062131A1
• Filed: 05/05/2008
• Published: 03/05/2009
• Est. Priority Date: 10/02/2003
• Status: Abandoned Application

• Current Assignee: Wyeth (Pfizer Inc.)
• Original Assignee: Wyeth (Pfizer Inc.)
• Inventors: MOUNTS, William Martin
• Application Number:
  US12/114,963
• Publication Number:
  US 20090062131A1
• Time in Patent Office: Days
• US Class Current: 506/7
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

The present invention provides nucleic acid arrays and methods of using the same for detecting gene expression in animal models of osteoarthritis or other inflammatory diseases. The nucleic acid arrays of the present invention comprise polynucleotide probes for genes that are differentially expressed in osteoarthritis-affected cartilage tissues as compared to non-osteoarthritic cartilage tissues. In one embodiment, a nucleic acid array of the present invention comprises a plurality of polynucleotide probe sets, each of which is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective tiling sequence selected from Table C, or the complement thereof.

Claims

1. A nucleic acid array comprising at least one polynucleotide probe capable of hybridizing under stringent or nucleic acid array hybridization conditions to a gene which is differentially expressed in osteoarthritic cartilage cells relative to non-osteoarthritic cartilage cells, and wherein said osteoarthritic and non-osteoarthritic cartilage cells are derived from the same species.

2. The nucleic acid array according to claim 1, comprising at least 10 polynucleotide probe sets, wherein each said probe set is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective gene which is differentially expressed in said osteoarthritic cartilage cells relative to said non-osteoarthritic cartilage cells.

3. The nucleic acid array according to claim 1, comprising at least 100 polynucleotide probe sets, wherein each said probe set is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective gene which is differentially expressed in said osteoarthritic cartilage cells relative to said non-osteoarthritic cartilage cells.

4. The nucleic acid array according to claim 3, wherein each said probe set comprises at least 12 polynucleotide probes.

5. The nucleic acid array according to claim 3, wherein the average expression level of each said gene in said osteoarthritic cartilage cells is substantially higher than that in said non-osteoarthritic cartilage cells.

6. The nucleic acid array according to claim 5, further comprising at least 100 polynucleotide probe sets, each of which is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective gene whose average expression level in said non-osteoarthritic cartilage cells is substantially higher than that in said osteoarthritic cartilage cells.

7. The nucleic acid array according to claim 6, wherein said osteoarthritic and non-osteoarthritic cartilage cells are Canis familiaris cartilage cells.

8. The nucleic acid array according to claim 1, comprising at least one polynucleotide probe which is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a tiling sequence selected from Table C, or the complement thereof, wherein said tiling sequence has a pattern value of 010, 011, 100 or 101, as shown in Table D.

9. The nucleic acid array according to claim 1, comprising at least 10 polynucleotide probe sets, wherein each said probe set is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective tiling sequence selected from Table C, or the complement thereof, and wherein each said tiling sequence has a pattern value of 010, 011, 100 or 101, as shown in Table D.

10. The nucleic acid array according to claim 1, comprising at least 100 polynucleotide probe sets, wherein each said probe set is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective tiling sequence selected from Table C, or the complement thereof, and wherein each said tiling sequence has a pattern value of 010, 011, 100 or 101, as shown in Table D.

11. A method of screening for drug candidates capable of modulating expression of genes that are differentially expressed in osteoarthritic cartilage cells relative to non-osteoarthritic cartilage cells, comprising the steps of:

(a) preparing a first nucleic acid sample from a vertebrate affected by osteoarthritis;

(b) hybridizing the first nucleic acid sample to a first nucleic acid array as in any one of claims 1-4;

(c) detecting a first set of hybridization signals;

(d) treating the vertebrate with a candidate drug;

(e) repeating steps (a)-(c) with a second nucleic acid sample from the treated vertebrate and a second nucleic acid array identical to the first array to obtain a second set of hybridization signals; and

(f) comparing the first and second sets of hybridization signals,

wherein any change in expression level of at least one gene differentially expressed in osteoarthritic cartilage cells relative to non-osteoarthritic cartilage cells identifies the candidate drug as one that modulates expression of said gene.

12. The method according to claim 11, wherein the vertebrate is a canine animal, and the first and second nucleic acid samples are prepared from cartilage tissues of said canine animal.

13. A method of screening for drug candidates capable of modulating expression of genes that are differentially expressed in osteoarthritic cartilage cells relative to non-osteoarthritic cartilage cells, comprising the steps of:

(a) preparing a first nucleic acid sample from a cartilage cell or tissue affected by osteoarthritis;

(b) hybridizing the first nucleic acid sample to a first nucleic acid array as in any one of claims 1-4;

(c) detecting a first set of hybridization signals;

(d) treating the cell or tissue with a candidate drug;

(e) repeating steps (a)-(c) with a second nucleic acid sample from the treated cell or tissue and a second nucleic acid array identical to the first array to obtain a second set of hybridization signals; and

(f) comparing the first and second sets of hybridization signals,

wherein any change in expression level of at least one gene differentially expressed in osteoarthritic cartilage cells relative to non-osteoarthritic cartilage cells identifies the candidate drug as one that modulates expression of said gene.

14. A method for detecting gene expression in a sample of interest, comprising:

hybridizing nucleic acid molecules prepared from said sample to a nucleic acid array as in any one of claims 1-4; and

detecting hybridization signals on the nucleic acid array.

15. A nucleic acid array comprising a plurality of polynucleotide probes, wherein each said probe is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective tiling sequence selected from Table C, or the complement thereof.

16. The nucleic acid array according to claim 15, wherein said plurality of polynucleotide probes includes at least 10 probe sets, and each said probe set is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective tiling sequence selected from Table C, or the complement thereof.

17. The nucleic acid array according to claim 15, wherein said plurality of polynucleotide probes includes at least 100 probe sets, and each said probe set is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective tiling sequence selected from Table C, or the complement thereof.

18. A method of making a nucleic acid array, comprising the steps of:

selecting a plurality of polynucleotide probes, each of which is capable of hybridizing under stringent or nucleic acid array hybridization conditions to a different respective gene which is differentially expressed in osteoarthritic cartilage cells relative to non-osteoarthritic cartilage cells; and

attaching the plurality of polynucleotide probes to one or more substrate supports, wherein said osteoarthritic and non-osteoarthritic cartilage cells are derived from the same species.

19. A polynucleotide collection comprising at least one polynucleotide capable of hybridizing under stringent or nucleic acid array hybridization conditions to a parent sequence selected from SEQ ID NOs:

1-12,167, or the complement thereof.

20. A probe array comprising a plurality of probes capable of binding to expression products of genes that are differentially expressed in osteoarthritic cartilage cells relative to non-osteoarthritic cartilage cells, wherein said osteoarthritic cells and said non-osteoarthritic-cartilage cells are derived from the same species.

21. The probe array according to claim 20, wherein each said gene encodes a tiling sequence selected from Table C and having a pattern value of 010, 011, 100 or 101.