Promoter Variants Of The Alpha-7 Nicotinic Acetylcholine Receptor

• US 20100047795A1
• Filed: 05/29/2009
• Published: 02/25/2010
• Est. Priority Date: 10/23/1997
• Status: Active Grant

• Current Assignee: US Department of Veterans Affairs (Government of the United States of America)
• Original Assignee: US Department of Veterans Affairs (Government of the United States of America)
• Inventors: Freedman, Robert, Leonard, Sherry
• Granted Patent: US 8,236,500 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/70571 for neuromediators, e.g. serotonin receptor, dopamine receptor
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: G01N 2333/705 Assays involving receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: G01N 2800/302 Schizophrenia
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The present invention is directed to methods and compositions related to α7 acetylcholine nicotinic receptor genes, in particular, the human α7 nicotinic acetylcholine receptor gene. The human α7 nicotinic acetylcholine receptor gene is associated with the pathophysiological aspects of the disease schizophrenia. The present invention further provides methods and compositions to screen populations for abnormal α7 alleles, as well as methods and compositions for development of therapeutics.

Claims

1. A composition comprising an isolated 5′

upstream regulatory nucleotide sequence of a human alpha-7 nicotinic receptor, wherein said nucleotide sequence comprises at least one polymorphism.

2. The composition of claim 1, wherein said sequence comprises at least a portion of SEQ ID NO:

181.

3. The composition of claim 1, wherein said at least one polymorphism comprises one or more of those listed in Table 23.

4. The composition of claim 1, wherein said polymorphism is rs3087454 (−

1831 bp).

5. A composition comprising an isolated regulatory binding site sequence encoded by an isolated 5′

upstream regulatory nucleotide sequence of a human alpha-7 nicotinic receptor, wherein said nucleotide sequence comprises at least one polymorphism.

6. The composition of claim 5, wherein said sequence comprises at least a portion of SEQ ID NO:

181.

7. The composition of claim 5, wherein said at least one polymorphism comprises one or more of those listed in Table 23.

8. The composition of claim 5, wherein said polymorphism is rs3087454 (−

1831 bp).

9. A method, comprising:

a) providing;

i) an individual suspected of having a predisposition to schizophrenia;

ii) a nucleic acid derived from said individual, wherein said nucleic acid comprises an α

7 nicotinic acid receptor regulatory allele;

b) detecting at least one polymorphism within said α

7 regulatory allele; and

c) correlating the presence of said at least one polymorphism with a predisposition to schizophrenia.

10. The method of claim 9, wherein said at least one polymorphism comprises one or more of those listed in Table 23.

11. The method of claim 9, wherein said polymorphism is rs3087454 (−

1831 bp).

12. The method of claim 10, wherein said at least one polymorphism comprises two or more polymorphisms.

13. The method of claim 9, wherein said at least one polymorphism contributes to reduced α

7 regulatory protein transcription.

14. The method of claim 9, wherein said detecting comprises at least one technique selected from the group consisting of polymerase chain reaction, heteroduplex analysis, single stand conformational polymorphism analysis, denaturing high performance liquid chromatography, ligase chain reaction, comparative genome hybridisation, Southern blotting, and sequencing.

15. The method of claim 9, wherein said nucleic acid is derived from a biological sample selected from the group consisting of a biopsy material and blood.

16. The method of claim 9, further comprising step (d) providing a diagnosis to said individual based on the presence or absence of the at least one polymorphism.

17. The method of claim 9, wherein said diagnosis differentiates schizophrenia from other forms of mental illness.

18. A method, comprising:

a) providing;

i) a biological sample suspected of containing a first polynucleotide encoding an α

7 regulatory protein;

ii) a second polynucleotide comprising at least a portion of SEQ ID NO;

181, capable of hybridizing to said first polynucleotide;

b) hybridizing said first polynucleotide to said second polynucleotide to produce a hybridization complex;

c) detecting an α

7 nicotinic acid regulatory allele within said first polynucleotide, wherein said allele comprises at least one polymorphism.

19. The method of claim 18, wherein said at least one polymorphism is rs3087454 (−

1831 bp).

20. The method of claim 18, wherein at least one polymorphism comprises one or more of those listed in Table 23.

21. The method of claim 18, wherein said method further comprises step (d) detecting said hybridization complex, wherein said complex correlates with said first polynucleotide.

22. The method of claim 18, wherein said biological sample is selected from the group consisting of brain tissue and blood.

23. The method of claim 18, wherein said biological sample is from a human.

24. The method of claim 23, wherein said human is suspected of suffering from a condition selected from the group consisting of schizophrenia, small cell lung carcinoma, breast cancer, and nicotine-dependent illness.

25. The method of claim 18, wherein said method further comprises, before step (b) amplifying said first polynucleotide by polymerase chain reaction.