Diagnosis and Prediction of Parkinson's Disease

• US 20070277251A1
• Filed: 05/19/2004
• Published: 11/29/2007
• Est. Priority Date: 05/22/2003
• Status: Abandoned Application

• Current Assignee: Licentia Limited
• Original Assignee: Licentia Limited
• Inventors: Wartiovaara, Anu, Luoma, Petri
• Application Number:
  US10/557,649
• Publication Number:
  US 20070277251A1
• Time in Patent Office: Days
• US Class Current: 800/13
• CPC Class Codes: C12N 9/1252 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Y 207/07007 DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Abstract

The present invention provides an in vitro method for diagnosis or prediction of Parkinson'"'"'s disease in a subject, the method comprising testing for a mutation of a mitochondrial DNA polymerase POLG1 gene of the subject and, where the mutation is detected, diagnosing or predicting Parkinson'"'"'s disease in the subject. Further, the present invention provides a diagnostic kit for diagnosis or prediction of Parkinson'"'"'s disease in a subject.

Claims

1. An in vitro method for diagnosis or prediction of Parkinson'"'"'s disease in a subject, the method comprising testing for a mutation of a mitochondrial DNA polymerase POLG1 gene (SEQ ID No. 2) of the subject and, where the mutation is detected, diagnosing or predicting Parkinson'"'"'s disease in the subject.

2. The method according to claim 1, wherein the mutation modifies the function of POLGL protein (SEQ ID No. 1).

3. The method according to claim 1, the method comprising testing for a plurality of said gene mutations.

4. The method according to claim 1, wherein the mutation is tested in a region extending carboxyterminally from the exoIII domain to the spacer and pol regions of the POLG1 protein.

5. The method according to claim 1, wherein the mutation is tested in a DNA location corresponding to nucleotide locations of SEQ ID No. 2 of 7554 to 18478.

6. The method according to claim 1, wherein the mutation is tested in at least one DNA location corresponding to an amino acid location of SEQ ID No. 1 selected from:

467, 468, 627, 953, 955, 1105 and 1236, or in at least one nucleotide location of SEQ ID No. 2 selected from;

8258-8259, 10873, 11381, 11818, 15661, 15686 and 17374.

7. The method according to claim 6, wherein the mutation corresponds to at least one amino acid change selected from:

A467T, N468D, R627Q, R953C, Y955C, A1105T and Q1236H, or at least one nucleotide change selected from;

8258-8259 with an additional G, T10873C, C11381T, T11818C, A15661G, T15686C and C17374A.

8. The method according to claim 1, wherein the mutation is tested in at least one DNA location corresponding to an amino acid location of SEQ ID No. 1 selected from:

43-52, 312, 467, 468, 627,748, 953, 955, 1105, 1143, 1230 and 1236, or at least one nucleotide location of SEQ ID No. 2 selected from;

1168-1197, 8258-8259, 10873, 11381, 11818, 15661, 15686, 15790 and 17374.

9. The method according to claim 8, wherein the mutation corresponds to at least one amino acid change selected from:

A467T, 43-52PolyQ43-50, W312R, N468D, R627Q, W748S, R953C, Y955C, A1105T, E1143G, S1230F and Q1236H, or at least one nucleotide change selected from;

1168-1197 with missing CAGCAG((cag)₈-allele), 8258-8259 with additional G, T10873C, C11381T, T11818C, A15661G, T15686C, G15790A and C17374A.

10. The method according to claim 1, wherein testing is carried out on a nucleic acid sample obtained from the patient.

11. The method according to claim 1, the method comprising testing for a mutant allele, wherein detection of at least one POLG1 mutant allele indicates that the patient has or is predisposed to having Parkinson'"'"'s disease.

12. The method according to claim 1, the method comprising testing for a mutant allele, wherein determination that the subject is homozygous or compound heterozygous for a mutation indicates that the patient has or is predisposed to having Parkinson'"'"'s disease.

13. The method according to claim 1, wherein said testing comprises a procedure selected from:

allele specific oligonucleotide hybridization;

size analysis;

sequencing;

hybridization;

5′

nuclease digestion;

single-stranded conformation polymorphism;

allele specific hybridization;

primer specific extension;

oligonucleotide ligation assay;

temperature gradient electrophoresis;

microarray; and

mass spectrometry.

14. The method according to claim 13, wherein said size analysis is preceded by a restriction enzyme digestion.

15. The method according to claim 10, further comprising amplifying the nucleic acid sample.

16. The method according to claim 15, wherein amplifying the nucleic acid sample employs a primer pair selected from the sequences SEQ ID Nos. 3-11.

17. A diagnostic kit for diagnosis or prediction of Parkinson'"'"'s disease in a subject, comprising means for testing for a mutation of the POLG1 gene of the subject and means for determining whether the mutation is indicative for Parkinson'"'"'s disease.

18. The diagnostic kit according to claim 17, comprising means for amplifying DNA and a labeled polynucleotide comprising a nucleotide sequence complementary to at least part of the gene encoding POLG1 and containing at least one mutation.

19. (canceled)

20. (canceled)

21. (canceled)

22. An isolated polynucleotide comprising at least one mutation of the POLG1 gene, or a fragment of said polynucleotide.

23. The polynucleotide according to claim 22, being labeled.

24. The polynucleotide according to claim 22, wherein the mutation is situated in a region between the C-terminus and the exoIII domain of the POLG1 gene.

25. The polynucleotide according to claim 22, wherein the mutation is situated in a DNA location of SEQ ID No. 2 of 7554 to 18478.

26. The polynucleotide according to claim 22, wherein the mutation is situated in a DNA location corresponding to at least one amino acid location of the SEQ ID No. 1 selected from:

467, 468, 627, 953, 955, 1105 and 1236, or at least one nucleotide location of the SEQ ID No. 2 selected from;

8258-8259, 10873, 11381, 11818, 15661, 15686, 17374.

27. The polynucleotide according to claim 24, wherein the mutation corresponds to at least one amino acid change selected from:

A467T, N468D, R627Q, R953C, Y955C, A1105T and Q1236H, or genomic nucleotide change selected from;

8258-8259 with an additional G, T10873C, C11381T, T11818C, A15661G, T15686C and C17374A.

28. The polynucleotide according to claim 22, wherein the mutation is situated in a DNA location corresponding to at least one amino acid location of SEQ ID No. 1 selected from:

43-52, 312, 467, 468, 627, 748, 953, 955, 1105, 1143, 1230 and 1236, or at least one nucleotide location of SEQ ID No. 2 selected from;

1168-1197, 8258-8259, 10873, 11381, 11818, 15661, 15686, 15790 and 17374.

29. The polynucleotide according to claim 28, wherein the mutation corresponds to at least one amino acid change selected from:

A467T, 43-52PolyQ43-50, W312R, N468D, R627Q, W748S, R953C, Y955C, A1105T, E1143G, S1230F and Q1236H, or at least one nucleotide change selected from;

1168-1197 with missing CAGCAG((cag)₈-allele), 8258-8259 with additional G, T10873C, C11381T, T11818C, A15661G, T15686C, G15790A and C17374A.

30. A recombinant vector comprising the polynucleotide according to claim 22.

31. A cell line comprising the polynucleotide according to claim 22.

32. A transgenic non-human mammal or invertebrate comprising the polynucleotide according to claim 22.