Chromosome 13-linked breast cancer susceptibility gene

• US 5,837,492 A
• Filed: 04/29/1996
• Issued: 11/17/1998
• Est. Priority Date: 12/18/1995
• Status: Expired due to Term

• Current Assignee: Endorecherche, HSC Research and Development Limited Partnership, Pennsylvania University of The Trustees of The, University of Utah Research Foundation (State of Utah)
• Original Assignee: Endorecherche, HSC Research and Development Limited Partnership, Myriad Genetics, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Tavtigian, Sean V., Rommens, Johanna M., Kamb, Alexander, Simard, Jacques, Weber, Barbara L., Couch, Fergus
• Primary Examiner(s): Chambers, Jasemine C.
• Assistant Examiner(s): HAUDA, KAREN MARIE
• Application Number:
  US08/639,501
• Time in Patent Office: 932 Days
• Field of Search: 435/320.1, 435/375, 435/6, 435/172.3, 435/69.1, 530/828, 530/350, 935/62, 935/52, 935/34, 935/71, 935/70, 935/33, 935/65
• US Class Current: 435/69.1
• 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 14/4703 Inhibitors; Suppressors
• CPC Class Codes: Y10S 530/828 Cancer

Abstract

The present invention relates generally to the field of human genetics. Specifically, the present invention relates to methods and materials used to isolate and detect a human breast cancer predisposing gene (BRCA2), some mutant alleles of which cause susceptibility to cancer, in particular breast cancer. More specifically, the invention relates to germline mutations in the BRCA2 gene and their use in the diagnosis of predisposition to breast cancer. The present invention further relates to somatic mutations in the BRCA2 gene in human breast cancer and their use in the diagnosis and prognosis of human breast cancer. Additionally, the invention relates to somatic mutations in the BRCA2 gene in other human cancers and their use in the diagnosis and prognosis of human cancers. The invention also relates to the therapy of human cancers which have a mutation in the BRCA2 gene, including gene therapy, protein replacement therapy and protein mimetics. The invention further relates to the screening of drugs for cancer therapy. Finally, the invention relates to the screening of the BRCA2 gene for mutations, which are useful for diagnosing the predisposition to breast cancer.

Claims

1. An isolated DNA molecule coding for a BRCA2 polypeptide, said DNA molecule comprising a nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO:

2.

2. The isolated DNA molecule of claim 1, wherein said DNA molecule comprises the nucleotide sequence set forth in SEQ ID NO:

1.

3. The isolated DNA molecule of claim 1, wherein said DNA molecule is an allelic variant of the nucleotide sequence set forth in SEQ ID NO:

1.

4. The isolated DNA molecule of claim 1, which contains BRCA2 regulatory sequences.

5. An isolated DNA molecule comprising at least 15 contiguous nucleotides of the DNA molecule of claim 1.

11. A replicative cloning vector which comprises the isolated DNA molecule of claim 1, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 1, and a replicon operative in a host cell.

12. A replicative cloning vector which comprises the isolated DNA molecule of claim 2, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 2, and a replicon operative in a host cell.

13. A replicative cloning vector which comprises the isolated DNA molecule of claim 3, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 3, and a replicon operative in a host cell.

16. An expression vector which comprises the isolated DNA of claim 1, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 1, operably linked to transcription regulatory regions.

17. An expression vector which comprises the isolated DNA of claim 2, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 2, operably linked to transcription regulatory regions.

18. An expression vector which comprises the isolated DNA of claim 3, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 3, operably linked to transcription regulatory regions.

21. An isolated host cell transformed with the expression vector of claim 16.

22. An isolated host cell transformed with the expression vector of claim 17.

23. An isolated host cell transformed with the expression vector of claim 18.

26. A method of producing recombinant BRCA2 polypeptide which comprises culturing the cells of claim 21 under conditions effective for the production of said BRCA2 polypeptide and harvesting the recombinant BRCA2 polypeptide.

27. A method of producing recombinant BRCA2 polypeptide which comprises culturing the cells of claim 22 under conditions effective for the production of said BRCA2 polypeptide and harvesting the recombinant BRCA2 polypeptide.

28. A method of producing recombinant BRCA2 polypeptide which comprises culturing the cells of claim 23 under conditions effective for the production of said BRCA2 polypeptide and harvesting the recombinant BRCA2 polypeptide.

6. An isolated DNA molecule coding for a mutated form of the BRCA2 polypeptide set forth in SEQ ID NO:

2, wherein said mutated form of the BRCA2 polypeptide is associated with susceptibility to cancer.

7. The isolated DNA molecule of claim 6, wherein the DNA molecule comprises a mutated nucleotide sequence set forth in SEQ ID NO:

1.

8. The isolated DNA molecule of claim 7, wherein the mutation is selected from the group consisting of a deletion mutation, a nonsense mutation, an insertion mutation and a missense mutation.

9. An isolated DNA molecule comprising at least 15 contiguous nucleotides of the DNA of claim 6.

10. The isolated DNA molecule of claim 6 selected from the group consisting of:

(a) SEQ ID NO;

1 having AC at nucleotide positions 277 and 278 deleted;

(b) SEQ ID NO;

1 having four nucleotides at positions 982-985 deleted;

(c) SEQ ID NO;

1 having four nucleotides at positions 4706-4709 deleted;

(d) SEQ ID NO;

1 having C at nucleotide position 8525 deleted;

(e) SEQ ID NO;

1 having five nucleotides at positions 9254-9258 deleted;

(f) SEQ ID NO;

1 having GT at nucleotide positions 4075 and 4076 deleted;

(g) SEQ ID NO;

1 having five nucleotides at positions 999-1003 deleted;

(h) SEQ ID NO;

1 having T at nucleotide position 6174 deleted;

(i) SEQ ID NO;

1 having three nucleotides at positions 4132-4134 deleted;

(j) SEQ ID NO;

1 having a C instead of a G at position 451;

(k) SEQ ID NO;

1 having a C instead of an A at position 1093;

(l) SEQ ID NO;

1 having a C instead of a G at position 1291;

(m) SEQ ID NO;

1 having A at position 1493 deleted;

(n) SEQ ID NO;

1 having a T instead of a C at position 2117;

(o) SEQ ID NO;

1 having a C instead of an A at position 2411;

(p) SEQ ID NO;

1 having an A instead of a G at position 4813;

(q) SEQ ID NO;

1 having a G instead of a T at position 5868;

(r) SEQ ID NO;

1 having a T instead of a C at position 5972;

(s) SEQ ID NO;

1 having a T instead of a C at position 6328;

(t) SEQ ID NO;

1 having a T instead of a G at position 7049;

(u) SEQ ID NO;

1 having a C instead of a G at position 7491;

(v) SEQ ID NO;

1 having a G instead of an A at position 9537;

(w) SEQ ID NO;

1 having a T instead of an A at position 10204;

(x) SEQ ID NO;

1 having a G instead of a C at position 10298;

(y) SEQ ID NO;

1 having a G instead of an A at position 10462;

(z) SEQ ID NO;

1 having an A instead of a G at position 203;

(aa) SEQ ID NO;

1 having an A instead of a C at position 1342;

(bb) SEQ ID NO;

1 having a C instead of a T at position 2457;

(cc) SEQ ID NO;

1 having a G instead of an A at position 3199;

(dd) SEQ ID NO;

1 having a G instead of an A at position 3624;

(ee) SEQ ID NO;

1 having a G instead of an A at position 3668;

(ff) SEQ ID NO;

1 having a C instead of a T at position 4035;

(gg) SEQ ID NO;

1 having a G instead of an A at position 7470;

(hh) SEQ ID NO;

1 having a G instead of an A at position 1593;

(ii) SEQ ID NO;

1 having an A instead of a G at position 4296;

(jj) SEQ ID NO;

1 having a G instead of an A at position 5691;

(kk) SEQ ID NO;

1 having a G instead of an A at position 6051;

(ll) SEQ ID NO;

1 having a C instead of a T at position 6828; and

(mm) SEQ ID NO;

1 having a C instead of a T at position 6921.

14. A replicative cloning vector which comprises the isolated DNA molecule of claim 6, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 6, and a replicon operative in a host cell.

15. A replicative cloning vector which comprises the isolated DNA molecule of claim 7, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 7, and a replicon operative in a host cell.

19. An expression vector which comprises the isolated DNA of claim 6, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 6, operably linked to transcription regulatory regions.

20. An expression vector which comprises the isolated DNA of claim 7, or at least 15 contiguous nucleotides of the isolated DNA molecule of claim 7, operably linked to transcription regulatory regions.

24. An isolated host cell transformed with the expression vector of claim 19.

25. An isolated host cell transformed with the expression vector of claim 20.

29. A pair of single-stranded DNA primers of at least 15 nucleotides in length for determination of the nucleotide sequence of a BRCA2 gene by a polymerase chain reaction, the sequence of said primers being isolated from human chromosome 13, wherein the use of said primers in a polymerase chain reaction results in the synthesis of DNA comprising all or at least 15 contiguous nucleotides of the BRCA2 gene.

30. The pair of primers of claim 29 wherein said BRCA2 gene has the nucleotide sequence set forth in SEQ ID NO:

1.