Mutant human hedgehog gene

• US 5,759,811 A
• Filed: 11/13/1996
• Issued: 06/02/1998
• Est. Priority Date: 11/13/1996
• Status: Expired due to Fees

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Hu, Zhilan, Bonifas, Jeanette, Epstein, Ervin
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Stole, Einar
• Application Number:
  US08/748,591
• Time in Patent Office: 566 Days
• Field of Search: 435/69.1, 435/320.1, 530/350, 536/23.1, 935/22
• US Class Current: 435/69.1
• 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: C07K 14/4702 Regulators; Modulating activity

Abstract

A mutation in the human sonic hedgehog gene is associated with tumorigenesis. A variety of human tumors, including basal cell carcinomas, breast carcinomas, medulloblastomas, etc., have a somatic mutation that results in an amino acid substitution at position 133 *his133 SHH!, or in a mutation at position 114. Such mutated genes and fragments thereof, encoded protein, and antibodies specific for the mutated protein are useful in characterizing the phenotype of associated tumors. The mutant protein is useful in drug screening for compositions that antagonize or otherwise modulate HH activity or expression. The encoded protein is also used as a therapeutic, to modulate cell proliferation and differentiation, and treatment of pathological conditions associated with decreased hedgehog signaling.

Claims

1. An isolated nucleic acid other than an intact chromosome encoding an oncogenic human hedgehog (HH) protein that comprises an amino acid substitution or deletion at the position corresponding to position 27 in any one of SEQ ID NO:

1, SEQ ID NO 2 or SEQ ID NO;

3;

or a fragment of at least 18 nucleotides derived therefrom and encompassing said amino acid substitution or deletion.

5. An expression cassette comprising a transcriptional initiation region, a nucleic acid according to claim 1 under the transcriptional regulation of said transcriptional initiation region, and a transcriptional termination region.

6. A cell comprising an expression cassette according to claim 5.

7. A cell according to claim 6, wherein said cell is a mammalian cell.

8. A cell according to claim 7, wherein said mammalian cell is human.

9. A method for producing oncogenic HH protein, said method comprising:

growing a cell according to claim 6, whereby said oncogenic HH protein is expressed, and

isolating said oncogenic HH protein free of other proteins.

15. An isolated nucleic acid according to claim 1, wherein said amino acid substitution comprises a substitution of an amino acid selected from the group consisting of tyrosine phenylalanine or tryptophan.

2. An isolated nucleic acid other than an intact chromosome encoding an oncogenic human hedgehog (HH) protein that comprises an amino acid substitution or deletion at the position corresponding to position 10 in any one of SEQ ID NO:

6, SEQ ID NO;

7 or SEQ ID NO;

8;

or a fragment of at least 18 nucleotides derived therefrom and encompassing said amino acid substitution or deletion.

10. An expression cassette comprising a transcriptional initiation region, a nucleic acid according to claim 2 under the transcriptional regulation of said transcriptional initiation region, and a transcriptional termination region.

11. A cell comprising an expression cassette according to claim 10.

12. A cell according to claim 11, wherein said cell is a mammalian cell.

13. A cell according to claim 12, wherein said mammalian cell is human.

14. A method for producing oncogenic HH protein, said method comprising:

growin a cell according to claim 11, whereby said oncogenic HH protein is expressed, and

isolating said oncogenic HH protein free of other proteins.

3. An isolated nucleic acid other than an intact chromosome encoding an oncogenic hedgehog (HR) protein selected from the group consisting of human his→

tyr HH! and met→

ile HH!;

or a fragment of at least 18 nucleotides derived therefrom and encompassing said his→

try HH! or met→

ile HH! mutation.

4. An isolated nucleic acid according to claim 3, selected from the group consisting of SEQ ID NO:

5 and SEQ ID NO;

10.

16. An isolated nucleic acid of at least 18 nucleotides of contiguous sequence as set forth in SEQ ID NO:

5, and encompassing the his→

tyr HH! mutation.

17. An isolated nucleic acid of at least 18 nucleotides of contiguous sequence as set forth in SEQ ID NO:

10, and encompassing the met→

ile HH! mutation.