Growth differentiation factor receptors, agonists and antagonists thereof, and methods of using same

• US 6,656,475 B1
• Filed: 07/27/2000
• Issued: 12/02/2003
• Est. Priority Date: 08/01/1997
• Status: Expired due to Term

• Current Assignee: Johns Hopkins University
• Original Assignee: Johns Hopkins University
• Inventors: McPherron, Alexandra C., Lee, Se-Jin
• Primary Examiner(s): Eyler, Yvonne
• Assistant Examiner(s): Andres, Janet L.
• Application Number:
  US09/626,896
• Time in Patent Office: 1,223 Days
• Field of Search: 514/2, 424/198.1, 530/350
• US Class Current: 424/198.1
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 2267/0306 Animal model for genetic diseases
• CPC Class Codes: A01K 2267/0362 Animal model for lipid/glucose metabolism, e.g. obesity, type-2 diabetes
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A61P 1/14 Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 : cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 2830/85 : mammalian
• CPC Class Codes: G01N 2333/495 : Transforming growth factor [TGF]
• CPC Class Codes: G01N 2500/00 : Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/74 : involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

The present invention provides a substantially purified growth differentiation factor (GDF) receptor, including a GDF-8 (myostatin) receptor, as well as functional peptide portions thereof. In addition, the invention provides a virtual representation of a GDF receptor or a functional peptide portion thereof. The present invention also provides a method of modulating an effect of myostatin on a cell by contacting the cell with an agent that affects myostatin signal transduction in the cell. In addition, the invention provides a method of ameliorating the severity of a pathologic condition, which is characterized, at least in part, by an abnormal amount, development or metabolic activity of muscle or adipose tissue in a subject, by modulating myostatin signal transduction in a muscle cell or an adipose tissue cell in the subject. The invention also provides a method of modulating the growth of muscle tissue or adipose tissue in a eukaryotic organism by administering an agent that affects myostatin signal transduction to the organism.

Claims

1. A method of reducing or inhibiting an effect of myostatin on a cell, comprising contacting the cell with a myostatin prodomain, which alters a specific interaction of myostatin with a myostatin receptor expressed by the cell, said myostatin prodomain consisting of about amino acid residues 20 to 262 of a promyostatin polypeptide selected from the group consisting of SEQ ID NOS:

2, 4, 6, 8, 10, 12, 14, 16, 18 and 20, thereby modulating the effect of myostatin on the cell.

2. The method of claim 1, wherein the myostatin receptor is an activin receptor.

3. The method of claim 2, wherein the activin receptor is an activin type IIB receptor.

4. The method of claim 1, wherein the myostatin prodomain consisting of

amino acid sequence set forth as

amino acid residues about 20 to 263 as set forth in SEQ ID NO:

4;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

2;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

10;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

12;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

8;

amino acid residues about 20 to 263 as set forth in SEQ ID NO;

6;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

18;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

14;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

16;

or

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

20.

5. A method of ameliorating the severity of a pathologic condition characterized, at least in part, by an abnormal amount, development or metabolic activity of muscle in a subject, said method comprising contacting a muscle cell in the subject with a myostatin prodomain, said myostatin prodomain consisting of about amino acid residues 20 to 262 of a promyostatin polypeptide as set forth in any of SEQ ID NOS:

2, 4, 6, 8, 10, 12, 14, 16, 18 and 20, thereby modulating myostatin signal transduction in the muscle cell in the subject and ameliorating the severity of the pathologic condition.

6. The method of claim 5, wherein the pathological condition is a wasting disorder, said method comprising reducing or inhibiting myostatin signal transduction in a muscle cell.

7. The method of claim 6, wherein the wasting disorder is selected from the group consisting of cachexia and anorexia.

8. The method of claim 6, wherein the wasting disorder is a muscle wasting disorder selected from the group consisting of a muscular dystrophy and a neuromuscular disease.

9. The method of claim 5, wherein the pathological condition is a metabolic disorder.

10. The method of claim 9, wherein the metabolic disorder is selected from the group consisting of obesity and type II diabetes.

11. The method of claim 10, comprising reducing or inhibiting myostatin signal transduction in a muscle cell.

22. The method of claim 5, wherein the myostatin prodomain consisting of an amino acid sequence set forth as

amino acid residues about 20 to 263 as set forth in SEQ ID NO:

4;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

2;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

10;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

12;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

8;

amino acid residues about 20 to 263 as set forth in SEQ ID NO;

6;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

18;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

14;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

16;

or

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

20.

12. A method of increasing the growth of muscle tissue in a eukaryotic organism, comprising administering to the organism a myostatin prodomain, said myostatin prodomain consisting of about amino acid residues 20 to 262 of a promyostatin polypeptide as set forth in any of SEQ ID NOS:

2, 4, 6, 8, 10, 12, 14, 16, 18 and 20, thereby modulating the growth of the muscle tissue in the eukaryotic organism.

13. The method of claim 12, wherein the myostatin prodomain affects myostatin signal transduction, thereby modulating the growth of muscle tissue or adipose tissue in the organism.

14. The method of claim 13, wherein the myostatin prodomain alters the specific interaction of myostatin with a myostatin receptor, thereby affecting myostatin signal transduction.

15. The method of claim 13, wherein the myostatin prodomain reduces or inhibits the specific interaction of myostatin with a myostatin receptor, thereby increasing the growth of the organism.

16. The method of claim 12, wherein the eukaryotic organism is a vertebrate organism.

17. The method of claim 16, wherein the vertebrate organism is selected from the group consisting of a bovine, ovine, porcine, canine, feline, murine, and piscine organism.

18. The method of claim 12, wherein the eukaryotic organism is an invertebrate organism.

19. The method of claim 18, wherein the invertebrate organism is a mollusk.

20. The method of claim 19, comprising administering to the mollusk an agent that reduces or inhibits the specific interaction of myostatin with a myostatin receptor, thereby increasing the growth of the mollusk.

21. The method of claim 19, wherein the mollusk is selected from the group consisting of a shrimp, a scallop, a squid, an octopus, and a snail.

23. The method of claim 12, wherein the myostatin prodomain consisting of an amino acid sequence set forth as

amino acid residues about 20 to 263 as set forth in SEQ ID NO:

4;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

2;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

10;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

12;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

8;

amino acid residues about 20 to 263 as set forth in SEQ ID NO;

6;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

18;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

14;

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

16;

or

amino acid residues about 20 to 262 as set forth in SEQ ID NO;

20.