MUTATED ACVR1 FOR DIAGNOSIS AND TREATMENT OF FIBRODYPLASIA OSSIFICANS PROGRESSIVA (FOP)

• US 20090253132A1
• Filed: 04/17/2007
• Published: 10/08/2009
• Est. Priority Date: 04/18/2006
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Shore, Eileen M., Kaplan, Frederick S.
• Granted Patent: US 8,895,711 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• 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 2267/0306 Animal model for genetic diseases
• CPC Class Codes: A01K 2267/0331 Animal model for proliferative diseases
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. 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: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/4713 Autoimmune diseases, e.g. Insulin-dependent diabetes mellitus, multiple sclerosis, rheumathoid arthritis, systemic lupus erythematosus; Autoantigens
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• 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: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/91205 Phosphotransferases in general
• CPC Class Codes: G01N 2800/10 Musculoskeletal or connective tissue disorders
• CPC Class Codes: G01N 2800/2842 Pain, e.g. neuropathic pain, psychogenic pain
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes
• CPC Class Codes: G01N 33/6893 : related to diseases not provided for elsewhere

Abstract

This invention is directed to mutated Activin A type I receptor proteins (ACVR1) and isolated nucleic acids encoding same. The invention also relates to the use of mutated ACVR1 in the diagnosis and treatment of Fibrodysplasia Ossificans Progressiva (FOP).

Claims

1. An isolated nucleic acid encoding a mutated Activin A type I receptor protein (ACVR1), represented by SEQ ID NOs:

21-25 and 27-32 or their combination, wherein the isolated nucleic acid enhances activity of bone morphogenetic protein (BMP) receptor or signaling and is pathognomonic of Fibrodysplasia Ossificans Progressiva (FOP) in a subject.

2. The isolated nucleic acid of claim 1, wherein the nucleic acid has a sequence having at least 90% similarity with the nucleic acid coding sequence of a sequence selected from SEQ ID NOs:

21-25 and 27-32.

3. The isolated nucleic acid of claim 1, wherein the mutation in the mutated Activin A type I receptor protein (ACVR1), is in codon 44C→

G mutation, a codon 590-592 deletion CTT mutation, a codon 617G→

A mutation, a codon 619C→

G mutation, a codon 744G→

C mutation a codon 982G→

T mutation, a codon 983G→

A mutation, a codon 1067G→

A mutation, a codon 1124G→

C mutation, or a combination thereof.

4. The isolated nucleic acid of claim 1, wherein the mutation in the mutated Activin A type I receptor protein (ACVR1), results in a A156G mutation, a replacement of Pro197 and Phe 198 with one Leu residue, a R206H mutation, a Q207E mutation, a R258S mutation, a G328R, a G328W mutation, a G328E mutation, a G356D mutation, a R375P mutation, or a combination thereof on the encoded Activin A type I receptor protein (ACVR1).

5. The isolated nucleic acid of claim 1, wherein the nucleic acid is DNA or RNA.

6. The isolated nucleic acid of claim 1, wherein the nucleic acid is cDNA, genomic DNA, or a PCR product.

7. The isolated nucleic acid of claim 1, wherein the nucleic acid is labeled with a detectable marker.

8. The isolated nucleic acid of claim 7, wherein the detectable marker is a radioactive, colorimetric, luminescent, fluorescent marker, or gold label.

9-13. -13. (canceled)

14. A composition, comprising an antibody, a fragment thereof, or a molecular beacon, wherein said antibody, fragment thereof, or molecular beacon is specifically reactive with a mutated Activin A type I receptor protein (ACVR1), represented by SEQ ID NOs:

21-25 and 27-32 or their combination, or a gene encoding said mutated Activin A type I receptor protein (ACVR1) represented by SEQ ID NOs;

21-25 and 27-32 or their combination.

15-26. -26. (canceled)

27. A method of diagnosing Fibrodysplasia Ossificans Progressiva (FOP) in a subject, comprising:

contacting a biological sample from said subject with an antibody, a fragment thereof, or a molecular beacon, said antibody, fragment thereof, or molecular beacon specifically reactive with a nucleic acid sequence encoding a mutated Activin A type I receptor protein (ACVR1) represented by SEQ ID NOs;

21-25 and 27-32 or their combination, or an amino acid sequence of the mutated Activin A type I receptor protein (ACVR1) represented by SEQ ID NOs;

21-25 and 27-32 or their combination; and

assaying for the presence of said mutated Activin A type I receptor protein (ACVR1), wherein the presence of the mutated Activin A type I receptor protein (ACVR1) in said sample indicates that said subject has Fibrodysplasia Ossificans Progressiva (FOP).

28. The method of claim 27, whereby the Fibrodysplasia Ossificans Progressiva (FOP) is inherited Fibrodysplasia Ossificans Progressiva (FOP).

29. The method of claim 27, whereby the Fibrodysplasia Ossificans Progressiva (FOP) is sporadic Fibrodysplasia Ossificans Progressiva (FOP).

30. The method of claim 27, whereby said mutated Activin A type T receptor protein (ACVR1) has a A156G mutation, a replacement of Pro197 and Phe 198 with one Leu residue, a R206H mutation, a Q207E mutation, a R258S mutation, a G328R, a G328W mutation, a G328E mutation, a G356D mutation, a R375P mutation, or a combination thereof.

31. The method of claim 27, whereby said antibody, fragment thereof, or molecular beacon further comprises a detectable label operably linked thereto.

33. The method of claim 27, whereby said fragment is Fe, Fab, F(ab′

)2, scFv or a combination thereof.

34. The method of claim 27, whereby said molecular beacon comprises:

an oligonucleotide comprising a stem and a loop structure and having a photoluminescent dye at one of the 5‘

or J’

ends and a quenching agent at the opposite 3′

or 5′

ends, whereby said loop consists of about 12-25 bases specifically reactive with a gene sequence encoding said mutated Activin A type I receptor protein (ACVR1).

35. The method of claim 27, whereby said step of contacting further comprises amplifying the target sequence of said molecular beacon.

36. The method of claim 27, whereby said antibody fragment thereof, or molecular beacon specifically reactive with the nucleic acid coding sequence of SEQ ID NOs:

21-25 and 27-32, or their combination.

32. (canceled)

37-53. -53. (canceled)

54. A kit for diagnosing Fibrodysplasia Ossificans Progressiva (FOP) in a subject, comprising:

an antibody, a fragment thereof, or a molecular beacon, said antibody, fragment thereof, or molecular beacon specifically reactive with a mutated Activin A type I receptor protein (ACVR1) represented by SEQ ID NOs;

21-25 and 27-32 or their combination, or a gene encoding said mutated Activin A type I receptor protein (ACVR1).

55-68. -68. (canceled)