Mutated ACVR1 for diagnosis and treatment of Fibrodyplasia Ossificans Progressiva (FOP)

• US 10,106,852 B2
• Filed: 10/22/2014
• Issued: 10/23/2018
• 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: Kaplan, Frederick S., Shore, Eileen M.
• Primary Examiner(s): Kapushoc, Stephen T
• Application Number:
  US14/521,143
• Publication Number:
  US 20150203918A1
• Time in Patent Office: 1,462 Days
• 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. A nucleic acid comprising a a double stranded DNA duplex comprising two complementary strands of DNA, wherein one of the two complementary strands of DNA contiguously encodes SEQ ID NO:

21 or a fragment thereof comprising SEQ ID NO;

42.

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

3. The nucleic acid of claim 2, wherein the detectable marker is a radioactive, colorimetric, luminescent, fluorescent marker, a photoluminescent dye, or a gold label.

23. The nucleic acid of claim 1, wherein one of the two complementary strands of DNA contiguously encodes SEQ ID NO:

21.

4. A composition comprising a molecular beacon, said molecular beacon comprising:

an oligonucleotide comprising a stem and a loop, and a detectable label, wherein the loop comprises at least 12 contiguous bases of SEQ ID NO;

34 or its complement, and wherein position 11 of SEQ ID NO;

34 is an A.

5. The composition of claim 4, wherein the oligonucleotide of said molecular beacon comprises a photoluminescent dye at its 5′

or 3′

end and a quenching agent at the opposite 3′

or 5′

end respectively, wherein said loop is 12-25 bases in length.

6. The composition of claim 5, wherein said photoluminescent dye is fluorescein, phycoerythrin, CY3, CY5, allophycocyanine, Texas Red, peridenin chlorophyll, cyanine, FAM, JOE, TAMRA, TET, VIC, or a combination thereof.

7. The composition of claim 5, wherein said quenching agent is EDANS, BHQ1, DABCYL, BODIPY FL, BH1, BH2, QSY7, or a combination thereof.

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

contacting a biological sample from said subject with a molecular beacon of claim 4; and

assaying for the presence of a mutated Activin A type I receptor protein (ACVR1) that encodes histidine at the position corresponding to position 206 of SEQ ID NO;

21, 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).

9. The method of claim 8, whereby the Fibrodysplasia Ossificans Progressiva (FOP) is inherited Fibrodysplasia Ossificans Progressiva (FOP).

10. The method of claim 8, whereby the Fibrodysplasia Ossificans Progressiva (FOP) is sporadic Fibrodysplasia Ossificans Progressiva (FOP).

11. The method of claim 8, whereby said molecular beacon further comprises a detectable label operably linked thereto.

12. The method of claim 11, whereby said label is an enzyme.

13. The method of claim 8, 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 3′

ends and a quenching agent at the opposite 3′

or 5′

ends.

14. The method of claim 8, further comprising amplifying a target sequence of said molecular beacon.

15. The method of claim 11, whereby said label is a radiolabel, a fluorophore, a peptide, an enzyme, a quantum dot, or a combination thereof.

16. The method of claim 15, whereby said quantum dot is CdSe.

17. The method of claim 13, whereby said photoluminescent dye is fluorescein, phycoerythrin, CY3, CY5, allophycocyanine, Texas Red, peridenin chlorophyll, cyanine, FAM, JOE, TAMRA, TET, VIC, or a combination thereof.

18. The method of claim 13, whereby said quenching agent is EDANS, BHQ1, DABCYL, BODIPY FL, BH1, BH2, QSY7, or a combination thereof.

19. A kit for diagnosing Fibrodysplasia Ossificans Progressiva (FOP) in a subject, comprising the composition of claim 4.

20. The kit of claim 19, wherein the detectable label is a radiolabel, a fluorophore, a peptide, an enzyme, a quantum dot, or a combination thereof.

21. The kit of claim 19, further comprising at least one standard, obtained from a subject, or pool of subjects, without Fibrodysplasia Ossificans Progressiva (FOP).

22. The kit of claim 19, further comprising at least one standard, obtained from a subject, or pool of subjects, with Fibrodysplasia Ossificans Progressiva (FOP).