THERAPEUTIC INTERVENTION IN A GENETIC DISEASE IN AN INDIVIDUAL BY MODIFYING EXPRESSION OF AN ABERRANTLY OR ABNORMALLY EXPRESSED GENE

• US 20100125099A1
• Filed: 04/20/2007
• Published: 05/20/2010
• Est. Priority Date: 04/20/2006
• Status: Active Grant

• Current Assignee: Academisch Ziekenhuis Leiden
• Original Assignee: Academisch Ziekenhuis Leiden
• Inventors: van Ommen, Garrit Jan Boudewijn, Sterrenburg, Petronella Johanna Elisabeth, den Dunnen, Johannes Theodorus, 't Hoen, Peter Abraham Christiaan
• Granted Patent: US 8,304,398 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61K 38/185 Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/33 Alteration of splicing
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2502/1335 Skeletal muscle cells, myocytes, myoblasts, myotubes
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0658 Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts

Abstract

The present invention provides means and methods for alleviating genetic disease. A genetic defect that has a phenotype in differentiated cells can lead to defects in precursor cells thereof. These so-called secondary defects contribute to the overall disease of the individual. In the present invention, genetic intervention with the aim to alleviate symptoms of genetic disease is directed toward the primary genetic defect in the differentiated cell and the secondary defect in the precursor cell.

Claims

1. Use of a compound for reducing, inhibiting and/or antagonizing expression of Bone Morphogenetic Protein 4 (BMP4) in a cell, for the manufacture of a medicament for alleviating a symptom of a genetic muscular dystrophy in an individual.

2. Use according to claim 1, wherein said cell is a myoblast cell or a precursor thereof.

3. Use according to claim 1, wherein said compound comprises an antisense RNA or a functional equivalent thereof.

11. Use according to claim 1, wherein expression is decreased, inhibited and/or antagonized by means of a virally transduced DNA sequence.

12. Use according to claim 1, wherein said compound is provided to said cell by means of a viral vector.

13. Use or method according to claim 1, wherein said genetic muscular dystrophy comprises one of the following diseases:

Duchenne Muscular Dystrophy (DMD), Becker Muscular Dystrophy (BMD), Congenital Muscular Dystrophy (CMD), Distal Muscular Dystrophy (DD), Emery-Dreifuss Muscular Dystrophy (EDMD), Facioscapulohumeral Muscular Dystrophy (FSH), Limb-Girdle Muscular Dystrophy (LGMD), Myotonic Dystrophy (MMD), Oculopharyngeal Muscular Dystrophy (OPMD).

14. Use or method according to claim 1, wherein said compound comprises a protein inhibitor.

15. Use or method according claim 1, wherein said compound comprises a BMP4 inhibitor or antagonist.

16. Use or method according to claim 14, wherein said inhibitor or antagonist comprises noggin, chordin, ventroptin, twisted gastrulation, gremlin or other members of the DAN family of BMP4 antagonists, PRDC, sclerostin, CTGF and/or follistatin or a functional part, derivative and/or analogue thereof.

17. Use or method according to claim 1, further comprising use of a second compound for the preparation of a medicament for treatment of said individual wherein said second compound provides a muscle cell of said individual with at least part of the normal function of a gene that is associated with said genetic muscular dystrophy (mutated gene).

18. Use or method according to claim 1, wherein said genetic muscular dystrophy is Duchenne Muscular Dystrophy (DMD) or Becker Muscular Dystrophy (BMD).

19. Use according to claim 18, wherein said second compound comprises an oligonucleotide, or functional equivalent thereof, for skipping an exon of a dystrophin gene.

4. A method for stimulating differentiation of a myoblast cell comprising providing said myoblast cell with a compound for reducing, inhibiting and/or antagonizing expression of BMP4 in said myoblast cell.

5. A method according to claim 4 wherein said compound decreases and/or inhibits BMP4 mRNA expression in said myoblast cell.

6. A method according to claim 4, further comprising contacting said myoblast cell with a mature muscle cell.

7. A method according to claim 4, wherein said mature muscle cell is derived from a subject that suffers from said genetic muscular dystrophy.

8. A method according to claim 4, wherein reducing and/or inhibiting BMP4 mRNA expression in said myoblast cell comprises providing said myoblast cell or a precursor thereof with a BMP4 oligonucleotide that is complementary to said BMP4 gene (anti-sense).

9. A method according to claim 4, wherein said myoblast cells or precursors thereof are in vitro provided with said BMP-4 anti-sense oligonucleotide.

29. A method according to claim 4, wherein expression is decreased, inhibited and/or antagonized by means of a virally transduced DNA sequence.

30. A method according to claim 4, wherein said compound is provided to said cell by means of a viral vector.

10. A collection of myoblast cells or precursors thereof comprising a BMP-4 anti-sense oligonucleotide.

20. A method for determining whether a BMP4 anti-sense oligonucleotide or a functional equivalent thereof is capable of inducing skipping of an exon in a BMP4 pre-mRNA containing said exon, said method comprising providing a cell expression said BMP4 pre-mRNA with said oligonucleotide and determining whether said exon is absent from mature mRNA produced from said pre-mRNA.

21. A method according to claim 20, wherein said anti-sense oligonucleotide or a functional equivalent thereof is complementary to said exon.

22. A method according to claim 20, wherein said anti-sense oligonucleotide or a functional equivalent thereof is complementary to an exon-internal part of said exon.

23. A method according to claim 20, wherein said anti-sense oligonucleotide or a functional equivalent thereof is complementary to exon 4 of BMP4.

27. Use or a method according to claim 20, wherein said anti-sense oligonucleotide is provided to said cell by means of a viral vector.

28. Use according to claim 27, wherein said viral vector comprises an expression cassette for expression of said anti-sense oligonucleotide.

24. Use of an anti-sense oligonucleotide or a functional equivalent thereof that is complementary to an exon of BMP-4, for skipping said exon in a BMP4 pre-mRNA.

25. An anti-sense oligonucleotide or a functional equivalent thereof is complementary to an exon of BMP-4 for use in the treatment of a genetic muscular dystrophy.

26. Use of an anti-sense oligonucleotide or a functional equivalent thereof is complementary to an exon of BMP-4 for the preparation of a medicament for the treatment of a genetic muscular dystrophy.