ALLELE-SPECIFIC SILENCING THERAPY FOR DYNAMIN 2-RELATED DISEASES

• US 20190309305A1
• Filed: 11/19/2017
• Published: 10/10/2019
• Est. Priority Date: 11/29/2016
• Status: Active Grant

• Current Assignee: Centre National De La Recherche Scientifique, Inserm (institut national de la sant et de la recherche mdicale, Institut de Myologie, Sorbonne Universite
• Original Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH), Association Isntitut de Myologie, Centre National De La Recherche Scientifique, Sorbonne Universite
• Inventors: BITOUN, Marc, TROCHET, Delphine, PRUDHON, Bernard
• Granted Patent: US 10,947,540 B2
• Time in Patent Office: Days
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2320/34 Allele or polymorphism specific uses
• CPC Class Codes: C12Y 306/05005 Dynamin GTPase (3.6.5.5)

Abstract

The invention relates to an allele specific siRNA able to silence the expression of only one allele of a heterozygous DNM2 gene, for treating diseases caused by heterozygous mutation and/or overexpression of Dynamin 2.

Claims

1. An allele specific siRNA (AS-siRNA) that comprises a sequence mismatch and is able to silence the expression of only one allele of a heterozygous DNM2 gene in a cell.

2. The AS-siRNA of claim 1, wherein the AS-siRNA is of 19-23 base pairs in length.

3. The AS-siRNA of claim 1 wherein the DNM2 gene is heterozygous for the presence of a non-pathological polymorphism, and/or is heterozygous for the presence of a disease-causing mutation.

4. The AS-siRNA of claim 3, wherein said AS-siRNA targets a region of a DNM2 gene transcript comprising said non-pathological polymorphism.

5. The AS-siRNA of claim 4, wherein said non-pathological polymorphism is on the same allele as the disease-causing mutation.

6. The AS-siRNA of claim 3, wherein the non-pathological polymorphism is rs2229920 (C or T) or rs12461992 (A or T).

7. AS-siRNA of claim 6, wherein the AS-siRNA is of 19 base pairs in length and wherein the position of the sequence mismatch is located at position N16 or N17 from the 5′

end of the sense strand of said AS-siRNA.

8. The AS-siRNA of claim 6, wherein the AS-siRNA comprises a sense strand selected from the group consisting of:

SEQ ID NO;

1, SEQ ID NO;

2, SEQ ID NO;

3 and SEQ ID NO;

4.

9. The AS-siRNA of claim 3, wherein said AS-siRNA targets a region of a DNM2 gene transcript comprising said disease-causing mutation.

10. The AS-siRNA of claim 9, wherein the disease-causing mutation is 1393C>

T;

c.1105C>

T, c.1106G>

A, c.1856C>

T or c.1948G>

A.

11. The AS-siRNA of claim 10, wherein the AS-siRNA is of 19 base pairs in length and wherein the position of the sequence mismatch is located at position N9, N10, N11, N12, NIS or N16 from the 5′

end of the sense strand of said AS-siRNA.

12. The AS-siRNA of claim 9, wherein the AS-siRNA comprises a sense strand selected from the group consisting of:

SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

8, SEQ ID NO;

9 and SEQ ID NO;

10.

13. The AS-siRNA of claim 1, wherein the AS-siRNA reduces expression of DNM2 mRNA and/or DNM2 protein by 20-60%.

14. A vector encoding the AS-siRNA according to claim 1.

15. A target cell, which is transfected or transduced with the vector according to claim 14.

16. An in vitro method for silencing the expression of a mutated allele of a DNM2 gene without silencing the expression of the wild type allele of the DNM2 gene in a target cell, comprising introducing in said target cell an AS-siRNA according to claim 1 or a vector encoding the AS-siRNA.

17. A method for treating a disease associated with an overexpression of Dynamin 2 in a subject in need thereof, comprising

administering to the subject a therapeutic amount of the AS-siRNA of claim 1, or a vector encoding the AS-siRNA, or a cell transfected or transduced with the vector.

18. A method for treating a disease induced by one or more disease-causing mutations in the DNM2 gene, in a subject in need thereof, comprising

administering to the subject a therapeutic amount of the AS-siRNA of claim 1, or a vector encoding the AS-siRNA, or a cell transfected or transduced with the vector.

19. The method of claim 17, wherein the disease associated with an overexpression of Dynamin 2 is X-linked myotubular myopathy, prostate cancer or pancreatic cancer.

20. The method of claim 18, wherein the disease induced by the one or more disease-causing mutations in the DNM2 gene is autosomal dominant centronuclear myopathy, T-cell acute lymphoblastic leukemia, Charcot-Marie-Tooth disease or Hereditary Spastic Paraplegia.