Recombinant adeno-associated virus delivery of exon 2-targeted U7SNRNA polynucleotide constructs

• US 9,862,945 B2
• Filed: 04/18/2014
• Issued: 01/09/2018
• Est. Priority Date: 04/20/2013
• Status: Active Grant

• Current Assignee: Research Institute at Nationwide Children's Hospital
• Original Assignee: Research Institute at Nationwide Children's Hospital
• Inventors: Flanigan, Kevin, Vulin-Chaffiol, Adeline, Wein, Nicolas
• Primary Examiner(s): Nguyen, Quang
• Application Number:
  US14/785,769
• Publication Number:
  US 20160076028A1
• Time in Patent Office: 1,362 Days
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/0306 Animal model for genetic diseases
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: C07K 14/4708 Duchenne dystrophy
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• 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 15/86 Viral vectors
• CPC Class Codes: C12N 15/8645 Adeno-associated virus
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12N 2320/33 Alteration of splicing
• CPC Class Codes: C12N 2330/51 Specially adapted vectors
• CPC Class Codes: C12N 2750/14121 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2750/14132 Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2750/14171 Demonstrated in vivo effect
• CPC Class Codes: C12N 2750/14321 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2750/14333 : Use of viral protein as therapeutic agent other than vaccine, e.g. apoptosis inducing or anti-inflammatory
• CPC Class Codes: C12N 7/00 : Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

The present invention relates to recombinant adeno-associated virus (rAAV) delivery of polynucleotides for treating Duchenne Muscular Dystrophy resulting from the duplication of DMD exon 2. The invention provides rAAV products and methods of using the rAAV in the treatment of Duchenne Muscular Dystrophy.

Claims

1. A method of ameliorating Duchenne Muscular Dystrophy in a patient with DMD exon 2 duplications in need thereof comprising the step of administering a recombinant adeno-associated virus (rAAV) to the patient, wherein the recombinant AAV comprises the genome insert set out in SEQ ID NO:

2.

6. The method of any of claims 1-5 wherein the virus genome is a self-complementary genome.

8. The method of claim 1, 2, 3, 4, 5 or 7 wherein genome of the rAAV lacks AAV rep and cap DNA.

10. The method of claim 1, 2, 3, 4, 5 or 7 wherein the recombinant adeno-associated virus is a recombinant AAV rh74 virus, a recombinant AAV6 virus or a recombinant AAV9 virus.

2. A method of inhibiting the progression of dystrophic pathology associated with Duchenne Muscular Dystrophy in a patient with DMD exon 2 duplications in need thereof comprising the step of administering a rAAV to the patient, wherein the recombinant AAV comprises the genome insert set out in SEQ ID NO:

2.

3. A method of improving muscle function in a patient afflicted with Duchenne Muscular Dystrophy associated with DMD exon 2 duplications comprising the step of administering a rAAV to the patient, wherein the recombinant AAV comprises the genome insert set out in SEQ ID NO:

2.

4. The method of claim 3 wherein the improvement in muscle function is an improvement in muscle strength.

5. The method of claim 3 wherein the improvement in muscle function is an improvement in stability in standing and walking.

7. A method of delivering an exon 2-targeted U7snRNA polynucleotide construct to an patient with DMD exon 2 duplications, comprising the step of administering a rAAV to the patient, wherein the recombinant AAV comprises the genome insert set out in SEQ ID NO:

2.

9. The method of claim 7 wherein the virus genome is a self-complementary genome.

11. A recombinant adeno-associated virus (AAV), wherein the recombinant AAV comprises the genome insert set out in SEQ ID NO:

2.

13. The rAAV of claim 11 or 12 wherein genome of the rAAV lacks AAV rep and cap DNA.

14. The rAAV of claim 11 or 12 wherein the genome is a self-complementary genome.

12. A recombinant adeno-associated virus (AAV) comprising an AAV rh.74 capsid, an AAV6 capsid or an AAV9 capsid;

and the genome insert set out in SEQ ID NO;

2.