Microdystrophin gene therapy constructs and uses thereof
AssigneeREGENXBIO Inc.
InventorsChunping Qiao, Devin McDougald, Ye Liu, Olivier Danos
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Abstract
Provided are gene constructs that encode a microdystrophin protein for use in gene therapy. The microdystrophin gene constructs and expression cassettes were engineered for improved therapy with respect to efficacy, potency and safety to the subject when expressed by a viral vector in muscle cells and/or CNS cells.
Claims51
A recombinant adeno-associated virus (rAAV) particle comprising a) a nucleic acid comprising a nucleotide sequence encoding a microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 79, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein.
A method of delivering a nucleic acid encoding a microdystrophin protein to a cell, the method comprising contacting the cell with an rAAV particle, wherein the rAAV particle comprises a) a nucleic acid comprising a nucleotide sequence encoding the microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 79, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein, wherein the nucleic acid encoding the microdystrophin protein is delivered to the cell.
An rAAV particle comprising a) a nucleic acid comprising a nucleotide sequence encoding a microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 1, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein.
An rAAV particle comprising a) a nucleic acid comprising 5′ to 3′:
- a first AAV ITR sequence-a transcription regulatory element sequence-a nucleotide sequence encoding a microdystrophin protein-a polyadenylation sequence-a second AAV ITR sequence,
- wherein the microdystrophin protein comprises the amino acid sequence of SEQ ID NO:
- 1, wherein the transcription regulatory element is an SPc5-12 promoter or a transcriptionally active portion thereof, and
- b) a capsid comprising a capsid protein comprising the amino acid sequence of SEQ ID NO:
- 77.
A pharmaceutical composition comprising i) a rAAV particle comprising a) a nucleic acid comprising a nucleotide sequence encoding a microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 79, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein, and ii) a pharmaceutically acceptable carrier.
A pharmaceutical composition comprising i) a rAAV particle comprising a) a nucleic acid comprising a nucleotide sequence encoding a microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 1, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein, and ii) a pharmaceutically acceptable carrier.
A pharmaceutical composition comprising i) a rAAV particle comprising a) a nucleic acid comprising 5′ to 3′:
- a first AAV ITR sequence-a transcription regulatory element sequence-a nucleotide sequence encoding a microdystrophin protein-a polyadenylation sequence-a second AAV ITR sequence,
- wherein the microdystrophin protein comprises the amino acid sequence of SEQ ID NO:
- 1,
- wherein the transcription regulatory element is an SPc5-12 promoter or a transcriptionally active portion thereof, and b) a capsid comprising a capsid protein comprising the amino acid sequence of SEQ ID NO:
- 77, and ii) a pharmaceutically acceptable carrier.
A method of delivering a nucleic acid encoding a microdystrophin protein to a cell, the method comprising contacting the cell with an rAAV particle, wherein the rAAV particle comprises a) the nucleic acid comprising a nucleotide sequence encoding the microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 1, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein, wherein the nucleic acid encoding the microdystrophin protein is delivered to the cell.
A method of treating a dystrophinopathy in a human subject in need thereof, the method comprising:
- administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a rAAV particle, wherein the rAAV particle comprises a) a nucleic acid comprising a nucleotide sequence encoding a microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 79, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein,
- wherein the administration results in expression of the microdystrophin protein in a muscle cell of the subject thereby treating the dystrophinopathy in the subject.
A method of treating a dystrophinopathy in a human subject in need thereof, the method comprising:
- administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a recombinant adeno-associated virus (rAAV) particle, wherein the rAAV particle comprises a) a nucleic acid comprising a nucleotide sequence encoding a microdystrophin protein comprising the amino acid sequence of SEQ ID NO:
- 1, or the reverse complement of the nucleotide sequence, and b) a capsid comprising a capsid protein,
- wherein the administration results in expression of a microdystrophin protein in a muscle cell of the subject thereby treating the dystrophinopathy in the subject.
A method of treating Duchenne muscular dystrophy (DMD) in a human subject in need thereof, the method comprising:
- administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a recombinant adeno-associated virus (rAAV) particle, wherein the rAAV particle comprises
- a) a nucleic acid comprising from 5′ to 3′:
- a first AAV ITR sequence-a transcription regulatory element sequence-a nucleotide sequence encoding a microdystrophin protein-a polyadenylation sequence-a second AAV ITR sequence,
- wherein the microdystrophin protein comprises the amino acid sequence of SEQ ID NO:
- 1,
- wherein the transcription regulatory element is an SPc5-12 promoter or a transcriptionally active portion thereof, and
- b) a capsid comprising a capsid protein comprising the amino acid sequence of SEQ ID NO:
- 77, wherein the administration results in expression of the microdystrophin protein in a muscle cell of the subject thereby treating the DMD in the subject.
Dependencies are inferred from the claim text.