AAV-NAGLU vectors for treatment of mucopolysaccharidosis IIIB
View Patent ↗Recombinant nucleic acid comprising nucleotides encoding α-N-acetylglucosaminidase (NAGLU). Vectors comprising the recombinant nucleic acid for delivery to a subject. In an aspect the NAGLU sequences optimized for expression in human cells. Methods of using the vector to increase secretion of NAGLU from a cell and for treatment and prevention of mucopolysaccharidosis IIIB.
1 . A recombinant nucleic acid comprising a nucleotide sequence encoding human α-N-acetylglucosaminidase (NAGLU) that is codon-optimized for expression in human cells, wherein the recombinant nucleic acid comprises the nucleotide sequence of SEQ ID NO: 1.
2 . A vector comprising the recombinant nucleic acid of claim 1 .
3 . The vector of claim 2 , comprising the sequence of SEQ ID NO:3 or SEQ ID NO:4 or a sequence at least 90% identical thereto.
4 . An adeno-associated virus (AAV) vector genome comprising the recombinant nucleic acid of claim 1 .
5 . The AAV vector genome of claim 4 , wherein the AAV vector genome is a self-complementary AAV vector genome.
6 . The AAV vector genome of claim 4 , wherein the recombinant nucleic acid is operably linked to a constitutive promoter.
7 . The AAV vector genome of claim 6 , wherein the constitutive promoter is the CBA promoter.
8 . The AAV vector genome of claim 6 , wherein the constitutive promoter is the CMV promoter.
9 . A cell in vitro comprising the AAV vector genome of claim 4 .
10 . An AAV particle comprising the AAV vector genome of claim 4 .
11 . A method of producing a recombinant AAV particle comprising an AAV capsid, the method comprising:
providing a cell in vitro with AAV Cap and AAV Rep coding sequences, the AAV vector genome of claim 4 , and helper functions for generating a productive AAV infection; and
allowing assembly of the recombinant AAV particle comprising the AAV capsid and encapsidating the AAV vector genome.
12 . An AAV particle produced by the method of claim 11 .
13 . The AAV particle of claim 10 , wherein the AAV particle is an AAV9 particle.
14 . A pharmaceutical formulation comprising the AAV particle of claim 10 and a pharmaceutically acceptable carrier.
15 . A method of expressing NAGLU in a cell, comprising contacting the cell with an effective amount of the AAV particle of claim 10 , thereby expressing NAGLU in the cell.
16 . A method of increasing secretion of NAGLU from a cell, comprising contacting the cell with an effective amount of the AAV particle of claim 10 , thereby increasing secretion of NAGLU from the cell relative to the secretion of NAGLU after contacting the cell with an AAV particle comprising a nucleic acid comprising the wild-type sequence for NAGLU.
17 . A method of delivering NAGLU to a subject, comprising administering to the subject an effective amount of the pharmaceutical formulation of claim 14 , thereby delivering NAGLU to the subject.
18 . A method of treating or delaying the onset of mucopolysaccharidosis IIIB (MPS IIIB) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the pharmaceutical formulation of claim 14 , thereby treating or delaying the onset of MPS IIIB in the subject.
19 . The method of claim 18 , wherein the AAV particle is administered to the subject systemically.