Gene therapy for mucopolysaccharidosis iiib

Abstract

Provided herein is a recombinant AAV (rAAV) comprising an AAV capsid and a vector genome packaged therein, wherein the vector genome comprises an AAV 5′ inverted terminal repeat (ITR), an engineered nucleic acid sequence encoding a functional human N-acetyl-alpha-glucosaminidase (hNAGLU), a regulatory sequence which direct expression of hNAGLU in a target cell, and an AAV 3′ ITR. Also provided is a pharmaceutical composition comprising a rAAV as described herein in a formulation buffer, and a method of treating a human subject diagnosed with MPS IIIB.

1 . A pharmaceutical composition comprising a plurality of recombinant AAV (rAAV) comprising an AAV capsid and a vector genome packaged therein, wherein the vector genome comprises an AAV 5′ inverted terminal repeat (ITR), an engineered nucleic acid sequence encoding a functional human N-acetyl-alpha-glucosaminidase (hNAGLU), a regulatory sequence which directs expression of hNAGLU in a target cell, and an AAV 3′ ITR, wherein the hNAGLU coding sequence is at least 95% identical to SEQ ID NO: 1. 
     
     
         2 . The pharmaceutical composition according to  claim 1 , wherein the hNAGLU coding sequence is SEQ ID NO:1. 
     
     
         3 . The pharmaceutical composition according to  claim 1 , wherein the AAV vector genome comprises the sequence of SEQ ID NO: 4. 
     
     
         4 . The pharmaceutical composition according to  claim 1 , wherein the AAV capsid is an AAV9 capsid. 
     
     
         5 . The pharmaceutical composition according to  claim 1  further comprising a formulation buffer. 
     
     
         6 . The pharmaceutical composition according to  claim 5 , which is formulated for delivery via intracerebroventricular (ICV), intrathecal (IT), intracisternal or intravenous (IV) injection. 
     
     
         7 . A method of treating a human subject diagnosed with MPS IIIB and/or improving gait or mobility, reducing tremors, reducing spasms, improving posture, or reducing the progression of vision loss in a subject in need thereof, comprising administering to the subject a composition comprising the rAAV in a dose of 1×10 9  GC per gram of brain mass to about 1×10 13  GC per gram of brain mass. 
     
     
         8 . The method according to  claim 7 , wherein said method results in a serum NAGLU activity at least about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% of a healthy control. 
     
     
         9 . The method according to  claim 7 , wherein the suspension is suitable for co-administering with a functional hNAGLU protein. 
     
     
         10 . The method according to  claim 7 , wherein the suspension is delivered into the subject in need intracerebroventricularly, intrathecally, or intravenously. 
     
     
         11 . A nucleic acid molecule comprising an engineered nucleic acid sequence encoding a functional hNAGLU and a regulatory sequence which directs expression thereof in a target cell, wherein the hNAGLU coding sequence is SEQ ID NO: 1 or at least 95% identical to SEQ ID NO: 1. 
     
     
         12 . The nucleic acid molecule according to  claim 11 , wherein the hNAGLU coding sequence is SEQ ID NO: 1. 
     
     
         13 . The nucleic acid molecule according to  claim 11 , which is a plasmid. 
     
     
         14 . A cell comprising the nucleic acid molecule according to  claim 11 . 
     
     
         15 . The cell according to  claim 14 , wherein the cell is a yeast cell, a human cell, a non-human cell, a mammalian cell, a non-mammalian cell, or an insect cell. 
     
     
         16 . The cell according to  claim 14  which is an HEK-293 cell.