Gene therapy for treating Wilson's disease
Compositions and regimens useful in treating Wilson's Disease are provided. The compositions include recombinant adeno-associated virus (rAAV) with a transthyretin enhancer and promoter driving expression of a human ATP7B.
1. A recombinant adeno-associated virus (rAAV) useful as a liver-directed therapeutic for Wilson's Disease (WD), said rAAV comprising an AAV capsid, and a vector genome packaged therein, said vector genome comprising:
(a) an AAV 5′ inverted terminal repeat (ITR) sequence;
(b) a transthyretin (TTR) promoter sequence and a TTR enhancer sequence;
(c) a coding sequence encoding a truncated human copper-transporting ATPase 2 (ATP7B) comprising metal-binding domains (MBD) 4-6 and comprising a deletion of MBD_1-3; and
(d) an AAV 3′ ITR sequence.
2. The rAAV according to claim 1 , wherein the coding sequence of (c) is a native coding sequence.
3. The rAAV according to claim 1 , wherein the AAV capsid is an AAV8 capsid or variant thereof.
4. The rAAV according to claim 1 , wherein the TTR promoter sequence is a modified TTR promoter sequence.
5. The rAAV according to claim 1 , wherein the AAV 5′ ITR and/or AAV 3′ ITR sequence is from AAV2.
6. The rAAV according to claim 1 , wherein the vector genome further comprises a polyA signal sequence.
7. The rAAV according to claim 6 , wherein the polyA is about 75 bp in length.
8. The rAAV according to claim 1 , wherein the vector genome further comprises an intron sequence.
9. The rAAV according to claim 8 , wherein the intron sequence is from human beta globin IVS2 or SV40.
10. The rAAV according to claim 1 , wherein the vector genome is about 3 kilobases to about 5.5 kilobases in size.
11. A method of treating a patient having Wilson's Disease with the rAAV according to claim 1 , wherein the rAAV is delivered at about 1×10 12 to about 1×10 14 genome copies (GC)/kg in an aqueous suspension, wherein the GC are calculated as determined based on oqPCR or ddPCR.
12. The rAAV according to claim 1 , wherein the coding sequence of (c) is a codon-optimized sequence.
13. An aqueous suspension for administration to a Wilson's Disease patient, said suspension comprising an aqueous suspending liquid and about 1×10 12 GC/mL to about 1×10 14 GC/mL of a recombinant adeno-associated virus (rAAV) useful as a liver-directed therapeutic for Wilson's Disease, said rAAV having an AAV capsid, and having packaged therein a vector genome comprising:
(a) an AAV 5′ inverted terminal repeat (ITR) sequence;
(b) a TTR promoter sequence and a TTR enhancer sequence;
(c) a coding sequence encoding a truncated human copper-transporting ATPase 2 (ATP7B) comprising metal-binding domains (MBD) 4-6 and comprising a deletion of MBD_1-3; and
(d) an AAV 3′ ITR sequence.
14. The aqueous suspension according to claim 13 , wherein the suspension is suitable for intravenous injection.
15. The aqueous suspension according to claim 13 , wherein the suspension further comprises a surfactant, preservative, and/or buffer dissolved in the aqueous suspending liquid.
16. The aqueous suspension according to claim 13 , wherein the AAV capsid is an AAV8 capsid.