Compositions and methods for treating retinal disorders
The present invention relates to the prevention and/or treatment of retinal disorders, such as cone dystrophies, cone-rod dystrophies, in particular Achromatopsia.
1. A transcriptional control unit (TCU) of up to 2500 nucleotides in length comprising in a 5′ to 3′ direction:
(a) a Locus Control Region (LCR) comprising
(i) SEQ ID NO: 1; or
(ii) a sequence having at least 90% sequence identity to said sequence (a)(i); and
(b) a promoter element comprising SEQ ID NO: 16 and
(i) at least the last 200 nucleotides but no more than 1100 nucleotides of SEQ ID NO: 17; or
(ii) a sequence having at least 90% sequence identity to said sequence (b)(i).
2. The TCU of claim 1 , wherein (b) comprises:
(i) at least the last 500 nucleotides SEQ ID NO: 17, or
(ii) a sequence having at least 90% sequence identity to said sequence (i).
3. The TCU of claim 1 , wherein (b) comprises at least 200 nucleotides of SEQ ID NO: 3.
4. The TCU of claim 1 , wherein the sequence of (b)(i) comprises a sequence of at least 10 contiguous nucleotides selected from nucleotides 1442 to 1476 of SEQ ID NO:17, or wherein the sequence of (b)(ii) comprises a sequence comprising at least 10 contiguous nucleotides selected from a sequence having at least 90% sequence identity to nucleotides 1442 to 1476 of SEQ ID NO:17.
5. The TCU according to claim 1 , wherein (b) comprises:
(i) SEQ ID NO: 3; or
(ii) a sequence having at least 90% sequence identity to said sequence (i).
6. The TCU of claim 5 , wherein the TCU comprises SEQ ID NO: 4 or SEQ ID NO: 15.
7. The TCU of claim 1 , wherein the promoter element comprises SEQ ID NO: 5.
8. The TCU of claim 7 , wherein the TCU comprises SEQ ID NO: 6.
9. An expression construct comprising the TCU of claim 1 , wherein the TCU is operably linked to a gene sequence to be expressed.
10. The expression construct according to claim 9 , wherein the operably linked gene sequence is CNGA3, CNGB3, PDE6C, PDE6H, GNAT2, KCNV2 or CACNA2D4.
11. The expression construct according to claim 10 , wherein the operably linked gene sequence comprises SEQ ID NO: 7, 8, 9, 10, 11, 12, 13 or 14, or that has at least 80% sequence identity to SEQ ID NO: 7, 8, 9, 10, 11, 12, 13 or 14 and has the ability to rescue cone photoreceptor function.
12. The expression construct according to claim 10 , wherein the operably linked gene sequence comprises SEQ ID NO: 8, or that has at least 80% sequence identity to SEQ ID NO: 8 and has the ability to rescue cone photoreceptor function.
13. A vector comprising the TCU according to claim 1 .
14. The vector according to claim 13 , wherein the vector is a viral vector.
15. The vector according to claim 14 , wherein the vector is an adeno-associated virus (AAV) vector.
16. The vector according to claim 15 , wherein the vector comprises an AAV genome or a derivative thereof.
17. The vector according to claim 16 , wherein the AAV comprises a capsid that is derived from AAV8.
18. The vector according to claim 17 , wherein the TCU is operably linked to a gene sequence to be expressed.
19. The vector according to claim 18 , wherein the operably linked gene sequence is CNGA3.
20. The vector according to claim 16 , wherein said derivative is a chimeric, shuffled or capsid modified derivative.
21. The vector according to claim 16 , wherein said AAV genome is from a naturally derived serotype or isolate or clade of AAV.
22. The vector according to claim 21 , wherein said AAV genome is from AAV serotype 2 (AAV2), AAV serotype 4 (AAV4), or AAV serotype 8 (AAV8).
23. The vector according to claim 22 , wherein the genome is derived from AAV2.
24. An isolated host cell that contains the vector of claim 13 .
25. The isolated host cell according to claim 24 that is a HEK293 or HEK293T cell.
26. A pharmaceutical composition comprising the vector of claim 13 and a pharmaceutically acceptable carrier.
27. A method of treating a retinal disorder in a patient in need thereof, comprising administering a therapeutically effective amount of the vector according to claim 13 to said patient.
28. The method of claim 27 , wherein the retinal disorder is Achromatopsia.
29. The method of claim 27 , wherein the treatment is by administration of the vector to a patient by direct retinal, subretinal or intravitreal injection.
30. The method according to claim 27 , wherein said vector is administered directly into the retinal, subretinal space or intravitreal space.
31. A vector comprising the expression construct according to claim 9 .
32. The vector according to claim 31 , wherein the vector is a viral vector.
33. The vector according to claim 32 , wherein the vector is an AAV vector.
34. The vector according to claim 33 , wherein the vector comprises an AAV genome or a derivative thereof.
35. The vector according to claim 34 , wherein the AAV comprises a capsid that is derived from AAV8.
36. The vector according to claim 35 , wherein said AAV genome is from AAV serotype 2 (AAV2), AAV serotype 4 (AAV4), or AAV serotype 8 (AAV8).
37. The vector according to claim 36 , wherein the genome is derived from AAV2.
38. The vector according to claim 37 , wherein the operably linked gene sequence is CNGA3.
39. An isolated host cell that produces the viral vector of claim 13 .
40. The isolated host cell according to claim 39 , wherein the cell is a HEK293 or HEK293T cell.
41. The expression construct of claim 9 , wherein the TCU comprises SEQ ID NO: 4 and wherein the TCU is operably linked to SEQ ID NO: 8.
42. The expression construct according to claim 9 , wherein the operably linked gene sequence is CNGA3 and the TCU comprises SEQ ID NO: 15.