Compositions and methods for the treatment of disorders related to glucosylceramidase beta 1 deficiency

The AAV capsid variant enhances GBA1 protein levels to treat or prevent GBA1-related disorders like Parkinson's disease and Gaucher disease by delivering the protein effectively.

HK40134653APending Publication Date: 2026-07-10VOYAGER THERAPEUTICS INC
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Patent Information

Authority / Receiving Office
HK · HK
Patent Type
Applications
Current Assignee / Owner
VOYAGER THERAPEUTICS INC
Filing Date
2026-04-07
Publication Date
2026-07-10

AI Technical Summary

Technical Problem

Current treatments for GBA1-related disorders such as Parkinson's disease, Gaucher disease, Parkinson's dementia, and Lewy body dementia do not effectively address the underlying deficiency of the GBA1 protein, leading to inadequate therapeutic outcomes.

Method used

Utilizing an adeno-associated virus (AAV) capsid variant to deliver and enhance the level of the GBA1 protein, thereby treating or preventing these disorders.

Benefits of technology

The AAV capsid variant effectively increases GBA1 protein levels, providing therapeutic benefits for subjects with or at risk of GBA1-related disorders.

✦ Generated by Eureka AI based on patent content.

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Abstract

The disclosure relates to compositions and methods for, inter alia, altering, e.g., enhancing, the level of GBA1 protein via delivery using an adeno-associated viral (AAV) capsid variant. The compositions and methods of the present disclosure are useful, inter alia, in the treatment of subjects who have, have been diagnosed with, or are at risk of having a GBA1-related disorder, e.g., Parkinson's Disease (PD), Gaucher Disease (GD), Parkinson's Disease Dementia (PDD), Dementia with Lewy Bodies (DLB), or Lewy Body Dementia (LBD).
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Description

CN Title: Compositions and Methods for Treating Disorders Related to Glucosinolate β1 Deficiency CN Abstract: This disclosure relates to compositions and methods for delivering, particularly altering, for example enhancing, the level of the GBA1 protein, via the use of an adeno-associated virus (AAV) capsid variant. The compositions and methods disclosed herein are particularly useful for treating subjects who have already had, have been diagnosed with, or are at risk of having a GBA1-related disorder, such as Parkinson's disease (PD), Gaucher disease (GD), Parkinson's dementia (PDD), dementia with Lewy bodies (DLB), or Lewy body dementia (LBD). 1 Abstract

Claims

CLAIMSWhat is claimed is:1 . An adeno-associated virus (AAV) particle comprising: a) an AAV capsid variant comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3], wherein:(i) optionally [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G;(ii) [N2] comprises the amino acid sequence of SPH; and(iii) [N3] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid; and b) a viral genome comprising a p-glucocerebrosidase 1 (GBA1)-encoding sequence.

2. The AAV particle of claim 1, wherein the amino acid sequence [N1]-[N2]-[N3] is in hypervariable loop IV of the AAV capsid variant.The AAV particle of claim 1 or claim 2. wherein the AAV capsid variant is an AAV9 capsid variant.

4. The AAV particle of any one of claims 1-3, wherein [N 1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G.

5. The AAV particle of any one of claims 1-4, wherein [N2]-[N3] comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941).

6. An adeno-associated virus (AAV) particle comprising a viral genome comprising a β - glucocerebrosidase 1 (GBA1)-encoding sequence and an AAV9 capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941).

7. The AAV particle of claim 6. wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is in hypervariable loop IV of the AAV9 capsid variant.

8. The AAV particle of claim 6 or claim 7, wherein the amino acid sequence of SPHSKA ( SEQ ID NO: 941) is present immediately subsequent to an amino acid position corresponding to position 455 of SEQ ID NO: 4 or SEQ ID NO: 36.

9. The AAV particle of any one of claims 6-8, wherein the AAV9 capsid variant further comprises one, two, or all of: an N at an amino acid position corresponding to position 452, an E at an aminoacid position corresponding to position 451, and / or a V at an amino acid position corresponding to position 453 of SEQ ID NO: 4.

10. The AAV particle of claim any one of claims 6-9, wherein the AAV9 capsid variant comprises the amino acid sequence of KTENVSGSPHSKAQNQQT (SEQ ID NO: 3272).

11. The AAV particle of any one of claims 6-10, wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 4;(ii) a VP2 protein comprising an amino acid sequence having at least 90% identity to positions 138-742 of SEQ ID NO: 4; and / or(iii) a VP3 protein comprising an amino acid sequence having at least 90% identity to positions 203-742 of SEQ ID NO: 4.

12. The AAV particle of any one of claims 6-11. wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising an amino acid sequence having at least 95% identity to SEQ ID NO: 4;(ii) a VP2 protein comprising an amino acid sequence having at least 95% identity to positions 138-742 SEQ ID NO: 4; and / or(iii) a VP3 protein comprising an amino acid sequence having at least 95% identity to positions 203-742 of SEQ ID NO: 4.

13. The AAV particle of any one of claims 6-12, wherein the AAV9 capsid variant comprises :(i) a VP1 protein comprising an amino acid sequence having at least 99% identity to SEQ ID NO: 4;(ii) a VP2 protein comprising an amino acid sequence having at least 99% identity to positions 138-742 of SEQ ID NO: 4; and / or(iii) a VP3 protein comprising an amino acid sequence having at least 99% identity to positions 203-742 of SEQ ID NO: 4.

14. The AAV particle of any one of claims 6-13, wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:4.

15. The AAV particle of any one of claims 6-13. wherein the AAV9 capsid variant comprises:(i) the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to an amino acid position corresponding to position 455 of SEQ ID NO: 4:(ii) an E at an amino acid position corresponding to position 451 and a V at an amino acid position corresponding to position 453 of SEQ ID NO: 4; and(iii) no other modifications relative to wild type AAV9.

16. The AAV particle of any one of claims 6-8, wherein the AAV9 capsid variant further comprises one, two, or all of: an E at an amino acid position corresponding to position 451 , an R at an amino acid position corresponding to position 452, and / or a V at an amino acid position corresponding to position 453 of SEQ ID NO: 36.

17. The AAV particle of any one of claims 6-8 and 16, wherein the A.AV9 capsid variant comprises the amino acid sequence of KTERVSGSPHSK.AQNQQT (SEQ ID NO: 3589).

18. The AAV particle of any one of claims 6-8, 16, and 17, wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 36;(ii) a VP2 protein comprising an amino acid sequence having at least 90% identity to positions 138-742 SEQ ID NO: 36; and / or(iii) a VP3 protein comprising an amino acid sequence having at least 90% identity to positions 203-742 of SEQ ID NO: 36.

19. The AAV particle of any one of claims 6-8 and 16-18, wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising an amino acid sequence having at least 95% identity to SEQ ID NO: 36;(ii) a VP2 protein comprising an amino acid sequence liaving at least 95% identity to positions 138-742 SEQ ID NO: 36; and / or(iii) a VP3 protein comprising an amino acid sequence liaving at least 95% identity to positions 203-742 of SEQ ID NO: 36.

20. The AAV particle of any one of claims 6-8 and 16-19, wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising an amino acid sequence having at least 99% identity to SEQ IDNO: 36;(ii) a VP2 protein comprising an amino acid sequence having at least 99% identity to positions 138-742 of SEQ ID NO: 36; and / or(iii) a VP3 protein comprising an amino acid sequence having at least 99% identity to positions 203-742 of SEQ ID NO: 36.

21. The AAV particle of any one of claims 6-8 and 16-20, wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 36.

22. The AAV particle of any one of claims 6-8 and 16-20, wherein the AAV9 capsid variant comprises:(i) the amino acid sequence SPHSKA (SEQ ID NO: 941 ), wherein the amino acid sequence is present immediately subsequent to an amino acid position corresponding to position 455 of SEQ ID NO: 36;(ii) an E at an amino acid position corresponding to position 451, an R at an amino acid position corresponding to position 452, and a V at an amino acid position corresponding to position 453 of SEQ ID NO: 36; and(iii) no other modifications relative to wild type AAV9.

23. The AAV particle of any one of claims 1-4, wherein [N1]-[N2]-[N3] is present immediately subsequent to a position corresponding to the amino acid position 452 of SEQ ID NO: 982; and wherein the AAV capsid variant comprises an amino acid sequence at least 90% identical, e.g., at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% identical, to the amino acid sequence of SEQ ID NO: 982, e.g., to positions 203-742 of SEQ ID NO: 982,24. The AAV particle of claim 23, wherein [N1] comprises GHD.

25. The AAV particle of claim 23 or claim 24, wherein [N1] comprises the amino acid G at a position corresponding to position 453, the amino acid H at position 454, and the amino acid D at position 455 of SEQ ID NO: 138 or SEQ ID NO: 982.

26. The AAV particle of any one of claims 23-25, wherein [N3] comprises KSG.

27. The AAV particle of any one of claims 23-26, wherein the AAV capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO:

982. or an amino acid sequence having at least 90% identity to SEQ ID NO: 982;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO:982 or an amino acid sequence having at least 90% identity to positions 138-742 SEQ ID NO: 982; or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:982 or an amino acid sequence having at least 90% identity to positions 203-742 of SEQ ID NO: 982.

28. The AAV particle of any one of claims 23-27, wherein the AAV capsid variant comprises:(i) a ATI protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity to SEQ ID NO: 982;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity to positions 138-742 SEQ ID NO: 982; or(iii) a VP3 protein comprising the amino acid sequence of positions 2.03-742. of SEQ ID NO:

982. or an amino acid sequence having at least 95% identity to positions 203-742 of SEQ ID NO: 982.

29. The AAV particle of any one of claims 23-28, wherein the AAV capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity to SEQ ID NO: 982;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity to positions 138-742 SEQ ID NO: 982; or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity to positions 203-742 of SEQ ID NO: 982.

30. The AAV particle of any one of claims 23-29, wherein the AAV capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982;(ii) a AT2 protein comprising the amino acid sequence of positions 138-742. of SEQ ID NO: 982; or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982.

31. The AAV particle of any one of claims 1-30, wherein the viral genome encodes a wildtype GBA1 protein.

32. The AAV particle of any one of claims 1-31, wherein the viral genome does not encode a hemagglutinin (HA) tag.

33. The AAV particle of any one of claims 1-32, wherein the viral genome encodes a human GBA1 protein, a dog GBA1 protein, or an equine GBA1 protein.

34. The AAV particle of claim 33, wherein the viral genome encodes a human GBA1 protein, optionally wherein the human GBA1 protein comprises the amino acid sequence of SEQ ID NO: 1775.

35. The AAV particle of any one of claims 1-34, wherein the GBA1-encoding sequence comprises a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 2002.

36. The AAV particle of any one of claims 1 -35, wherein the GBA1-encoding sequence comprises a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 2002.

37. The AAV particle of any one of claims 1-36, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002.

38. The AAV particle of any one of claims 1-37, wherein the GBA1-encoding sequence encodes a signal sequence comprising an amino acid sequence that is at least 90% identical to SEQ ID: 2005.

39. The AAV particle of claim 38, wherein the signal sequence comprises the amino acid sequence of SEQ ID NO: 2005.

40. The AAV particle of any one of claims 1-39, wherein the GBA1-encoding sequence comprises a nucleotide sequence that is at least 90% identical (e.g,, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%. or 100% identical) to SEQ ID NO: 2001.

41. The AAV particle of claim 40, wherein the GBA1-encoding sequence comprises SEQ ID NO: 2001.

42. The AAV particle of any one of claims 1-41, wherein the viral genome further comprises a miRNA (miR) binding site that modulates expression of the encoded GBA1 protein in a cell or tissue of the DRG, liver, heart, hematopoietic lineage, or a combination thereof43. The AAV particle of any one of claims 1-42, wherein the viral genome further comprises a promoter operably linked to the GBA1 -encoding sequence.

44. The AAV particle of claim 43, wherein the promoter is at least 90% identical (e.g., at least 90%, at least 91% at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1834.

45. The AAV particle of claim 43, wherein the promoter comprises the nucleotide sequence of SEQ ID NO: 1834.

46. The AAV particle of any one of claims 1 -45, wherein the viral genome further comprises an enhancer.

47. The AAV particle of claim 46, wherein the enhancer compri ses a CMV immediate-early (CMVie) enhancer.

48. The AAV particle of claim 47, wherein the CMVie enhancer is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1831.

49. The AAV particle of claim 47, wherein the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831.

50. The AAV particle of any one of claims 1-49, wherein the viral genome further comprises an intron.51 . The AAV particle of claim 50, wherein the intron is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1842.

52. The AAV particle of claim 50, wherein the intron comprises the nucleotide sequence of SEQ ID NO: 1842.

53. The AAV particle of any one of claims 1-52, wherein the viral genome further comprises a polyadenylation (poly A) region.

54. The AAV particle of claim 53, wherein the poly A region is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1846.

55. The AAV particle of claim 53, wherein the poly A region comprises the nucleotide sequence of SEQ ID NO: 1846.

56. The AAV particle of any one of claims 1-55, wherein the viral genome further comprises an inverted terminal repeat (ITR).

57. The AAV particle of claim 56, wherein the ITR is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830.

58. The AAV particle of claim 56, wherein the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830.

59. The AAV particle of claim 56, wherein the viral genome comprises a 5’ ITR and a 3’ ITR, wherein the 5 ’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829 and the 3 ’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

60. The AAV particle of any one of claims 1-59, wherein the viral genome further comprises one or more miR183 binding sites.61 . The AAV particle of claim 60 wherein the viral genome comprises four miR 183 binding sites.

62. The AAV particle of claim 61, wherein each of the four miR183 binding sites comprises at least 70% identity to the nucleotide sequence of SEQ ID NO: 1847, optionally wherein each of the four miR183 binding sites comprises the nucleotide sequence of SEQ ID NO: 1847.

63. The AAV particle of any one of claims 1-60, wherein the viral genome further comprises a miR183 binding site series that comprises a sequence at least 95% (e.g., at least 95%, at least 96%, atleast 97%, at least 98%, at least 99%, or 100%) identical to the nucleotide sequence of SEQ ID NO: 1849.

64. The AAV particle of claim 63, wherein the miR183 binding site series comprises a nucleotide sequence of SEQ ID NO: 1849.

65. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeal (ITR);(ii) a promoter;(iii) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and(iv) a 3’ ITR.

66. The AAV particle of any one of claims 1 -34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5' inverted terminal repeat (ITR);(ii) a promoter;(iii) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence al least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001; and(iv) a 3’ ITR.

67. The AAV particle of any one of claims 1-34, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CB A) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, al least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1 -encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002; and(vi ) a 3 ’ ITR68. The AAV particle of any one of claims 1-34, wherein the viral genome comprises:(i) a 5’ inverted terminal repeal (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95% at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842; and(v) the GBA1-encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001 and(vi) a 3 ' ITR.

69. The AAV particle of any one of claims 1-34, wherein the viral genome comprises:(i) a 5 ’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVre) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002;(vi) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1846; and(vii) a 3’ H R.

70. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3 order:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001;(vi) a polyadenylation (polyA) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1846; and(vii) a 3’ ITR.

71. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002;(vi) a poly adenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and(vii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1830,72. The AAV particle of any one of claims 1-34, wherein the viral genome comprises:(i) a 5’ inverted terminal repeal (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g., al ieast 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at ieast 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at ieast 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence al least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001;(vi) a poly adenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at ieast 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and(vii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

73. The AAV particle of any one of claims 1-34, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2.006 or a nucleotide sequence at least 90% identical (e.g., at least 97% identical) to SEQ ID NO: 2006.

74. The AAV particle of claim 73, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006.

75. The AAV particle of claim 74, wherein the viral genome consists of the nucleotide sequence of SEQ ID NO: 2006.

76. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002;(iv) at least one miR 183 binding site; and(v) a 3’ ITR.

77. The AAV particle of any one of claims 1 -34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001;(iv) at least one miR183 binding site; and(v) a 3’ ITR.

78. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g.. at least 93% identical) to SEQ ID NO: 2.002;(iv) at least one miR 183 binding site series comprising at least one miR183 binding site and at least one spacer sequence; and(v) a 3’ ITR.

79. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat ( ITR);(ii) a promoter;(iii) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2.001 ;(iv) at least one miR183 binding site series comprising at least one miR183 binding site and at least one spacer sequence; and(v) a 3’ ITR.

80. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat ( ITR );(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002;(vi) at least one miR183 binding site; and(vii) a 3 ’ ITR.

81. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., al least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical to SEQ ID NO: 1842;(v) the GBA1 -encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001 ;(vi) at least one miR183 binding site: and(vii) a 3 ’ ITR.

82. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GB Al -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence al least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002;(vi) a miR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence at least 90% identical to SEQ ID NO: 1849; and(vii) a 3 ’ ITR.

83. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence SEQ ID NO: 1831 or a nucleotide sequence at least: 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., al least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001 ;(vi) a miR 183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1849; and(vii) a 3 ’ ITR.

84. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO:1829 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g. , at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 93% identical) to SEQ ID NO: 2002;(vi) at least one miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 1847;(vii) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

85. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001;(vi) at least one miRl 83 binding site comprising the nucleotide sequence of SEQ ID NO: 1847;(vii) a polyadenylation (polyA) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) to SEQ ID NO: 1846; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

86. The AAV particle of any one of claims 1-34, wherein the viral genome comprises, in 5’ to 3’ order:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1 -encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2.002 or a nucleotide sequence at least 90% identical (e.g.. at least 93% identical) to SEQ ID NO: 2.002;(vi) a rmR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, al least 92?% at least 93?% at least 94%, at least 95%, at least 96%, al least 97%, at least 98%, or at least 99% identical) io SEQ ID NO: 1849;(vii) a poly adenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) to SEQ ID NO: 1846; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

87. The AAV particle of any one of claims 1 -34, wherein the viral genome comprises, in 5 ’ to 3 ’ order:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1829;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1831;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1834;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) to SEQ ID NO: 1842;(v) the GBA1-encoding sequence, wherein the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or a nucleotide sequence at least 90% identical (e.g., at least 94% identical) to SEQ ID NO: 2001;(vi) a miR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1849;(vii) a polyadenvlation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) to SEQ ID NO: 1846; and(viii) a 3' ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g., at least 95%, al least 96% at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1830.

88. The AAV particle of claim 87, wherein:(i) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829;(ii) the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831;(iii) the CBA promoter comprises the nucleotide sequence of SEQ ID NO: 1834;(iv) the intron comprises the nucleotide sequence of SEQ ID NO: 1842;(v) the GBA1-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001;(vi) the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849;(vii) the poly A region comprises the nucleotide sequence of SEQ ID NO: 1846; and(viii) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

89. The AAV particle of any one of claims 1-34, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., al least 97% identical) to SEQ ID NO: 2007.

90. The AAV particle of claim 89, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007.91 . The AAV particle of claim 90, wherein the viral genome consists of the nucleotide sequence of SEQ ID NO: 2007.

92. An adeno-associated vims (AAV) particle comprising a viral genome comprising the nucleotide sequence of SEQ ID NO: 2.001 and an AAV capsid variant comprising:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:4.

93. An adeno-associated virus (AAV) particle comprising a viral genome comprising the nucleotide sequence of SEQ ID NO: 2001 and an AAV capsid variant comprising:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742. of SEQ ID NO:36; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:36.

94. An adeno-associated virus (AAV) particle comprising a viral genome comprising the nucleotide sequence of SEQ ID NO: 2002 and an AAV capsid variant comprising:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO:4; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 4.

95. An adeno-associated virus (AAV) particle comprising a viral genome comprising the nucleotide sequence of SEQ ID NO: 2002 and an AAV capsid variant comprising:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 36.

96. A ceil comprising the AAV particle of any one of claims 1-95, optionally wherein the cell is a mammalian cell (e.g., an HEK293 ceil), an insect cell (e.g., an Sf9 cell), or a bacterial ceil.

97. A method of making the AAV particle of any one of claims 1-95, the method comprising:(i) providing a cell comprising the viral genome comprising a GBA1 -encoding sequence and a nucleic acid encoding the AAV capsid variant; and(ii) incubating the cell under conditions suitable to encapsulate the viral genome in the AAV capsid variant; thereby making the AAV particle.

98. The method of claim 97, wherein the viral genome comprises(a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; or(b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO: 4;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 138-742 SEQ ID NO: 4; or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identity) to positions 203-742 of SEQ ID NO: 4.

99. The method of claim 97, wherein the viral genome comprises(a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; or(b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 4, the amino acid sequence of positions 138-742 of SEQ ID NO: 4, and / or the amino acid sequence of positions 203-742 of SEQ ID NO: 4.

100. The method of claim 97, wherein the viral genome comprises:(a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; or(b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO: 36;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 138-742 SEQ ID NO: 36; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 203-742 of SEQ ID NO: 36.

101. The method of claim 97, wherein the viral genome comprises:(a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, al least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; or(b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 36, the amino acid sequence of positions 138-742 of SEQ ID NO: 36, and / or the amino acid sequence of positions 203-742 of SEQ ID NO: 36.

102. The method of claim 97, wherein the viral genome comprises:(a) the nucleotide sequence of SEQ ID NO: 2.006 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; or(b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, al least 93%, at least 94%. at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises:(i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO: 982;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742. of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g,, at least 90%, at least 91%, at least 92?% at least 93%, at least 94%, at least 95%, al least 96%, at least 97%, at least 98%, or at least 99% identity ) to positrons 138-742 SEQ ID NO: 982; and / or(iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 203-742 of SEQ ID NO: 982.

103. The method of claim 97, wherein the viral genome comprises:(a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, al least 98%, or at least 99% identical) thereto; or(b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO:

982. the amino acid sequence of positions 138-742 of SEQ ID NO: 982, and / or the amino acid sequence of positions 203-742 of SEQ ID NO: 982.

104. The method of any one of claims 97-103, further comprising, prior to step (i), introducing a first nucleic acid molecule comprising the viral genome into the cell.

105. The method of any one of claims 97-104, wherein the cell comprises a second nucleic acid molecule encoding the AAV capsid variant.

106. The method of claim 105, further comprisi ng, prior to step (i), introducing the second nucleic acid into the cell.

107. The method of any one of claims 97-106. wherein the cell comprises a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an Sf9 cell), or a bacterial cell.

108. A pharmaceutical composition comprising the AAV particle of any one of claims 1-95. and a pharmaceutically acceptable excipient.

109. A pharmaceutical composition comprising the AAV particle of any one of claims 5-22 and a pharmaceutically acceptable excipient.

110. A pliarmaceutical composition comprising the AAV particle of any one of claims 9-15, 92. and 94, and a pharmaceutically acceptable excipient.

111. A pharmacal composition comprising the AAV particle of any one of claims 16-22, 93, and95, and a pharmaceutically acceptable excipient.

112. A method of delivering an AAV particle encoding a GBA1 protein to a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 108-111 or the AAV particle of any one of claims 1-95,113. The method of claim 112, wherein the subject has, lias been diagnosed with having, or is at risk of having a GBA1 -related disorder, optionally wherein the GBA1-related disorder is a GBA1 -related neurodegenerative or neuromuscular disorder, further optionally wherein the GBA1 -related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation, type 1 (NB1A 1), or Hallervorden-Spatz Syndrome.

114. The method of claim 112 or claim 113, wherein the subject has, has been diagnosed with having, or is at risk of having Parkinson’s Disease (PD).

115. A method of treating a subject hatring or diagnosed with having a GBA1-related disorder, comprising administering to the subject an effective amount of the pliarmaceutical composition anyone of claims 108-111 or the AAV particle of any one of claims 1-95.

116. A method of treating a subject having or diagnosed with having a GBA1 -related disorder, comprising administering to the subject an effective amount of the pharmacal composition of any one of claims 109-111 or the AAV particle of any one of claims 5-22.

117. A method of treating a subject having or diagnosed with having a GBA1-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 110 or the AAV particle of any one of claims 9-15, 92, and 94.

118. A method of treating a subject having or diagnosed with having a GBA1-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or the AAV particle of any one of claims 16-22, 93, and 95.

119. The method of any one of claims 115-118, wherein the GBA1 -related disorder is a GBA1-related neurodegenerative or neuromuscular disorder, optionally wherein the GBA1-related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation, type 1 (NBIA 1), or Hallervorden- Spatz Syndrome.

120. The method of claim 119, wherein the GBA1 -related neurodegenerative or neuromuscular disorder is Parkinson’s Disease (PD).

121. The method of claim 119. wherein the GBA1-related neurodegenerative or neuromuscular disorder is Gaucher Disease (GD) (e.g., GD type 1, 2. or 3).

122. The method of claim 121, wherein the GBA1-related neurodegenerative or neuromuscular disorder is GD type 1.

123. The method of claim 121, wherein the GBA1-related neurodegenerative or neuromuscular disorder is GD type 2.

124. The method of claim 121, wherein the GBA1-related neurodegenerative or neuromuscular disorder is GD type 3.

125. The method of claim 119, wherein the GBA1-related neurodegenerative or neuromuscular disorder is Dementia with Lewy Bothes (DLB).

126. The method of claim 119, wherein the GBA1-related neurodegenerative or neuromuscular disorder is Lewy Body Dementia (LBD).

127. A method of treating a subject having Parkinson’s Disease (PD) or diagnosed with having PD, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 108-111 or the AAV particle of any one of claims 1 -95.

128. A method of treating a subject having Parkinson’s Disease (PD) or diagnosed with having PD, comprising administering to the subject an effective amount of the pharmaceutical composition of claim any one of claims 109-111 or the AAV particle of tiny one of claims 5-22.

129. A method of treating a subject having Parkinson’s Disease (PD) or diagnosed with having PD, comprising administering to the subject an effec tive amount of the pharmaceutical composition of claim 110 or the AAV particle of any one of claims 9-15, 92, and 94.

130. A method of treating a subject having Parkinson’s Disease (PD) or diagnosed with having PD, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 1 11 or the AAV particle of any one of claims 16-22, 93, and 95.

131. A method of treating a subject having Gaucher Disease (GD) (e.g., GD type 1 , 2, or 3) or diagnosed with having Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 108-111 or the AAV particle of any one of claims 1-95.

132. A method of treating a subject having Gaucher Disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with having Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 109-111 or the AAV particle of any one of claims 5-22.

133. A method of treating a subject having Gaucher Disease (GD) (e.g., GD type 1, 2, or 3) or diagnosed with having Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 110 or the AAV particle of any one of claims 9-15, 92, and 94.

134. A method of treating a subject having Gaucher Disease (GD) (e.g., GD type 1, 2. or 3) or diagnosed with having Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or the AAV particle of any one of claims 16-22, 93, and 95.

135. The method of any one of claims 131-134, wherein the GD is GD type 1.

136. The method of any one of claims 131-134, wherein the GD is GD type 2.

137. The method of any one of claims 131-134, wherein the GD is GD type 3.

138. A method of treating a subject hatring Lewy Body Dementia (I., ED), comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 108-111 or the AAV particle of any one of claims 1-95.

139. A method of treating a subject having Lewy Body Dementia (LED), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 109-111 or the AAV particle of any one of claims 5-22.

140. A method of treating a subject having Lewy Body Dementia (LED), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or the AAV particle of any one of claims 9-15, 92, and 94.

141. A method of treating a subject having Lewy' Body Dementia (LED), comprising administering to the subject an effective amount of the plrarmacentical composition of claim 111 or the AAV particle of any one of claims 16-22, 93, and 95.

142. A method of treating a subject having Dementia with Lewy Bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 108-111 or the AAV particle of any one of claims 1-95.

143. A method of treating a subject having Dementia with Lewy Bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 109-111 or the AAV particle of any one of claims 5-22.

144. A method of treating a subject having Dementia with Lewy Bodies (DLB), comprising administering to the subject an effective amount of the plrarmacentical composition of claim 111 or the AAV particle of any one of claims 9-15, 92. and 94.

145. A method of treating a subject having Dementia with Lewy Bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 111 or the AAV particle of any one of claims 16-22, 93, and 95.

146. The method of any one of claims 112-145, wherein the subject Iras one or more mutations in the GBA1 gene.

147. The method of any one of claims 112-146, wherein the subject has lower GCase activity as compared to GCase activity in an individual who does not have a GBA1-related disorder, optionally wherein the level of GCase activity is measured by a 4-MUG assay or a SensoLyte Blue Glucocerebrosidase assay.

148. The method of any one of claims 115-147, wherein the treating results in prevention of progression of the disorder in the subject.

149. The method of any one of claims 115-148, wherein the treating results in amelioration of at least one symptom of the disorder and / or a change in one or more biomarkers of the disorder.

150. The method of claim 149. wherein the one or more biomarkers comprises a GCase activity, a level of glucocerebroside and other glycolipids, (e.g.. within immune cells such as macrophages), a level of synuclein aggregates (e.g.. Lewy bodies), a level of neurofilament light chain, or a combination thereof.

151. The method of claim 149. wherein the at least one symptom comprises developmental delay, progressive encephalopathy, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, generalized weakness, trembling of a limb, depression, visual hallucinations, cognitive decline, or a combination thereof.

152. The method of any one of claims 112-151, wherein the subject is a human.

153. The method of any one of claims 112-152, wherein the AAV particle is delivered to a cell, tissue, or region of the CNS, e.g., a region of the brain or spinal cord, e.g., the parenchyma, the cortex, substantia nigra, caudate cerebellum, striatum, corpus callosum, cerebellum, brain stem caudate- putamen. thalamus, superior colliculus, the spinal cord, or a combination thereof, wherein the AAV particle or pharmaceutical composition is delivered via intravenous administration.

154. The method of any one of claims 112-153, further comprising evaluating, e.g., measuring, the level of GBA1 expression, e.g., GBA1 gene expression. GBA1 mRNA expression, and / or GBA1 protein expression, in the subject, e.g., in a cell, tissue, or fluid of the subject.

155. The method of any one of claims 112-154, wherein the level of GBA1 protein expression is measured by an ELISA, a Western blot, or an immunohistochemistry assay.

156. The method of claim 154 or claim 155, wherein evaluating the level of GBA1 expression is performed prior to and subsequent to administration of the AAV, optionally wherein the level of GBA1 expression prior to administration is compared to the level of GBA1 expression subsequent to administration.

157. The method of any one of claims 154-156, comprising evaluating foe level of GBA1 expression in a cell or tissue of the central nervous system (e.g., parenchyma).

158. The method of claim 156 or claim 157, wherein foe subject’s level of GBA1 protein expression subsequent to administration is increased relative to the subject’s level of GBA1 protein expression prior to administration.

159. The method of any one of claims 1 12-158, further comprising evaluating, e.g., measuring, the level of GCase activity in the subject.

160. The method of any one of claims 112-159, wherein the administration results in an increase in:(i) GCase activity in a cell, tissue, (e.g., a cell or tissue of the CNS. e.g., the cortex, striatum, thalamus, cerebellum, and / or brainstem), and / or fluid (e.g.. CSF and / or serum) of the subject, optionally wherein GCase activity in the subject subsequent to the administration is increased by at least 2-fold, at least 3-fold, at least 4-fold, or at least 5-fold relative to GCase activity in the subject prior to the administration;(ii) viral genomes (VG) per ceil level in a CNS tissue (e.g., the cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord) of the subject, optionally wherein the subject’s VG per cell level in the CNS tissue is increased by greater than 50 VG per cell relative to the subject’s VG per cell level in a peripheral tissue; and / or(iii) GBA1 mRNA expression in a cell or tissue (e.g., a cell or tissue of the CNS, e.g., the cortex, thalamus, and / or brainstem) of the subject, optionally wherein GBA1 mRNA expression in the subject subsequent to the administration is increased by at least 100-1300-fold as compared to GBA1 mRNA expression in the subject prior to foe administration.

161. The method of any one of claims 115-160, further comprising administering to the subject an additional agent suitable for treatment or prevention of the GBA1 -related disorder, optionally wherein the additional agent comprises enzyme replacement therapy (ERT) (e.g., imiglucerase, velaglucerase alfa, or tahglucerase alfa); substrate reduction therapy (SRT) (e.g.,elighistat or miglustat), levodopa, carbidopa, Safinamide. a dopamine agonist (e.g., quetiapine, clozapine, pramipexole, rotigotine, or ropinirole), an anticholinergic (e.g., benztropine or trihexyphenidyl), a cholinesterase inhibitor (e.g., rivastigmine, donepezil, or galantamine), an N- methyl-d-aspartate (NMDA receptor antagonist (e.g., memantine), or a combination thereof.

162. The method of any one of claims 112-161, farther comprising administering a blood transfusion to the subject.

163. The method of any one of claims 112-161, further comprising administering an immunosuppressant to the subject.

164. The method of claim 162, wherein the immunosuppressant comprises a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or eculizumab hydroxychloroquine.

165. The pharmaceutical composition of any one of claims 108-111 or the AAV particle of any one of claims 1-95, for use in the treatment of a GBA1-related disorder; optionally wherein the GBA1- related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g.. GD type 1, 2, or 3), Dementia with Lewy' Bodies (LBD), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation, type 1 (NBIA 1), or Hallervorden-Spatz Syndrome.

166. The pharmaceutical composition or the AAV particle of claim 165, wherein the GBA1-related disorder is Gaucher Disease (GD) (e.g., GD type 1, 2, or 3).

167. The pharmaceutical composition or the AAV particle of claim 166, wherein the GD is GD type 1.

168. The pharmaceutical composition or the AAV particle of claim 166, wherein the GD is GD type2.

169. The pharmaceutical composition or the AAV particle of claim 166, wherein the GD is GD type170. The pharmaceutical composition or the AAV particle of claim 165. wherein the GBA1-related disorder is Parkinson’s Disease (PD).

171. The pharmaceutical composition orthe AAV particle of claim 165, wherein the GBA1-related disorder is LBD.

172. The pharmaceutical composition or the AAV particle of claim 165, wherein the GBA1-related disorder is DLB.

173. Use of the pharmaceutical composition of any one of claims 108-111 or the AAV particle of any one of claims 1-95 in the manufacture of a medicament for the treatment of a GBA1-related disorder; optionally wherein the GBA1-related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation, type 1 (NBIA 1), or Hallervorden-Spatz Syndrome.

174. The use of claim 173. wherein the GBA1-related disorder is PD.

175. The use of claim 173, wherein the GBA1 -related disorder is Gaucher Disease (GD) (e.g,, GD type 1, 2, or 3).

176. The use of claim 175, wherein the GD is GD type 1.

177. The use of claim 175, wherein the GD is GD type 2.

178. The use of claim 175, wherein the GD is GD type 3.

179. The use of claim 173, wherein the GBA1-related disorder is LBD.

180. The use of claim 173, wherein the GBA1-related disorder is DLB,