Lysosomal protein targeting sequence and therapeutic applications of same
The present disclosure relates to methods of treating diseases states such as lysosomal storage diseases and/or neurodegenerative diseases. Also disclosed are one or more compositions that may be useful for one or more of the disclosed methods, including compositions that may comprise acid β-glucosidase (GCase) protein comprising one or more mutations, peptides of acid β-glucosidase, and DNA vectors and cell lines related to acid β-glucosidase peptides or proteins.
1. A glucocerebrosidase (GCase) comprising at least 85% sequence identity to human GCase amino acid sequence as set forth in SEQ ID NO: 2 comprising one or more mutations at a position selected from 397, 399, 400, 401, 402, 403, 407, 408, 409, and combinations thereof, wherein said mutation decreases LIMP-2 binding, wherein the residues at positions 399-403 with reference to SEQ ID NO: 2 comprise a sequence selected from ASPII (SEQ ID NO: 7), DSAII (SEQ ID NO: 8), DSPAI (SEQ ID NO: 9), DSPIA (SEQ ID NO: 10), ASPIA (SEQ ID NO: 11), ASPAA (SEQ ID NO: 12), ESAII (SEQ ID NO: 21), ESPAI (SEQ ID NO: 22), ESPIA (SEQ ID NO: 23), and wherein said GCase protein at position 407-409 of SEQ ID NO: 2 comprises a sequence selected from AAA, AKA, TAD, and TAE.
2. The GCase protein of claim 1 , wherein said mutation substantially ablates LIMP-2 binding.
3. The GCase protein of claim 1 , wherein said mutation changes the charge at said position.
4. The GCase protein of claim 1 , wherein said mutation changes the charge at said position to a positive charge.
5. The GCase protein of claim 1 , wherein said mutation changes the amino acid at said position to an amino acid selected from alanine and glycine.
6. The GCase protein of claim 1 , wherein said position is selected from 399, 402, 403, 407, 408, 409, and a combination thereof.
7. The GCase protein of claim 1 , wherein said position is selected from 399, 402, 403, and a combination thereof.
8. The GCase protein of claim 1 , wherein said position is selected from 407, 408, 409, and a combination thereof.
9. The GCase protein of claim 1 , wherein said mutation decreases localization of said recombinant protein to the lysosomes of a cell while it is synthesized.
10. The GCase protein of claim 1 , wherein said GCase protein has biological activity that is substantially equivalent to the wild type GCase biological activity.
11. The GCase protein of claim 1 , wherein said GCase protein has enzymatic activity that is substantially equivalent to the wild type GCase biological activity.
12. The GCase protein of claim 1 , wherein said GCase protein is capable of being taken up into cells via non-LIMP-2 mechanisms in substantially the same manner as wild type GCase.
13. The GCase protein of claim 1 , wherein said GCase protein further comprises additional N-terminal and/or C-terminal amino acids.
14. The GCase protein of claim 1 , wherein said mutation increases secretion of said GCase protein from cells in which it is synthesized.
15. The GCase protein of claim 1 , comprising a mutation selected from D399E, D405E, D409E, and combinations thereof.