Plant cell culture expressing human lysosomal proteins and uses thereof
A device, system and method for producing glycosylated proteins in plant culture, particularly proteins having a high mannose glycosylation, while targeting such proteins with an ER signal and/or by-passing the Golgi. The invention further relates to vectors and methods for expression and production of enzymatically active high mannose lysosomal enzymes using transgenic plant root, particularly carrot cells. More particularly, the invention relates to host cells, particularly transgenic suspended carrot cells, vectors and methods for high yield expression and production of biologically active high mannose Glucocerebrosidase (GCD). The invention further provides for compositions and methods for the treatment of lysosomal storage diseases.
1. A method of increasing hydrolysis of glycosphingolipid glucocerebroside in macrophages of a subject in need thereof, the method comprising administering to said subject a therapeutically effective amount of lyophilized plant cells expressing a recombinant human glucocerebrosidase protein comprising the amino acid sequence of SEQ ID NO: 15, thereby increasing hydrolysis of glycosphingolipid glucocerebroside in macrophages of the subject.
2. The method of claim 1 , wherein said human glucocerebrosidase protein is glycosylated and comprises at least one exposed mannose, at least one fucose having an alpha (1-3) glycosidic bond and at least one xylose.
3. The method of claim 1 , wherein said human glucocerebrosidase protein is linked at its N terminus to an endoplasmic reticulum signal peptide.
4. The method of claim 1 , wherein said glucocerebrosidase protein has glucocerebrosidase catalytic activity.
5. The method of claim 1 , wherein said plant cells are carrot cells from suspension culture.
6. The method of claim 1 wherein said plant cells are formulated in a pharmaceutical composition for treating Gaucher's disease comprising the plant cells and a pharmaceutically acceptable carrier.
7. The method of claim 6 , wherein said pharmaceutical composition is formulated for oral administration.