A Strategy for Treatment of Gaucher Disease via Enzyme Replacement Therapy

Dutta, Tahiti (2023) A Strategy for Treatment of Gaucher Disease via Enzyme Replacement Therapy. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Tahiti Dutta (18MS165)) Thesis_18MS165.pdf - Submitted Version Restricted to Repository staff only Download (6MB)

Abstract

Gaucher Disease (GD) is a type of Lysosomal Storage Disorder (LSD) caused due to reduced activity of β-glucocerebrosidase (GCase) and accumulation glucosylceramide in lysosomes, finally resulting in necrotic failures, multi-organ damage and neurodegenerative disorders. Here, we discuss an Enzyme Replacement Therapy (ERT) approach to effectively treat GD by covalently labelling B8CYA8, an enzyme of the family of GCase extracted from a thermophilic bacteria, Halothermothrix orenii, with a mannose-6-phosphate functionalized polylysine glycopolypeptide and specifically deliver it to lysosome by receptor mediated endocytosis employing the Cation Independent Mannose-6-Phosphate Receptor (CIM6PR). The G306 residue of B8CYA8 was mutated to introduce a cysteine residue on the surface, which was used for thiol-maleimide coupling to attach a pH responsive linker and an azide group. The propargyl functionalized glucopolypeptide will be “clicked” with the azide using Copper- Assisted Azide-Alkyne Cycloaddition (CuAAC). In this thesis, the optimization of the bioconjugation reactions are discussed. The activity of the conjugated protein was compared with the wild type and sufficient retention in activity was found.

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