A high-throughput screening assay using Krabbe disease patient cells.

[krabbe disease]

Globoid cell leukodystrophy (GLD ) or Krabbe disease is a lysosomal disease caused by β-galactocerebrosidase (GALC ) deficiency resulting in a rapidly progressive neurodegenerative disorder . Unfortunately , the only available treatment is hematopoietic bone marrow transplantation , which prevents its fulminant manifestation but without treating further neurological manifestations . Here , we describe the development of a cellular high -throughput screening (HTS ) assay using GLD patient fibroblasts to screen for small molecules that enhance the residual mutant GALC enzymatic activity . Small molecules have substantial therapeutic potential in GLD because they are more prone to cross the blood -brain barrier , reaching the neuronal affected cells . The transformation of primary skin fibroblasts with SV 40 large T antigen has been shown to maintain the biochemical characteristics of the GLD cells and generates sufficient cells for the HTS . Using a specific fluorescent substrate , residual GALC activity from an SV 40 -transformed GLD patient fibroblast was measurable in high -density microplates . The pilot quantitative HTS against a small compound collection showed robust statistics . The small molecules that showed active concentration-response curves were further studied in primary GLD fibroblasts . This cell-based HTS assay demonstrates the feasibility of employing live GLD patient cells to identify therapeutic agents that can potentially be used for the treatment of this progressive neurodegenerative disease .