Cell-based high-throughput screening identifies galactocerebrosidase enhancers as potential small-molecule therapies for Krabbe's disease

Dae Song Jang, Wenjuan Ye, Tian Guimei, Melani Solomon, Noel Southall, Xin Hu, Juan Marugan, Marc Ferrer, Gustavo H B Maegawa

Research output: Contribution to journalArticle

Abstract

Krabbe's disease, also known as globoid cell leukodystrophy (GLD), is a lysosomal storage disease caused by the deficiency of the lysosomal enzyme β-galactocerebrosidase (GALC), resulting in severe neurological manifestations related to demyelination secondary to elevated galactosylsphingosine (psychosine) with its subsequent cytotoxicity. The only available treatment is hematopoietic stem cell transplantation, which delays disease onset but does not prevent long-term neurological manifestations. This article describes the identification of small molecules that enhance mutant GALC activity, identified by quantitative cell-based high-throughput screening (qHTS). Using a specific neurologically relevant murine cell line (145M-Twi) modified to express common human hGALC-G270D mutant, we were able to detect GALC activity in a 1,536-well microplate format. The qHTS of approximately 46,000 compounds identified three small molecules that showed significant enhancements of residual mutant GALC activity in primary cell lines from GLD patients. These compounds were shown to increase the levels of GALC-G270D mutant in the lysosomal compartment. In kinetic assessments, these small molecules failed to disturb the GALC kinetic profile under acidic conditions, which is highly desirable for folding-assisting molecules operating in the endoplasmic reticulum and not affecting GALC catalytic properties in the lysosomal compartment. In addition, these small molecules rescued the decreased GALC activity at neutral pH and partially stabilized GALC under heat-denaturating conditions. These drug-like compounds can be used as the starting point to develop novel small-molecule agents to treat the progressive neurodegenerative course of GLD.

Original languageEnglish (US)
Pages (from-to)1231-1245
Number of pages15
JournalJournal of Neuroscience Research
Volume94
Issue number11
DOIs
StatePublished - Nov 1 2016
Externally publishedYes

Fingerprint

Galactosylceramidase
Globoid Cell Leukodystrophy
Psychosine
Therapeutics
Neurologic Manifestations
Lysosomal Storage Diseases
Cell Line
Hematopoietic Stem Cell Transplantation
Demyelinating Diseases
Endoplasmic Reticulum
Hot Temperature

Keywords

  • Krabbe's disease
  • quantitative high-throughput screening
  • small molecules
  • β-galactocerebrosidase

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this

Cell-based high-throughput screening identifies galactocerebrosidase enhancers as potential small-molecule therapies for Krabbe's disease. / Jang, Dae Song; Ye, Wenjuan; Guimei, Tian; Solomon, Melani; Southall, Noel; Hu, Xin; Marugan, Juan; Ferrer, Marc; Maegawa, Gustavo H B.

In: Journal of Neuroscience Research, Vol. 94, No. 11, 01.11.2016, p. 1231-1245.

Research output: Contribution to journalArticle

Jang, DS, Ye, W, Guimei, T, Solomon, M, Southall, N, Hu, X, Marugan, J, Ferrer, M & Maegawa, GHB 2016, 'Cell-based high-throughput screening identifies galactocerebrosidase enhancers as potential small-molecule therapies for Krabbe's disease', Journal of Neuroscience Research, vol. 94, no. 11, pp. 1231-1245. https://doi.org/10.1002/jnr.23875
Jang, Dae Song ; Ye, Wenjuan ; Guimei, Tian ; Solomon, Melani ; Southall, Noel ; Hu, Xin ; Marugan, Juan ; Ferrer, Marc ; Maegawa, Gustavo H B. / Cell-based high-throughput screening identifies galactocerebrosidase enhancers as potential small-molecule therapies for Krabbe's disease. In: Journal of Neuroscience Research. 2016 ; Vol. 94, No. 11. pp. 1231-1245.
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AU - Hu, Xin

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AU - Ferrer, Marc

AU - Maegawa, Gustavo H B

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