Yuki Maeda1,2, Keiichi Motoyama1, Taishi Higashi1, Yuka Horikoshi3, Toru Takeo3, Naomi Nakagata3, Yuki Kurauchi1,2, Hiroshi Katsuki1, Yoichi Ishitsuka1, Yuki Kondo1, Tetsumi Irie1,2, Hirokazu Furuya4, Takumi Era5, Hidetoshi Arima1,2. 1. Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan. 2. Program for Leading Graduate Schools 'HIGO (Health Life Science: Interdisciplinary and Glocal Oriented) Program', Kumamoto University, Kumamoto, Japan. 3. Division of Reproductive Engineering, Center for Animal Resources and Development, Kumamoto University, Kumamoto, Japan. 4. Department of Neurology, Kochi Medical School, Kochi University, Kochi, Japan. 5. Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
Abstract
OBJECTIVES: GM1-gangliosidosis is an inherited disorder characterized by the accumulation of GM1-gangliosides in many tissues and organs, particularly in the brain. Currently, there is no treatment available for patients with ganglioside storage diseases. Therefore, we investigated the effects of cyclodextrins (CyDs) on the GM1-ganglioside level in EA1 cells, fibroblasts from patients with GM1-gangliosidosis. METHODS: The concentrations of cholesterol and phospholipids in supernatants were determined by Cholesterol E-test Wako and Phospholipid C-test Wako, respectively. The effects of CyDs on GM1-ganglioside levels in EA1 cells using fluorescence-labelled cholera toxin B-subunit, which can bind to GM1-gangliosides specifically, were investigated by flow cytometry and confocal laser scanning microscopy. KEY FINDINGS: The treatment with methylated CyDs, hydroxypropylated CyDs and branched CyDs decreased GM1-ganglioside levels in EA1 cells at 1 mm for 24 h. Unexpectedly, there was no significant change in the efflux of cholesterol or phospholipids from the cells after treatment with CyDs under the same experimental conditions, indicating that the efflux of membrane components is not associated with down-regulation of GM1-ganglioside levels in EA1 cells upon CyDs treatment. CONCLUSIONS: CyDs may have the potential as drugs for GM1-gangliosidosis, although the mechanism should be thereafter clarified.
OBJECTIVES:GM1-gangliosidosis is an inherited disorder characterized by the accumulation of GM1-gangliosides in many tissues and organs, particularly in the brain. Currently, there is no treatment available for patients with ganglioside storage diseases. Therefore, we investigated the effects of cyclodextrins (CyDs) on the GM1-ganglioside level in EA1 cells, fibroblasts from patients with GM1-gangliosidosis. METHODS: The concentrations of cholesterol and phospholipids in supernatants were determined by Cholesterol E-test Wako and Phospholipid C-test Wako, respectively. The effects of CyDs on GM1-ganglioside levels in EA1 cells using fluorescence-labelled cholera toxin B-subunit, which can bind to GM1-gangliosides specifically, were investigated by flow cytometry and confocal laser scanning microscopy. KEY FINDINGS: The treatment with methylated CyDs, hydroxypropylated CyDs and branched CyDs decreased GM1-ganglioside levels in EA1 cells at 1 mm for 24 h. Unexpectedly, there was no significant change in the efflux of cholesterol or phospholipids from the cells after treatment with CyDs under the same experimental conditions, indicating that the efflux of membrane components is not associated with down-regulation of GM1-ganglioside levels in EA1 cells upon CyDs treatment. CONCLUSIONS:CyDs may have the potential as drugs for GM1-gangliosidosis, although the mechanism should be thereafter clarified.
Authors: Serena Monaco; Samuel Walpole; Hassan Doukani; Ridvan Nepravishta; Macarena Martínez-Bailén; Ana T Carmona; Javier Ramos-Soriano; Maria Bergström; Inmaculada Robina; Jesus Angulo Journal: Chemistry Date: 2020-07-20 Impact factor: 5.236