An iPSC-based neural model of sialidosis uncovers glycolytic impairment-causing presynaptic dysfunction and deregulation of Ca2+ dynamics

Haruki Odaka; Tadahiro Numakawa; Minami Soga; Jun Kido; Shiro Matsumoto; Ryutaro Kajihara; Toshika Okumiya; Naoki Tani; Yuki Tanoue; Takaichi Fukuda; Hirokazu Furuya; Takafumi Inoue; Takumi Era

doi:10.1016/j.nbd.2021.105279

An iPSC-based neural model of sialidosis uncovers glycolytic impairment-causing presynaptic dysfunction and deregulation of Ca²⁺ dynamics

Neurobiol Dis. 2021 May:152:105279. doi: 10.1016/j.nbd.2021.105279. Epub 2021 Jan 29.

Authors

Haruki Odaka¹, Tadahiro Numakawa², Minami Soga², Jun Kido³, Shiro Matsumoto³, Ryutaro Kajihara⁴, Toshika Okumiya⁴, Naoki Tani⁵, Yuki Tanoue⁶, Takaichi Fukuda⁷, Hirokazu Furuya⁸, Takafumi Inoue⁹, Takumi Era¹⁰

Affiliations

¹ Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan; Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, Ibaraki, Japan.
² Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
³ Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan; Department of Pediatrics, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan.
⁴ Department of Biomedical Laboratory Sciences, Faculty of Life Science, Kumamoto University, Kumamoto, Japan.
⁵ Liaison Laboratory Research Promotion Center, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
⁶ International Research Center for Medical Sciences, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
⁷ Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan.
⁸ Department of Neurology, Kochi Medical School, Kochi University, Kochi 783-8505, Japan.
⁹ Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, Tokyo, Japan.
¹⁰ Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan. Electronic address: tera@kumamoto-u.ac.jp.

PMID: 33516873
DOI: 10.1016/j.nbd.2021.105279

Abstract

Sialidosis is a neuropathic lysosomal storage disease caused by a deficiency in the NEU1 gene-encoding lysosomal neuraminidase and characterized by abnormal accumulation of undigested sialyl-oligoconjugates in systemic organs including brain. Although patients exhibit neurological symptoms, the underlying neuropathological mechanism remains unclear. Here, we generated induced pluripotent stem cells (iPSCs) from skin fibroblasts with sialidosis and induced the differentiation into neural progenitor cells (NPCs) and neurons. Sialidosis NPCs and neurons mimicked the disease-like phenotypes including reduced neuraminidase activity, accumulation of sialyl-oligoconjugates and lysosomal expansions. Functional analysis also revealed that sialidosis neurons displayed two distinct abnormalities, defective exocytotic glutamate release and augmented α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate receptor (AMPAR)-mediated Ca²⁺ influx. These abnormalities were restored by overexpression of the wild-type NEU1 gene, demonstrating causative role of neuraminidase deficiency in functional impairments of disease neurons. Comprehensive proteomics analysis revealed the significant reduction of SNARE proteins and glycolytic enzymes in synaptosomal fraction, with downregulation of ATP production. Bypassing the glycolysis by treatment of pyruvate, which is final metabolite of glycolysis pathway, improved both the synaptsomal ATP production and the exocytotic function. We also found that upregulation of AMPAR and L-type voltage dependent Ca²⁺ channel (VDCC) subunits in disease neurons, with the restoration of AMPAR-mediated Ca²⁺ over-load by treatment of antagonists for the AMPAR and L-type VDCC. Our present study provides new insights into both the neuronal pathophysiology and potential therapeutic strategy for sialidosis.

Keywords: AMPA receptor; Exocytosis; Glycolysis; Lysosomal storage disease; Neuraminidase; Neurotransmission; SNARE protein; Sialidosis; Voltage dependent Ca(2+) channel; iPSC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calcium Signaling / physiology*
Exocytosis / physiology
Glycolysis / physiology
Humans
Induced Pluripotent Stem Cells
Mucolipidoses / physiopathology*
Neurons / pathology*
Neurons / physiology*
Synapses / pathology
Synapses / physiology

Supplementary concepts

Neuraminidase 1 deficiency