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BK Channelopathy in Human Neuronal Models of Angelman Syndrome
Functional studies of 2D human neurons and 3D brain organoids reveal channel dysfunction underlying seizure.
Science article: “Potassium channel dysfunction in human neuronal models of Angelman syndrome” Science. 2019 Dec 20; 366(6472)1486-1492. doi: 10.1126/science.aav5386.
October 28th, 2020
9:30 – 10:30 AM UTC+5.30 Delhi
12:00 – 1:00 PM UTC+8:00 Singapore
9:00 – 10:00 PM UTC-8:00 San Francisco
Alfred Xuyang Sun, PhD
Junior Principal Investigator
National Neuroscience Institute
Duke NUS Medical School
Genome Institute of Singapore
What you will learn:
- Functional human neural cells uncover novel pathophysiological phenotypes
- Confirmation and behavior rescue in animal model
- Potential therapeutic targets suggested
Graphical abstract of Potassium channel dysfunction in human neuronal models of Angelman syndrome scientific paper.
Angelman Syndrome (AS) is a neurodevelopmental disorder characterized by intellectual disability and frequent seizures. Despite clear genetic evidences that most AS cases are caused by loss-of-function mutations or deletions of UBE3A, an E3 ubiquitin ligase, how the network hyperactivity observed in patients arises from UBE3A deficiency in individual neurons is still lacking. Utilizing human neurons and cortical organoids derived from AS patient iPSCs and UBE3A KO hESCs, we observed specific changes in the intrinsic excitability of UBE3A deficient neurons. These changes were caused by augmented fAHP, which is mediated by BK channels. Mechanistically, we demonstrated UBE3A-mediated ubiquitination and proteasomal degradation of BK. Overall, our integrative analysis of data obtained from both 2D neurons and 3D organoids using a combination of genetic, biochemical, electrophysiological, and imaging assays including those imaged under Azure c600 Imaging System, uncovers a new pathophysiological mechanism underlying the hyperexcitability of AS, posits AS as a channelopathy syndrome, and identifies a novel substrate of UBE3A that is amendable for pharmacological interventions.
Representative data of Azure c600 Imaging system
Fig. 1 (A). Schematic illustrating the CRISPR-Cas9-mediated gene editing approach used to knock out UBE3A in hESCs (top) and immunoblot showing the absence of UBE3A protein in UBE3A KO hESCs (bottom).
Data contribution in this scientific finding:
Azure c600 Imaging system visualized the detected chemiluminescence and NIR fluorescence signals on immunoblots and co-immunoprecipitation assay. The data validated and quantified various protein levels including neuronal markers in BK- and UEB3A-overexpressed HEK293 cells, WT and UEB3A-KO human neurons and brain organoids (Fig. 1 (A), Fig. S3 (E), Fig. S6, Fig. S10 (D), Fig. S13).
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