학술논문
Development of a Patient-Derived Induced Pluripotent Stem Cell Model for the Investigation of SCN5A-D1275N-Related Cardiac Sodium Channelopathy
Document Type
Journal Article
Author
Fumika Yokoi; Futoshi Toyoda; Hirohiko Kohjitani; Hiroshi Watanabe; Jiarong Chen; Kazuhisa Chonabayashi; Kenichi Sasaki; Koh Ono; Mamoru Hayano; Minoru Horie; Osamu Baba; Sayako Hirose; Seiko Ohno; Shunsuke Funakoshi; Suguru Nishiuchi; Takahiro Horie; Takeru Makiyama; Takeshi Harita; Takeshi Kimura; Tsukasa Kamakura; Yimin Wuriyanghai; Yoshinori Yoshida; Yuta Yamamoto
Source
Circulation Journal. 2017, 81(12):1783
Subject
Language
English
ISSN
1346-9843
1347-4820
1347-4820
Abstract
Methods and Results:First, using a human embryonic kidney (HEK) 293 cell-based heterologous expression system, theSCN5A-D1275N channels showed similar maximum sodium conductance but a significantly depolarizing shift of activation gate (+10 mV) compared to wild type. Second, we generated human-induced pluripotent stem cells (hiPSCs) from a 24-year-old female who carried heterozygousSCN5A-D1275N and analyzed the differentiated cardiomyocytes (CMs). AlthoughSCN5Atranscript levels were equivalent between D1275N and control hiPSC-CMs, both the total amount of NaV1.5 and the membrane fractions were reduced approximately half in the D1275N cells, which were rescued by the proteasome inhibitor MG132 treatment. Electrophysiological assays revealed that maximum sodium conductance was reduced to approximately half of that in control hiPSC-CMs in the D1275N cells, and maximum upstroke velocity of action potential was lower in D1275N, which was consistent with the reduced protein level of NaV1.5.