Cellular Functions and Gene and Protein Expression Profiles in Endothelial Cells Derived from Moyamoya Disease-Specific iPS Cells.
OBJECTIVE
Moyamoya disease (MMD) is a slow, progressive steno-occlusive disease, arising in the terminal portions of the cerebral internal carotid artery. However, the functions and characteristics of the endothelial cells (ECs) in MMD are unknown. We analyzed these features using induced pluripotent stem cell (iPSC)-derived ECs.


METHODS
iPSC lines were established from the peripheral blood of three patients with MMD carrying the variant RNF213 R4810K, and three healthy persons used as controls. After the endothelial differentiation of iPSCs, CD31+CD144+ cells were purified as ECs using a cell sorter. We analyzed their proliferation, angiogenesis, and responses to some angiogenic factors, namely VEGF, bFGF, TGF-β, and BMP4. The ECs were also analyzed using DNA microarray and proteomics to perform comprehensive gene and protein expression analysis.


RESULTS
Angiogenesis was significantly impaired in MMD regardless of the presence of any angiogenic factor. On the contrary, endothelial proliferation was not significant between control- and MMD-derived cells. Regarding DNA microarray, pathway analysis illustrated that extracellular matrix (ECM) receptor-related genes, including integrin β3, were significantly downregulated in MMD. Proteomic analysis revealed that cytoskeleton-related proteins were downregulated and splicing regulation-related proteins were upregulated in MMD.


CONCLUSIONS
Downregulation of ECM receptor-related genes may be associated with impaired angiogenic activity in ECs derived from iPSCs from patients with MMD. Upregulation of splicing regulation-related proteins implied differences in splicing patterns between control and MMD ECs.
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Shuji
S
Hamauchi
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Hideo
H
Shichinohe
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Haruto
H
Uchino
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Shigeru
S
Yamaguchi
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Naoki
N
Nakayama
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Ken
K
Kazumata
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Toshiya
T
Osanai
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Takeo
T
Abumiya
Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Kiyohiro
K
Houkin
Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
Takumi
T
Era
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