The moyamoya disease susceptibility variant RNF213 R4810K (rs112735431) induces genomic instability by mitotic abnormality

作者：Hitomi Toshiaki; Habu Toshiyuki; Kobayashi Hatasu; Okuda Hiroko; Harada Kouji H; Osafune Kenji; Taura Daisuke; Sone Masakatsu; Asaka Isao; Ameku Tomonaga; Watanabe Akira; Kasahara Tomoko; Sudo Tomomi; Shiota Fumihiko; Hashikata Hirokuni; Takagi Yasushi; Morito Daisuke; Miyamoto Susumu; Nakao Kazuwa; Koizumi Akio^*

来源：Biochemical and Biophysical Research Communications, 2013, 439(4): 419-426.

DOI：10.1016/j.bbrc.2013.08.067

• Moyamoya disease
• iPS cells
• Mitotic phase
• Genomic instability
• Rs112735431
• MAD2

Moyamoya disease (MMD) is a cerebrovascular disease characterized by occlusive lesions in the Circle of Willis. The RNF213 R4810K polymorphism increases susceptibility to MMD. In the present study, we characterized phenotypes caused by overexpression of RNF213 wild type and R4810K variant in the cell cycle to investigate the mechanism of proliferation inhibition. Overexpression of RNF213 R48101K in HeLa cells inhibited cell proliferation and extended the time of mitosis 4-fold. Ablation of spindle checkpoint by depletion of mitotic arrest deficiency 2 (MAD2) did not shorten the time of mitosis. Mitotic morphology in HeLa cells revealed that MAD2 colocalized with RNF213 R4810K. Immunoprecipitation revealed an RNF213/MAD2 complex: R48101K formed a complex with MAD2 more readily than RNF213 wild-type. Desynchronized localization of MAD2 was observed more frequently during mitosis in fibroblasts from patients (n = 3, 61.0 +/- 8.2%) compared with wild-type subjects (n = 6, 13.1 +/- 7.7%; p < 0.01). Aneuploidy was observed more frequently in fibroblasts (p < 0.01) and induced pluripotent stem cells (iPSCs) (p < 0.03) from patients than from wild-type subjects. Vascular endothelial cells differentiated from iPSCs (iPSECs) of patients and an unaffected carrier had a longer time from prometaphase to metaphase than those from controls (p < 0.05). iPSECs from the patients and unaffected carrier had significantly increased mitotic failure rates compared with controls (p < 0.05). Thus, RNF213 R4810K induced mitotic abnormalities and increased risk of genomic instability.

• 出版日期2013-10-4