Modeling CADASIL vascular pathologies with patient-derived induced pluripotent stem cells

Chen Ling,Lina Fu,Jing Qu,Moshi Song,Si Wang,Shuai Ma,Qun Chu,Zhaoxia Wang,Zunpeng Liu,Juan Carlos Izpisua Belmonte,Guang-Hui Liu,Weiqi Zhang,Yun Yuan,Xiaoqian Liu,Shuhui Sun

doi:10.1007/s13238-019-0608-1

Abstract

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a rare hereditary cerebrovascular disease caused by a NOTCH3 mutation. However, the underlying cellular and molecular mechanisms remain unidentified. Here, we generated non-integrative induced pluripotent stem cells (iPSCs) from fibroblasts of a CADASIL patient harboring a heterozygous NOTCH3 mutation (c.3226C>T, p.R1076C). Vascular smooth muscle cells (VSMCs) differentiated from CADASIL-specific iPSCs showed gene expression changes associated with disease phenotypes, including activation of the NOTCH and NF-κB signaling pathway, cytoskeleton disorganization, and excessive cell proliferation. In comparison, these abnormalities were not observed in vascular endothelial cells (VECs) derived from the patient’s iPSCs. Importantly, the abnormal upregulation of NF-κB target genes in CADASIL VSMCs was diminished by a NOTCH pathway inhibitor, providing a potential therapeutic strategy for CADASIL. Overall, using this iPSC-based disease model, our study identified clues for studying the pathogenic mechanisms of CADASIL and developing treatment strategies for this disease.

Highlights

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a hereditary cerebrovascular disease caused by a NOTCH3 gene mutation (Joutel et al, 1996; Goate and Morris, 1997; Rutten et al, 2014), has the clinical manifestations of recurrent ischemic stroke, progressive cognitive decline and mental disorders (Wang et al, 2011; Di Donato et al, 2017; Fang et al, 2017)
Heterozygous mutations of the NOTCH3 gene (c.3226C>T, p.R1076C) in CADASIL fibroblasts and induced pluripotent stem cells (iPSCs) were verified via genomic PCR and sequencing (Fig. 1B)
No significant difference in reprogramming efficiency was observed between WT and CADASIL fibroblasts, and no integrated foreign genes were detected in any of the three iPSC lines (Fig. S1A and S1B)

Summary

Introduction

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a hereditary cerebrovascular disease caused by a NOTCH3 gene mutation (Joutel et al, 1996; Goate and Morris, 1997; Rutten et al, 2014), has the clinical manifestations of recurrent ischemic stroke, progressive cognitive decline and mental disorders (Wang et al, 2011; Di Donato et al, 2017; Fang et al, 2017). Abnormalities in proliferation ability, mitochondrial function and cytoskeleton structure have been identified in VSMCs from CADASIL patients and mice (Domenga et al, 2004; Tikka et al, 2012; Viitanen et al, 2013; Panahi et al, 2018). Despite these prior studies, detailed phenotypic profiles of VSMCs and other types of cells in CADASIL patients, such as VECs, and the underlying mechanism of CADASIL remain elusive

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Results

Conclusion