Bugallo-Casal Ana, Muiño Elena, Bravo Susana B, Hervella Pablo, Arias-Rivas Susana, Rodríguez-Yáñez Manuel, Vara-León Enrique, Quintas-Rey Rita, Pérez-Gayol Lara, Maisterra-Santos Olga, Pizarro-Gonzálvez Jesús, Martorell-Riera María Rosa, Vives-Bauzá Cristòfol, Fernández-Cadenas Israel, Castillo José, Campos Francisco
Translational Stroke Laboratory Group (TREAT), Clinical Neurosciences Research Laboratory (LINC), Health Research Institute of Santiago de Compostela (IDIS), 15706, Santiago de Compostela, Spain.
University of Santiago de Compostela (USC), 15705, Santiago de Compostela, Spain.
Neuromolecular Med. 2025 Feb 27;27(1):18. doi: 10.1007/s12017-025-08840-6.
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common genetic form of stroke. It is caused by a cysteine-altering variant in one of the 34 epidermal growth factor-like repeat (EGFr) domains of Notch3. NOTCH3 pathogenic variants in EGFr 1-6 are associated with high disease severity, whereas those in EGFr 7-34 are associated with late stroke onset and increased survival. However, whether and how the position of the NOTCH3 variant directly affects the disease severity remains unclear. In this study, we aimed to generate human-induced pluripotent stem cells (hiPSCs) from patients with CADASIL with EGFr 1-6 and 7-34 pathogenic variants to evaluate whether the NOTCH3 position affects the cell phenotype and protein profile of the generated hiPSCs lines. Six hiPSCs lines were generated: two from patients with CADASIL with EGFr 1-6 pathogenic variants, two from patients with EGFr 7-34 variants, and two from controls. Notch3 aggregation and protein profiles were tested in the established six hiPSCs lines. Cell analysis revealed that the NOTCH3 variants did not limit the cell reprogramming efficiency. However, EGFr 1-6 variant position was associated with increased accumulation of Notch3 protein in pluripotent stem cells and proteomic changes related with cytoplasmic reorganization mechanisms. In conclusion, our analysis of hiPSCs derived from patients with CADASIL support the clinical association between the NOTCH3 variant position and severity of CADASIL.
伴有皮质下梗死和白质脑病的常染色体显性遗传性脑动脉病（CADASIL）是最常见的遗传性卒中形式。它由Notch3的34个表皮生长因子样重复序列（EGFr）结构域之一中的半胱氨酸改变变体引起。EGFr 1 - 6中的NOTCH3致病变体与高疾病严重程度相关，而EGFr 7 - 34中的变体与卒中发病较晚和生存率增加相关。然而，NOTCH3变体的位置是否以及如何直接影响疾病严重程度仍不清楚。在本研究中，我们旨在从具有EGFr 1 - 6和7 - 34致病变体的CADASIL患者中生成人诱导多能干细胞（hiPSC），以评估NOTCH3位置是否影响所生成的hiPSC系的细胞表型和蛋白质谱。生成了6个hiPSC系：两个来自具有EGFr 1 - 6致病变体的CADASIL患者，两个来自具有EGFr 7 - 34变体的患者，以及两个来自对照。在已建立的6个hiPSC系中测试了Notch3聚集和蛋白质谱。细胞分析显示，NOTCH3变体并不限制细胞重编程效率。然而，EGFr 1 - 6变体位置与多能干细胞中Notch3蛋白的积累增加以及与细胞质重组机制相关的蛋白质组学变化有关。总之，我们对来自CADASIL患者的hiPSC的分析支持了NOTCH3变体位置与CADASIL严重程度之间的临床关联。
