3D Vessels-on-Chip using isogenic hiPSC-derived VSMCs reveal NOTCH3-driven alterations in brain small vessel disease

Stem Cell Reports, vol. 21, pp. 102863

Abstract

cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a hereditary brain small vessel disease caused by pathogenic variants in the NOTCH3 gene, leading to NOTCH3 protein accumulation and degeneration of vascular smooth muscle cells (VSMCs). Here, we developed a CADASIL 3D Vessel-on-Chip model using either primary brain VSMCs or human induced pluripotent stem cell (hiPSC)-derived VSMCs from CADASIL patients and isogenic controls. In 3D co-culture with hiPSC-derived endothelial cells, both primary and hiPSC-derived CADASIL VSMCs exhibited disease-relevant morphological abnormalities, increased NOTCH3 and contractile protein levels, and altered intracellular Ca 2+ dynamics that were not observed under conventional 2D culture. PDGFRβ, a downstream NOTCH3 target, was upregulated and correlated with NOTCH3 protein levels in both 3D models and CADASIL patient brain tissue. Pharmacological inhibition of NOTCH3 cleavage reduced NOTCH3 protein levels and rescued CADASIL VSMC phenotypic abnormalities. In conclusion, this 3D Vessel-on-Chip model robustly shows CADASIL pathology-relevant readouts and provides a platform for mechanistic studies and therapeutic testing.

Authors 13

1. Marc Vila Cuenca Aachen

   Leiden University Medical Center

   Affiliation as printed

   Department of Clinical Genetics, Leiden University Medical Center, Leiden, the Netherlands; Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, the Netherlands

2. Theano Tsikari Aachen Department of Anatomy and Embryology

   Leiden University Medical Center

   Affiliation as printed

   Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, the Netherlands

3. Minne N. Cerfontaine Aachen Department of Clinical Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Clinical Genetics, Leiden University Medical Center, Leiden, the Netherlands

4. James Gallant Aachen Department of Anatomy and Embryology

   Leiden University Medical Center

   Affiliation as printed

   Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, the Netherlands

5. Francijna E. van den Hil Aachen Department of Anatomy and Embryology

   Leiden University Medical Center

   Affiliation as printed

   Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, the Netherlands

6. Marga J. Bouma Aachen

   Leiden University Medical Center

   Affiliation as printed

   Leiden University Medical Center hiPSC Center, Leiden, the Netherlands

7. Kyra L. Dijkstra Aachen Department of Pathology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Pathology, Leiden University Medical Center, Leiden, the Netherlands

8. Gido Gravesteijn Aachen Department of Clinical Genetics

   Leiden University Medical Center

   Affiliation as printed

   Department of Clinical Genetics, Leiden University Medical Center, Leiden, the Netherlands

9. Antoine A.F. de Vries Aachen Department of Cardiology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands

10. Christine L. Mummery Aachen Department of Anatomy and Embryology

    Leiden University Medical Center

    Affiliation as printed

    Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, the Netherlands

11. Julie W. Rutten corresponding Aachen Department of Clinical Genetics

    Leiden University Medical Center

    Affiliation as printed

    Department of Clinical Genetics, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: j.w.rutten@lumc.nl

12. Saskia A.J. Lesnik Oberstein corresponding Aachen Department of Clinical Genetics

    Leiden University Medical Center

    Affiliation as printed

    Department of Clinical Genetics, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: s.a.m.j.lesnik@lumc.nl

13. Valeria V. Orlova corresponding Aachen Department of Anatomy and Embryology

    Leiden University Medical Center

    Affiliation as printed

    Department of Anatomy & Embryology, Leiden University Medical Center, Leiden, the Netherlands. Electronic address: v.orlova@lumc.nl