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Updated: Jul 13, 2026
Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro
Published on: February 15, 2022
Proteome analysis of cultivated vascular smooth muscle cells from a CADASIL patient
Saara Ihalainen1, Rabah Soliymani, Erika Iivanainen
1Protein Chemistry Unit, Institute of Biomedicine/Anatomy, University of Helsinki, Helsinki, Finland. saara.ihalainen@helsinki.fi
Insights
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) involves NOTCH3 gene mutations. Proteomic analysis reveals altered proteins in vascular smooth muscle cells, suggesting cellular stress and abnormal contraction contribute to this vascular dementia.
Area of Science:
- Neuroscience
- Genetics
- Cell Biology
Background:
- Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a genetic vascular dementia linked to NOTCH3 gene mutations.
- These mutations often result in altered cysteine residues in the Notch3 receptor's extracellular domain (N3ECD).
- CADASIL pathology involves vascular smooth muscle cell (VSMC) degeneration and N3ECD accumulation in cerebral arteries.
Purpose of the Study:
- To investigate the proteomic profile of genetically confirmed human CADASIL VSMCs.
- To identify differentially expressed proteins and understand their role in CADASIL pathogenesis.
- To explore the cellular mechanisms beyond Notch3 signaling impairment.
Main Methods:
- Proteomic analysis of cultured human CADASIL VSMCs.
- Identification and characterization of differentially expressed proteins.
- Analysis of protein functions related to cellular processes.
Main Results:
- Eleven differentially expressed proteins were identified in CADASIL VSMCs.
- These proteins are implicated in protein degradation, folding, VSMC contraction, and cellular stress.
- Misfolded Notch3 may induce endoplasmic reticulum stress, unfolded protein response, increased reactive oxygen species, and reduced cell proliferation.
- Upregulated contractile proteins suggest altered VSMC contraction signaling.
- Accumulated N3ECD might enhance angiotensin II responsiveness.
Conclusions:
- Protein expression alterations in VSMCs are key in CADASIL, beyond Notch3 signaling defects.
- Cellular stress, protein misfolding, and altered contractility are significant pathological features.
- These findings offer new insights into CADASIL mechanisms and potential therapeutic targets.
Abstract:
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a vascular dementing disease caused by mutations in the NOTCH3 gene, most which are missense mutations leading to an uneven number of cysteine residues in epidermal growth factor-like repeats in the extracellular domain of Notch3 receptor (N3ECD). CADASIL is characterized by degeneration of vascular smooth muscle cells (VSMC) and accumulation of N3ECD on the VSMCs of small and middle-sized arteries. Recent studies have demonstrated that impairment of Notch3 signaling is not the primary cause of the disease. In the present study we used proteomic analysis to characterize the protein expression pattern of a unique material of genetically genuine cultured human CADASIL VSMCs. We identified 11 differentially expressed proteins, which are involved in protein degradation and folding, contraction of VSMCs, and cellular stress. Our findings indicate that misfolding of Notch3 may cause endoplasmic reticulum stress and activation of unfolded protein response, leading to increased reactive oxygen species and inhibition of cell proliferation. In addition, upregulation of contractile proteins suggests an alteration in the signaling system of VSMC contraction. The accumulation of N3ECD on the cell surface possibly upregulates the angiotensin II regulatory feedback loop and thereby enhances the readiness of the cells to respond to angiotensin II stimulation.
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