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Published on: February 15, 2022
Vascular Smooth Muscle Cell Derived from IPS Cell of Moyamoya Disease - Comparative Characterization with Endothelial
Kikutaro Tokairin1, Shuji Hamauchi1, Masaki Ito2
1Department of Neurosurgery, Graduate School of Medicine, Hokkaido University, Sapporo, Japan, 060-8638.
Background:
Moyamoya disease (MMD) is an occlusive cerebrovascular disease, causing stroke in children and young adults with unknown etiology. The fundamental pathology is fibrocellular intimal thickening of cerebral arteries, in which vascular smooth muscle cells (VSMCs) are observed as one of the major cell types. Although the characteristics of circulating smooth muscle progenitor cells have been previously reported, the VSMCs are poorly characterized in MMD. We aimed to characterize VSMCs in MMD using induced pluripotent stem cell (iPSC)-technology.
Methods:
We differentiated VSMCs from neural crest stem cells (NCSCs) using peripheral blood mononuclear cell-derived iPSCs and compared biological and transcriptome features under naïve culture conditions between three independent healthy control (HC) subjects and three MMD patients. VSMC transcriptome profiles were also compared to those of endothelial cells (ECs) differentiated from the same iPSCs.
Results:
Homogeneous spindle-shaped cells differentiated from iPSCs exhibited smooth muscle cell marker expressions, including α-smooth muscle actin (αSMA, 82.3 ± 6.7% and 81.0 ± 6.7%); calponin (91.3 ± 2.1% and 90.9 ± 1.3%); myosin heavy chain-11 (MYH11, 96.9 ± 0.7% and 97.1 ± 0.3%) without significance of differences between the two groups. Real-time PCR showed few PECAM1 and CD34 gene expressions in both groups, indicating features of differentiated VSMCs. There were no significant differences in cellular proliferation (p = 0.45), migration (p = 0.60), and contractile abilities (p = 0.96) between the two groups. Transcriptome analysis demonstrated similar gene expression profiles of VSMCs in HC subjects and MMD patients with six differentially expressed genes (DEGs); while ECs showed a distinct transcriptome profile in MMD patients with 120 DEGs. The Wnt-signaling pathway was a significant pathway in VSMCs.
Conclusions:
This is the first study that established VSMCs from NCSCs using MMD patient-derived iPSCs and demonstrated similar biological function and transcriptome profile of iPSC-derived VMSCs in MMD patients and HC subjects under naïve single culture condition. Comparative transcriptome features between iPSC-derived VSMCs and ECs, displaying distinct transcriptome in the ECs, suggested that pathological traits can be driven by naïve ECs predominantly and VSMCs may require specific environmental factors in MMD, which provides novel insight into the pathophysiology of MMD. Our iPSC derived VSMC model can contribute to further investigations of diagnostic and therapeutic target of MMD in addition to the current iPSC derived EC model.
Insights
Vascular smooth muscle cells (VSMCs) from Moyamoya disease (MMD) patients and healthy controls showed similar functions and gene expression. Endothelial cells (ECs) in MMD patients exhibited distinct transcriptome profiles, suggesting ECs may drive MMD pathology.
Area of Science:
- Stem cell biology
- Cerebrovascular diseases
- Genomics
Background:
- Moyamoya disease (MMD) is an occlusive cerebrovascular disease causing stroke, characterized by fibrocellular intimal thickening of cerebral arteries.
- Vascular smooth muscle cells (VSMCs) are key components of cerebral arteries, but their characteristics in MMD remain poorly understood.
- Induced pluripotent stem cell (iPSC) technology offers a novel approach to study MMD-associated VSMCs.
Purpose of the Study:
- To characterize VSMCs derived from MMD patient iPSCs.
- To compare the biological functions and transcriptome profiles of MMD-VSMCs with those from healthy controls (HC).
- To investigate the comparative transcriptome features between VSMCs and endothelial cells (ECs) in MMD.
Main Methods:
- Differentiated VSMCs from neural crest stem cells (NCSCs) using peripheral blood mononuclear cell-derived iPSCs from MMD patients and HC subjects.
- Compared biological characteristics (marker expression, proliferation, migration, contractility) and transcriptome profiles of VSMCs.
- Compared transcriptome profiles of iPSC-derived VSMCs and ECs.
Main Results:
- iPSC-derived VSMCs from MMD patients and HC subjects exhibited similar marker expression, proliferation, migration, and contractile abilities.
- Transcriptome analysis revealed similar gene expression profiles in MMD-VSMCs and HC-VSMCs, with only six differentially expressed genes (DEGs).
- Endothelial cells (ECs) from MMD patients showed a distinct transcriptome profile with 120 DEGs compared to HC-ECs, and the Wnt-signaling pathway was significant in VSMCs.
Conclusions:
- This study successfully established VSMCs from MMD patient iPSCs, demonstrating comparable biological functions and transcriptome profiles to HC-VSMCs under standard culture conditions.
- Distinct transcriptome profiles in MMD-ECs compared to MMD-VSMCs suggest that endothelial cells may predominantly drive MMD pathology.
- The iPSC-derived VSMC model provides a valuable tool for further research into diagnostic and therapeutic targets for MMD.
