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Experimental Approach to Moyamoya Angiopathy: Insights into Vascular Cell Crosstalk
Gemma Gorla1, Antonella Potenza1,2, Tatiana Carrozzini1
1Laboratory of Neurobiology and Cerebrovascular Unit, Fondazione IRCCS Istituto Neurologico Carlo Besta, 20133 Milan, Italy.
Background:
The pathophysiological mechanisms of Moyamoya angiopathy (MA) are still largely unknown, although a dysfunctional vasculogenesis has been hypothesized to contribute to it. The association between this rare cerebrovascular condition and variants of Ring Finger Protein 213 (RNF213) strengthens the role of genetic factors in MA pathogenesis.
Methods:
To investigate the molecular mechanisms of MA, we carried out RNA interference (RNAi) targeting RNF213 in human endothelial cells (ECs) and vascular smooth muscle cells (VSMCs). The combined effect of RNAi and/or hypoxia on expression of key angiogenic factors was analyzed through qRT-PCR and Western blot. Functional assays were performed to characterize the impact of RNAi on vasculogenesis. Gene-expression arrays were performed on vessel walls of MA patients and controls.
Results:
RNF213-RNAi impaired angiogenic capability in ECs, whereas the simultaneous silencing of RNF213 and its phosphatase PTP1B restored angiogenesis function in ECs but worsened it in VSMCs. Angiogenic factor expression appeared to be modulated in ECs by the combined effects of RNAi and/or hypoxia, and in pathological vessels of MA patients as compared with controls.
Conclusions:
Our findings contribute to associating the relevance of RNF213 in MA cellular models and highlight the importance of EC-VSMC crosstalk for vascular integrity. Additionally, the study could lay the foundations for improving experimental models of MA pathophysiology.
Insights
Ring Finger Protein 213 (RNF213) plays a key role in Moyamoya angiopathy (MA) pathogenesis. Its dysfunction impairs angiogenesis, but interactions with PTP1B and cell types (endothelial cells and vascular smooth muscle cells) influence vascular integrity.
Area of Science:
- Molecular Biology
- Genetics
- Cerebrovascular Diseases
Background:
- Moyamoya angiopathy (MA) pathophysiology remains unclear, with dysfunctional vasculogenesis implicated.
- Genetic factors, particularly Ring Finger Protein 213 (RNF213) variants, are strongly associated with MA.
Purpose of the Study:
- To investigate the molecular mechanisms underlying MA.
- To explore the role of RNF213 in endothelial cells (ECs) and vascular smooth muscle cells (VSMCs).
Main Methods:
- RNA interference (RNAi) targeting RNF213 in ECs and VSMCs.
- Analysis of angiogenic factor expression via qRT-PCR and Western blot.
- Gene-expression arrays on patient and control vessel walls.
Main Results:
- RNF213-RNAi reduced angiogenic capacity in ECs.
- Co-silencing RNF213 and PTP1B restored EC angiogenesis but impaired VSMC function.
- Angiogenic factor expression was modulated by RNF213, hypoxia, and cell type.
Conclusions:
- RNF213 is relevant in MA cellular models.
- Endothelial cell-vascular smooth muscle cell crosstalk is crucial for vascular integrity in MA.
- Findings may improve MA pathophysiology experimental models.
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