- 作者: Li Hao, Xiaolong Ya, Jiaye Wu, Chuming Tao, Ruochen Ma, Zhiyao Zheng, Siqi Mou, Yiming Ling, Yingxi Yang, Jiguang Wang, Yan Zhang, Qing Lin & Jizong Zhao
- 作者服務機構: 1.China National Clinical Research Center for Neurological Diseases, Beijing, 100070, China 2.Department of Chemical and Biological Engineering, Division of Life Science, State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Hong Kong SAR, China 3.Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Hong Kong, China 4.Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, 100070, China 5.Division of Cell, Developmental and Integrative Biology, School of Medicine, South China University of Technology, Guangzhou, 510006, China 6.HKUST Shenzhen-Hong Kong Collaborative Innovation Research Institute, Futian, Shenzhen, China 7.Hong Kong Center for Neurodegenerative Diseases, InnoHK, HKSAR, China 8.Joint Laboratory of School of Pharmacy, Capital Medical University and National Clinical Research Center for Nervous System Diseases, Beijing, China 9.Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, China 10.University of Chinese Academy of Sciences, Beijing, 100049, China
- 中文摘要:
- 英文摘要:
Background
The fusiform aneurysm is a nonsaccular dilatation affecting the entire vessel wall over a short distance. Although PDGFRB somatic variants have been identified in fusiform intracranial aneurysms, the molecular and cellular mechanisms driving fusiform intracranial aneurysms due to PDGFRB somatic variants remain poorly understood.
Methods
In this study, single-cell sequencing and immunofluorescence were employed to investigate the phenotypic changes in smooth muscle cells within fusiform intracranial aneurysms. Whole-exome sequencing revealed the presence of PDGFRB gene mutations in fusiform intracranial aneurysms. Subsequent immunoprecipitation experiments further explored the functional alterations of these mutated PDGFRB proteins. For the common c.1684 mutation site of PDGFRβ, we established mutant smooth muscle cell lines and zebrafish models. These models allowed us to simulate the effects of PDGFRB mutations. We explored the major downstream cellular pathways affected by PDGFRBY562D mutations and evaluated the potential therapeutic effects of Ruxolitinib.
Results
Single-cell sequencing of two fusiform intracranial aneurysms sample revealed downregulated smooth muscle cell markers and overexpression of inflammation-related markers in vascular smooth muscle cells, which was validated by immunofluorescence staining, indicating smooth muscle cell phenotype modulation is involved in fusiform aneurysm. Whole-exome sequencing was performed on seven intracranial aneurysms (six fusiform and one saccular) and PDGFRB somatic mutations were detected in four fusiform aneurysms. Laser microdissection and Sanger sequencing results indicated that the PDGFRB mutations were present in smooth muscle layer. For the c.1684 (chr5: 149505131) site mutation reported many times, further cell experiments showed that PDGFRBY562D mutations promoted inflammatory-related vascular smooth muscle cell phenotype and JAK-STAT pathway played a crucial role in the process. Notably, transfection of PDGFRBY562D in zebrafish embryos resulted in cerebral vascular anomalies. Ruxolitinib, the JAK inhibitor, could reversed the smooth muscle cells phenotype modulation in vitro and inhibit the vascular anomalies in zebrafish induced by PDGFRB mutation.
Conclusion
Our findings suggested that PDGFRB somatic variants played a role in regulating smooth muscle cells phenotype modulation in fusiform aneurysms and offered a potential therapeutic option for fusiform aneurysms. - 中文關鍵字:
- 英文關鍵字: PDGFRB variants, Phenotype modulation, Intracranial fusiform aneurysm, Smooth muscle cell