体细胞表皮生长因子受体活化变异促进颅内纺锤形动脉瘤平滑肌细胞表型的改变。

IF 9 2区 医学 Q1 CELL BIOLOGY Journal of Biomedical Science Pub Date : 2024-05-13 DOI:10.1186/s12929-024-01040-7
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
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引用次数: 0

摘要

背景:纺锤形动脉瘤是一种在短距离内影响整个血管壁的非圆形扩张。虽然已在纺锤形颅内动脉瘤中发现了 PDGFRB 体细胞变异,但对 PDGFRB 体细胞变异导致纺锤形颅内动脉瘤的分子和细胞机制仍知之甚少:本研究采用单细胞测序和免疫荧光技术研究了纺锤形颅内动脉瘤内平滑肌细胞的表型变化。全外显子组测序显示,纺锤形颅内动脉瘤中存在 PDGFRB 基因突变。随后的免疫沉淀实验进一步探究了这些突变的 PDGFRB 蛋白的功能变化。针对 PDGFRβ 的常见 c.1684 突变位点,我们建立了突变平滑肌细胞系和斑马鱼模型。通过这些模型,我们可以模拟 PDGFRB 突变的影响。我们探索了受PDGFRBY562D突变影响的主要下游细胞通路,并评估了Ruxolitinib的潜在治疗效果:对两例纺锤形颅内动脉瘤样本进行单细胞测序发现,血管平滑肌细胞中平滑肌细胞标志物下调,炎症相关标志物过度表达,免疫荧光染色验证了这一点,表明平滑肌细胞表型调控参与了纺锤形动脉瘤的发生。对 7 个颅内动脉瘤(6 个纺锤形动脉瘤和 1 个囊状动脉瘤)进行了全外显子组测序,在 4 个纺锤形动脉瘤中检测到了 PDGFRB 体细胞突变。激光显微切割和 Sanger 测序结果表明,PDGFRB 突变存在于平滑肌层。对于多次报道的c.1684(chr5: 149505131)位点突变,进一步的细胞实验表明,PDGFRBY562D突变促进了炎症相关的血管平滑肌细胞表型,而JAK-STAT通路在这一过程中发挥了关键作用。值得注意的是,在斑马鱼胚胎中转染 PDGFRBY562D 会导致脑血管异常。JAK抑制剂Ruxolitinib可逆转体外平滑肌细胞表型的改变,并抑制PDGFRB突变诱导的斑马鱼血管异常:我们的研究结果表明,PDGFRB体细胞变异在纺锤形动脉瘤平滑肌细胞表型调控中发挥了作用,为纺锤形动脉瘤提供了一种潜在的治疗方案。
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Somatic PDGFRB activating variants promote smooth muscle cell phenotype modulation in intracranial fusiform aneurysm.

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.

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来源期刊
Journal of Biomedical Science
Journal of Biomedical Science 医学-医学:研究与实验
CiteScore
18.50
自引率
0.90%
发文量
95
审稿时长
1 months
期刊介绍: The Journal of Biomedical Science is an open access, peer-reviewed journal that focuses on fundamental and molecular aspects of basic medical sciences. It emphasizes molecular studies of biomedical problems and mechanisms. The National Science and Technology Council (NSTC), Taiwan supports the journal and covers the publication costs for accepted articles. The journal aims to provide an international platform for interdisciplinary discussions and contribute to the advancement of medicine. It benefits both readers and authors by accelerating the dissemination of research information and providing maximum access to scholarly communication. All articles published in the Journal of Biomedical Science are included in various databases such as Biological Abstracts, BIOSIS, CABI, CAS, Citebase, Current contents, DOAJ, Embase, EmBiology, and Global Health, among others.
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