背景:脑海绵状畸形(CCM)是一种低流量,容易出血的血管疾病,可导致脑出血,癫痫发作和神经功能缺损。其遗传方式包括偶发性或常染色体显性遗传,外显率不完全,即散发性CCM(SCCM)和家族性CCM。SCCM在家庭中的特点是单一病变和单一情感。在CCM患者中,尤其是SCCM,相应表型和病理特征或候选基因的发病机制尚未完全阐明。
方法:这里,我们在SCCM和对照组患者中进行了深入的单细胞RNA测序(scRNA-Seq)和转座酶可接近染色质测序(ATAC-Seq)的批量测定.使用qPCR和RNA原位杂交(RNAFISH)技术对感兴趣的基因进行进一步验证,以提供进一步的图谱和SCCM生成过程的证据。
结果:我们在SCCM和对照血管中鉴定了六种细胞类型,发现NEK1,RNPC3,FBRSL1,IQGAP2,MCUB,AP3B1、ESCO1、MYO9B和PVT1在SCCM组织中表达上调。在六种细胞类型中,我们发现,与对照条件相比,PVT1在SCCM样品的内皮细胞簇中显示出一个上升的峰值,该峰值跟随伪时间轴,同时在SCCM样品的平滑肌细胞簇中显示出增加的趋势。进一步的实验表明,与对照船只相比,PVT1在SCCM样品中表现出显著升高的表达。
结论:在SCCM条件下,我们发现在从控制到病变的发展过程中,PVT1在内皮细胞中呈现上升峰,同时在平滑肌细胞中呈现上升趋势。总的来说,NEK1,RNPC3,FBRSL1,IQGAP2,MCUB的表达显着升高,与对照样品相比,SCCM样品中的AP3B1、ESCO1、MYO9B和PVT1。
BACKGROUND: Cerebral cavernous malformation (
CCM) is a low-flow, bleeding-prone vascular disease that can cause cerebral hemorrhage, seizure and neurological deficits. Its inheritance mode includes sporadic or autosomal dominant inheritance with incomplete penetrance, namely sporadic
CCM (SCCM) and familial
CCM. SCCM is featured by single lesion and single affection in a family. Among
CCM patients especially SCCM, the pathogenesis of the corresponding phenotypes and pathological features or candidate genes have not been fully elucidated yet.
METHODS: Here, we performed in-depth single-cell RNA sequencing (scRNA-Seq) and bulk assay for transposase-accessible chromatin sequencing (ATAC-Seq) in SCCM and control patients. Further validation was conducted for the gene of interest using qPCR and RNA in situ hybridization (RNA FISH) techniques to provide further atlas and evidence for SCCM generative process.
RESULTS: We identified six cell types in the SCCM and control vessels and found that the expression of NEK1, RNPC3, FBRSL1, IQGAP2, MCUB, AP3B1, ESCO1, MYO9B and PVT1 were up-regulated in SCCM tissues. Among the six cell types, we found that compared with control conditions, PVT1 showed a rising peak which followed the pseudo-time axis in endothelial cell clusters of SCCM samples, while showed an increasing trend in smooth muscle cell clusters of SCCM samples. Further experiments indicated that, compared with the control vessels, PVT1 exhibited significantly elevated expression in SCCM samples.
CONCLUSIONS: In SCCM conditions, We found that in the process of development from control to lesion conditions, PVT1 showed a rising peak in endothelial cells and showed an increasing trend in smooth muscle cells at the same time. Overall, there was a significantly elevated expression of NEK1, RNPC3, FBRSL1, IQGAP2, MCUB, AP3B1, ESCO1, MYO9B and PVT1 in SCCM specimens compared to control samples.