综合生物信息学分析和实验验证揭示了昼夜节律相关基因在骨质疏松症和动脉粥样硬化之间的相互作用

摘要(中文译文)

骨质疏松症（OP）和动脉粥样硬化（AS）在老年人群中越来越普遍，并且经常共存，具有异常血管钙化等病理特征。然而，OP和AS之间相互作用的共同标记基因仍然知之甚少。本研究旨在鉴定导致OP和AS共存的共享标记基因和细胞机制。从基因表达综合数据库（GEO）中获得了与OP和AS相关的单细胞RNA测序（scRNA-seq）数据集和mRNA表达谱。使用R软件包Seurat、AUCell和hdWGCNA进行生物信息学分析，以鉴定昼夜节律相关基因（CRG）和相关细胞群。建立了自然衰老小鼠和0.25%腺嘌呤（AD）诱导的小鼠模型。进行了微计算机断层扫描（µCT）、免疫组织化学（IHC）、免疫荧光、Von Kossa染色和RT-qPCR，以实验验证鉴定的常见基因。在人类骨髓样本中，鉴定出9个细胞谱系，包括17个亚群，而在颈动脉样本中鉴定出6个细胞谱系和18个亚群。OP患者的骨髓基质细胞（BMSCs）和AS患者的血管平滑肌细胞（VSMCs）的昼夜节律活性显著增强。通过整合scRNA-seq和体RNA-seq分析，鉴定出69个常见基因，其中15个中心基因通过蛋白质相互作用（PPI）网络分析进一步筛选。实验验证表明，AEBP1在自然衰老和AD诱导的小鼠中显著上调，伴有PLIN1表达增加、骨钙素（OCN）表达减少和血管钙化增强。本研究揭示，OP中BMSCs和AS中VSMCs的昼夜节律相关调节在它们的共存中起着关键作用。此外，我们确定了共享的标记基因，特别是AEBP1，它们可能成为OP和as并发发展的潜在生物标志物和治疗靶点。

Abstract

Osteoporosis (OP) and atherosclerosis (AS) are increasingly prevalent in aging populations and frequently coexist, sharing pathological features such as abnormal vascular calcification. However, the common marker genes underlying the crosstalk between OP and AS remain poorly understood. This study aimed to identify shared marker genes and cellular mechanisms contributing to the coexistence of OP and AS. Single-cell RNA sequencing (scRNA-seq) dataset and mRNA expression profiles related to OP and AS were obtained from the Gene Expression Omnibus (GEO) database. Bioinformatics analyses were performed using the R packages Seurat, AUCell, and hdWGCNA to identify circadian rhythm-related genes (CRGs) and associated cell populations. Naturally aged mice and a 0.25% adenine (AD)-induced mouse model were established. Micro-computed tomography (µCT), immunohistochemistry (IHC), immunofluorescence, Von Kossa staining, and RT-qPCR were conducted to experimentally validate the identified common genes. In human bone marrow samples, nine cell lineages comprising 17 subpopulations were identified, while six cell lineages comprising 18 subpopulations were identified in carotid artery samples. Circadian rhythm activity was significantly enriched in bone marrow stromal cells (BMSCs) in OP and vascular smooth muscle cells (VSMCs) in AS. By integrating scRNA-seq and bulk RNA-seq analyses, 69 common genes were identified, of which 15 hub genes were further selected through protein-protein interaction (PPI) network analysis. Experimental validation demonstrated that AEBP1 was markedly upregulated in both naturally aged and AD-induced mice, accompanied by increased PLIN1 expression, reduced osteocalcin (OCN) expression, and enhanced vascular calcification. This study reveals that circadian rhythm-related regulation of BMSCs in OP and VSMCs in AS plays a critical role in their coexistence. Furthermore, we identify shared marker genes, particularly AEBP1, which may serve as potential biomarkers and therapeutic targets for the concurrent development of OP and AS.

如何引用

Mammalian genome : official journal of the International Mammalian Genome Society 的其他文献

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