Ibikunle Chinyere O, Xue Chenyi, Kim Eunyoung, Yan Hanying, Coronel Johana, Zhu Lucie Y, Cui Jian, Chung Allen, Bauer Robert C, Sachs Nadja, Maegdefessel Lars, Li Mingyao, Tall Alan R, Bashore Alexander C, Reilly Muredach P
Division of Cardiology, Department of Medicine, Vagelos College of Physicians and Surgeons, New York, NY.
Cardiometabolic Genomics Program, Division of Cardiology, Department of Medicine Columbia University Irving Medical Center, New York, NY.
bioRxiv. 2025 May 23:2025.05.20.655228. doi: 10.1101/2025.05.20.655228.
Vascular smooth muscle cells (VSMCs) play a central role in atherosclerosis by undergoing phenotypic modulation from a quiescent, contractile state to a range of synthetic phenotypes, including fibroblast-like, macrophage-like, and lipid-laden foam cell-like states. However, a comprehensive multimodal characterization and understanding of the transcriptional programs driving these transitions remain incomplete.
To comprehensively define the phenotypic diversity of VSMCs during atherosclerosis progression, we performed in-depth profiling using cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) and bulk RNA sequencing in a VSMC lineage-tracing atherosclerotic mouse model. Insights from these datasets guided the design of targeted in vitro experiments to investigate candidate regulatory mechanisms.
Single-cell multi-omics revealed extensive cellular heterogeneity within atherosclerotic plaques, including a rare population of VSMC-derived macrophage-like cells, whose presence was confirmed by histological analysis. These studies also identified a substantial population of VSMC-derived foam cells, comprising approximately 70% of all foam cells in the lesions. These cells exhibited activation of gene programs associated with lipid metabolism, proliferation, and tumor-like features. The transcription factor Bhlhe40 emerged as a key regulator of this phenotypic transition, with elevated expression in VSMC-derived foam cells during disease progression. Functional knockdown of Bhlhe40 suppressed VSMC phenotypic switching and foam cell characteristics, underscoring its potential role as a driver of VSMC modulation.
These findings advance our understanding of VSMC phenotypic modulation in atherosclerosis and highlight Bhlhe40 as a key regulator of this process. Elucidating the mechanisms governing VSMC plasticity may offer new therapeutic opportunities to reduce cardiovascular risk by targeting disease-driving cellular transitions.
血管平滑肌细胞（VSMCs）在动脉粥样硬化中起着核心作用，通过从静止的收缩状态转变为一系列合成表型，包括成纤维细胞样、巨噬细胞样和富含脂质的泡沫细胞样状态。然而，对驱动这些转变的转录程序进行全面的多模式表征和理解仍不完整。
为了全面定义动脉粥样硬化进展过程中VSMCs的表型多样性，我们在VSMC谱系追踪动脉粥样硬化小鼠模型中，使用转录组和表位测序的细胞索引（CITE-seq）和批量RNA测序进行了深入分析。这些数据集的见解指导了靶向体外实验的设计，以研究候选调控机制。
单细胞多组学揭示了动脉粥样硬化斑块内广泛的细胞异质性，包括罕见的VSMC衍生的巨噬细胞样细胞群体，其存在通过组织学分析得到证实。这些研究还确定了大量的VSMC衍生的泡沫细胞群体，约占病变中所有泡沫细胞的70%。这些细胞表现出与脂质代谢、增殖和肿瘤样特征相关的基因程序的激活。转录因子Bhlhe40成为这种表型转变的关键调节因子，在疾病进展过程中VSMC衍生的泡沫细胞中表达升高。Bhlhe40的功能敲低抑制了VSMC表型转换和泡沫细胞特征，强调了其作为VSMC调节驱动因素的潜在作用。
这些发现推进了我们对动脉粥样硬化中VSMC表型调节的理解，并突出了Bhlhe40作为这一过程的关键调节因子。阐明控制VSMC可塑性的机制可能为通过靶向疾病驱动的细胞转变来降低心血管风险提供新的治疗机会。
