Bashore Alexander C, Coronel Johana, Xue Chenyi, Zhu Lucie Y, Reilly Muredach P
Cardiovascular Research Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
Division of Cardiology, Department of Medicine, Vagelos College of Physicians and Surgeons, Columbia University, New York, New York, USA.
bioRxiv. 2025 Jan 28:2025.01.27.635067. doi: 10.1101/2025.01.27.635067.
Atherosclerosis involves complex interactions between lipids, immune cells, vascular smooth muscle cells (VSMCs), and fibroblasts within the arterial wall. While significant advances in single-cell technologies have shed light on the roles of immune cells and VSMCs in plaque development, fibroblasts remain underexplored, leaving critical gaps in understanding their contributions to disease progression and plaque stability. Comprehensive characterization of fibroblast phenotypes in atherosclerosis is essential to unravel their diverse functions and to distinguish between subsets that may play protective versus pathogenic roles in the disease process.
Here, we utilized CITE-seq (Cellular Indexing of Transcriptomes and Epitopes by Sequencing) to comprehensively profile fibroblast diversity in a mouse model of atherosclerosis. Mice were fed an atherogenic diet for 0, 8, 19, and 26 weeks, representing distinct stages of disease progression, enabling a detailed phenotypic characterization of fibroblasts throughout the course of atherosclerosis development.
We identified four distinct fibroblast subpopulations, including a myofibroblast population closely resembling VSMC-derived chondromyocytes. The proportions of these fibroblast subsets exhibited a modest decline as atherosclerosis progressed. Through multimodal analysis, we identified CD26 as a highly expressed and specific marker for one of these fibroblast subpopulations, distinguishing it from other subsets. Using a combination of flow cytometry and immunohistochemistry, we demonstrated that CD26 fibroblasts predominantly reside in the adventitia of healthy arteries. During atherosclerosis progression, these cells expand into the intima and primarily localize within the fibrous cap of the lesion.
Our multi-omic analysis highlights the phenotypic diversity and dynamic changes of fibroblasts during atherosclerosis progression. Among these, CD26+ fibroblasts emerge as a distinct subpopulation that expands within atherosclerotic lesions and may play a critical role in promoting plaque stability through their migration into the fibrous cap.
动脉粥样硬化涉及动脉壁内脂质、免疫细胞、血管平滑肌细胞（VSMC）和成纤维细胞之间的复杂相互作用。虽然单细胞技术取得了重大进展，揭示了免疫细胞和VSMC在斑块形成中的作用，但成纤维细胞仍未得到充分研究，在理解它们对疾病进展和斑块稳定性的贡献方面存在关键差距。全面表征动脉粥样硬化中成纤维细胞的表型对于阐明其多样功能以及区分在疾病过程中可能起保护作用与致病作用的亚群至关重要。
在此，我们利用CITE-seq（通过测序对转录组和表位进行细胞索引）全面分析动脉粥样硬化小鼠模型中成纤维细胞的多样性。给小鼠喂食致动脉粥样硬化饮食0、8、19和26周，代表疾病进展的不同阶段，从而能够在动脉粥样硬化发展过程中对成纤维细胞进行详细的表型表征。
我们鉴定出四个不同的成纤维细胞亚群，包括一个与VSMC衍生的软骨样细胞非常相似的肌成纤维细胞群体。随着动脉粥样硬化的进展，这些成纤维细胞亚群的比例略有下降。通过多模态分析，我们确定CD26是其中一个成纤维细胞亚群的高表达特异性标志物，将其与其他亚群区分开来。通过流式细胞术和免疫组织化学相结合的方法，我们证明CD26成纤维细胞主要存在于健康动脉的外膜中。在动脉粥样硬化进展过程中，这些细胞扩展到内膜并主要定位于病变的纤维帽内。
我们的多组学分析突出了动脉粥样硬化进展过程中成纤维细胞的表型多样性和动态变化。其中，CD26 +成纤维细胞作为一个独特的亚群出现，在动脉粥样硬化病变中扩张，并可能通过迁移到纤维帽中在促进斑块稳定性方面发挥关键作用。
