Single-Cell Multimodal Profiling of Atherosclerosis Identifies CD200 as a Cell Surface Lineage Marker of Vascular Smooth Muscle Cells and Their Derived Cells.
Single-Cell Multimodal Profiling of Atherosclerosis Identifies CD200 as a Cell Surface Lineage Marker of Vascular Smooth Muscle Cells and Their Derived Cells.
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动脉粥样硬化的单细胞多模式分析将 CD200 鉴定为血管平滑肌细胞及其衍生细胞的细胞表面谱系标记。
DOI：
10.1161/circulationaha.123.067092
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发表时间：
2024
期刊：
影响因子：
37.8
通讯作者：
Reilly,MuredachP
中科院分区：
文献类型：
--
作者：
Bashore,AlexanderC;Chung,Allen;Ibikunle,Chinyere;Yan,Hanying;Xue,Chenyi;Li,Mingyao;Bauer,RobertC;Reilly,MuredachP
动脉粥样硬化性心血管疾病是全球主要的死亡原因。血管平滑肌细胞(VSMCs)在动脉粥样硬化中起核心作用，因为它们具有将表型转化为保护性或有害状态的能力。然而，由于缺乏明确的VSMC表面标记，识别和示踪VSMC及其后代的能力受到限制。1对血管平滑肌细胞的研究必须使用直线赛跑的小鼠模型，这种模型既耗时又具有挑战性，而且在人类身上是不可行的。在这里，我们使用转录和表位序列的细胞索引(CITE-SEQ)来表征小鼠动脉粥样硬化中119种细胞表面蛋白的表型。我们还在人类动脉粥样硬化中进行了CITE-SEQ，发现CD200高表达，即使在表型调节的情况下也对VSMCs具有特异性。小鼠CITE-SEQ数据可在国家生物技术信息中心基因表达总集(数据库登录号GSE246779)中获得。所有其他数据可根据要求从相应的作者处获得。哥伦比亚大学机构动物保护和使用委员会批准了所有程序(批准AABQ5576)，所有程序都符合美国国立卫生研究院的指导方针。哥伦比亚大学机构审查委员会批准了IRB AAAJ2765和AAAR6796下的所有人类方案，并获得了所有个人的书面知情同意。为了表征动脉粥样硬化不同阶段的细胞类型，我们使用了雄性LDLR−/−，ROSA26LSL−ZsGreen1/+，MYH11-CreerT2小鼠，在他莫昔芬治疗后永久诱导VSMC中ZsGreen1的表达。在西方饮食喂养的4个不同持续时间(0、8、16和26周)，ZsGreen1+和ZsGreen1−细胞被提交进行CITE-SEQ分析(图[A])。共鉴定出13个细胞团，包括巨噬细胞、T细胞、内皮细胞、血管平滑肌细胞和成纤维细胞。在ZsGreen1+的基础上，VSMC来源的细胞包括VSMC 1、VSMC 2和调制的VSMC群体(图[B])。我们的分析发现CD200是VSMCs和VSMC来源的细胞的高表达和特异性标记(图[C]和[D])。在成纤维细胞和内皮细胞上的表达也明显降低和特异性降低。此外，CD200的表达在动脉粥样硬化的不同持续时间内保持一致，直至26周(图[E])。为了验证我们的CITE-SEQ发现，我们开发了一个流式细胞仪小组来确定CD200是否可以区分血管平滑肌细胞和成纤维细胞，并使用这个小组来分析来自ZsGreen1+/−小鼠的细胞(图[F])。排除CD45+白细胞和CD31+内皮细胞，剩下的细胞包括VSMCs和成纤维细胞。我们检测了CD200的表达，鉴定了CD200+和CD200−细胞的两个群体。然后，我们检查了每个人口的比例
Atherosclerotic cardiovascular disease is the leading cause of death worldwide. Vascular smooth muscle cells (VSMCs) play a central role in atherosclerosis because of their ability to transition phenotypically into either a protective or harmful state. However, the ability to identify and trace VSMCs and their progeny is limited because of a lack of well-defined VSMC surface markers. 1 Investigations into VSMCs must use lineagetracing mouse models, which are time-consuming, challenging to generate, and not feasible in humans. Here, we used cellular indexing of transcriptomes and epitopes sequencing (CITE-seq) to characterize 119 cell surface proteins phenotypically in mouse atherosclerosis. We also performed CITE-seq in human atherosclerosis and found that CD200 is highly expressed and specific for VSMCs that persists even with phenotypic modulation. Mouse CITE-seq data are available in the National Center for Biotechnology Information Gene Expression Omnibus (database accession number GSE246779). All other data are available from the corresponding author upon request. The Institutional Animal Care and Use Committee of Columbia University approved all procedures (approval AABQ5576), and all procedures were in accordance with National Institutes of Health guidelines. The Institutional Review Board of Columbia University approved all human protocols under IRB AAAJ2765 and AAAR6796, with written informed consent obtained from all individuals.To characterize cell types at different stages of atherosclerosis, we used male LDLr−/−, ROSA26LSL− ZsGreen1/+, Myh11-CreERT2 mice, which permanently induces ZsGreen1 expression in VSMCs after tamoxifen administration. At 4 different durations of Western diet feeding (0, 8, 16, and 26 weeks), ZsGreen1+ and ZsGreen1− cells were submitted for CITE-seq profiling (Figure [A]). A total of 13 cell clusters were identified, including macrophages, T cells, endothelial cells, VSMCs, and fibroblasts. On the basis of ZsGreen1+, VSMC-derived cells comprised VSMC 1, VSMC 2, and modulated VSMC populations (Figure [B]). Our analysis identified CD200 as a highly expressed and specific marker of VSMCs and VSMC-derived cells (Figure [C] and [D]). There was also a marked lower and less specific expression on fibroblast and endothelial cells. In addition, the expression of CD200 was consistent and maintained throughout different durations of atherosclerosis up to and including 26 weeks (Figure [E]). To validate our CITE-seq findings, we developed a flow cytometry panel to determine whether CD200 can discriminate between VSMC and fibroblasts, and used this panel to analyze cells from the ZsGreen1+/− mice (Figure [F]). CD45+ leukocytes and CD31+ endothelial cells were excluded, so that the remaining cells included VSMCs and fibroblasts. We examined the expression of CD200, identifying 2 populations of CD200+ and CD200− cells. Then, we inspected the proportion of each population