Multi-omic single cell analysis resolves novel stromal cell populations in healthy and diseased human tendon
Multi-omic single cell analysis resolves novel stromal cell populations in healthy and diseased human tendon
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DOI：
10.1038/s41598-020-70786-5
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发表时间：
2020-09-03
影响因子：
4.6
通讯作者：
Carr, Andrew
中科院分区：
文献类型：
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作者：
Kendal, Adrian R.;Layton, Thomas;Carr, Andrew
肌腱病占初级保健咨询的30%以上，代表了活跃和日益老龄化的人口中日益增长的医疗保健挑战。识别参与肌腱病的关键细胞对于开发治疗方法以应对这一挑战至关重要。肌腱细胞是异质的，稀疏分布在致密的胶原蛋白基质，限制了以前的方法来研究细胞特性离体。我们应用了下一代CITE测序;将表面蛋白质组学与来自11个慢性肌腱病和8个健康人肌腱的>6400个单细胞的离体深入、无偏见的基因表达分析相结合。免疫组化证实了单细胞结果。我们第一次表明，人类肌腱窝藏至少五个不同的COL 1A 1/2表达肌腱细胞群，除了内皮细胞，T细胞和单核细胞。这些细胞包括表达微纤维相关基因的KRT 7/SCX+细胞、共表达高水平促炎标志物的PTX 3+细胞、APOD+纤维脂肪生成祖细胞、TPPP 3/PRG 4+软骨生成细胞和ITGA 7+平滑肌间充质细胞。表面蛋白质组学分析确定了这些亚类可以在未来分离和靶向的标记。慢性肌腱病与微原纤维相关肌腱细胞的促炎标记物PTX 3、CXCL 1、CXCL 6、CXCL 8和PDPN的表达增加相关。内皮细胞损伤增加了趋化因子和alarmin基因的表达，包括IL 33。
Tendinopathy accounts for over 30% of primary care consultations and represents a growing healthcare challenge in an active and increasingly ageing population. Recognising critical cells involved in tendinopathy is essential in developing therapeutics to meet this challenge. Tendon cells are heterogenous and sparsely distributed in a dense collagen matrix; limiting previous methods to investigate cell characteristics ex vivo. We applied next generation CITE-sequencing; combining surface proteomics with in-depth, unbiased gene expression analysis of>6400 single cells ex vivo from 11 chronically tendinopathic and 8 healthy human tendons. Immunohistochemistry validated the single cell findings. For the first time we show that human tendon harbours at least five distinct COL1A1/2 expressing tenocyte populations in addition to endothelial cells, T-cells, and monocytes. These consist of KRT7/SCX+ cells expressing microfibril associated genes, PTX3+ cells co-expressing high levels of pro-inflammatory markers, APOD+ fibro-adipogenic progenitors, TPPP3/PRG4+ chondrogenic cells, and ITGA7+ smooth muscle-mesenchymal cells. Surface proteomic analysis identified markers by which these sub-classes could be isolated and targeted in future. Chronic tendinopathy was associated with increased expression of pro-inflammatory markers PTX3, CXCL1, CXCL6, CXCL8, and PDPN by microfibril associated tenocytes. Diseased endothelium had increased expression of chemokine and alarmin genes including IL33.