Taxonomy of fibroblasts and progenitors in the synovial joint at single-cell resolution.
Taxonomy of fibroblasts and progenitors in the synovial joint at single-cell resolution.
复制标题
DOI：
10.1136/ard-2021-221682
复制
发表时间：
2023-03
影响因子：
27.4
通讯作者：
中科院分区：
文献类型：
--
作者：
滑膜中的成纤维细胞包括内层中的成纤维样滑膜细胞 (FLS) 和下层中的 Thy1+ 结缔组织成纤维细胞。我们的目的是研究它们的发育起源以及与成年祖细胞的关系。为了区分源自胚胎关节间带的 Gdf5 谱系细胞和其他表达 Pdgfrα 的成纤维细胞和祖细胞，使用成年 Gdf5-Cre;Tom;Pdgfrα-H2BGFP 小鼠并诱导软骨损伤以激活祖细胞。从膝盖中分离细胞，根据发育起源通过荧光激活细胞分选对成纤维细胞和祖细胞进行分选，并通过单细胞 RNA 测序进行分析。使用流式细胞术和免疫组织化学进行验证。使用 Gdf5-Cre;Confetti 小鼠进行克隆谱系作图。在稳定状态下，Thy1+ 亚衬成纤维细胞具有混合个体发育。相比之下，Thy1-Prg4+ 衬里成纤维细胞主要源自胚胎关节间带，并包括与分子定义的 FLS 不同的表达 Prg4 的祖细胞。克隆谱系追踪揭示了 Gdf5 谱系成纤维细胞在衬层和亚衬层之间的区室化。损伤后，表达 Prg4 的祖细胞增殖和分化导致内膜增生，同时额外招募非 Gdf5 谱系细胞进入 FLS。与此一致的是，第二群增殖细胞富集在下层血管附近，提供了活化的多能细胞，预计会产生 Thy1+ 成纤维细胞，并进入 FLS 分化轨迹。调节成纤维细胞分化轨迹的转录程序被发现，确定 Sox5 和 Foxo1 是小鼠和人类的关键 FLS 转录因子。我们的研究结果绘制了健康和受伤膝盖中小鼠滑膜成纤维细胞和祖细胞的细胞图谱，并为控制成人滑膜关节的组织和维护的细胞和分子原理提供了新的见解。
Fibroblasts in synovium include fibroblast-like synoviocytes (FLS) in the lining and Thy1+ connective-tissue fibroblasts in the sublining. We aimed to investigate their developmental origin and relationship with adult progenitors. To discriminate between Gdf5-lineage cells deriving from the embryonic joint interzone and other Pdgfrα-expressing fibroblasts and progenitors, adult Gdf5-Cre;Tom;Pdgfrα-H2BGFP mice were used and cartilage injury was induced to activate progenitors. Cells were isolated from knees, fibroblasts and progenitors were sorted by fluorescence-activated cell-sorting based on developmental origin, and analysed by single-cell RNA-sequencing. Flow cytometry and immunohistochemistry were used for validation. Clonal-lineage mapping was performed using Gdf5-Cre;Confetti mice. In steady state, Thy1+ sublining fibroblasts were of mixed ontogeny. In contrast, Thy1-Prg4+ lining fibroblasts predominantly derived from the embryonic joint interzone and included Prg4-expressing progenitors distinct from molecularly defined FLS. Clonal-lineage tracing revealed compartmentalisation of Gdf5-lineage fibroblasts between lining and sublining. Following injury, lining hyperplasia resulted from proliferation and differentiation of Prg4-expressing progenitors, with additional recruitment of non-Gdf5-lineage cells, into FLS. Consistent with this, a second population of proliferating cells, enriched near blood vessels in the sublining, supplied activated multipotent cells predicted to give rise to Thy1+ fibroblasts, and to feed into the FLS differentiation trajectory. Transcriptional programmes regulating fibroblast differentiation trajectories were uncovered, identifying Sox5 and Foxo1 as key FLS transcription factors in mice and humans. Our findings blueprint a cell atlas of mouse synovial fibroblasts and progenitors in healthy and injured knees, and provide novel insights into the cellular and molecular principles governing the organisation and maintenance of adult synovial joints.
登录
查看更多内容
影响因子：
4.6
作者：
Chou, Ching-Heng;Jain, Vaibhav;Kraus, Virginia B.
通讯作者：
Kraus, Virginia B.
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
3.9
作者：
SCHUMACHER, BL;BLOCK, JA;KUETTNER, KE
通讯作者：
KUETTNER, KE
影响因子：
64.5
作者：
Snippert, Hugo J.;van der Flier, Laurens G.;Clevers, Hans
通讯作者：
Clevers, Hans
影响因子：
4.8
作者：
Li, Lei;Newton, Phillip T.;Chagin, Andrei S.
通讯作者：
Chagin, Andrei S.