The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST), Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Department of Oral and Maxillofacial Head Neck Surgery, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Oral Dis. 2023 Nov;29(8):3420-3432. doi: 10.1111/odi.14293. Epub 2022 Jul 10.
Various types of cells comprising a complex and diverse cell population are required for the biological activities of odontogenic keratocyst (OKC). Immune and non-immune cells collaborate via cytokine- or chemokine-mediated communication and direct cell-cell interactions. This study aimed to characterize the immune ecosystem and understand the potential chemotactic role of OKC fibroblasts in immune cell migration.
Mass cytometry of 41 markers was employed for the classification of OKC cells from six OKC samples. Immunofluorescence staining and single-cell RNA sequencing (GSE176351) were used for the detection of fibroblast subpopulations. Enzyme-linked immunosorbent assay and immunofluorescence staining were employed for chemokine detection in hypoxia- and/or HIF-1α inhibitor-treated OKC fibroblasts and tissues. Chemotaxis assay was employed to determine the chemotactic effect of fibroblasts via co-culture with peripheral blood mononuclear cells. A cell communication network was constructed based on the single-cell RNA sequencing data.
The characterization of the immune cell types of OKC evidenced the enrichment of macrophages, neutrophils and B cells. The majority (41.5%) of fibroblast subsets consisted of chemokine ligand-enriched myofibroblasts. The activation of the HIF-1α signaling pathway in fibroblasts was associated with chemokine release. The chemokines released by OKC fibroblasts remarkably promoted the migration of peripheral blood mononuclear cells in the co-culture system. Close interactions between myofibroblasts and immune cells were validated by cell-cell interaction analysis. Increased RANKL expression was detected in OKC fibroblasts in the co-culture system with peripheral blood mononuclear cells.
Our results provided deep insights into the immune ecosystem and highlighted the potential chemotactic effects of chemokine-enriched myofibroblasts within OKCs. The close interaction between immune cells and fibroblasts demonstrated in this study may be responsible for the osteoclastogenic effects of OKC fibroblasts.
牙源性角化囊性瘤（OKC）的生物学活性需要多种类型的细胞构成复杂多样的细胞群体。免疫细胞和非免疫细胞通过细胞因子或趋化因子介导的通讯以及直接的细胞-细胞相互作用进行协作。本研究旨在描绘 OKC 的免疫生态系统，并探讨 OKC 成纤维细胞在免疫细胞迁移中的潜在趋化作用。
采用 41 种标志物的液质联用技术对来自 6 个 OKC 样本的 OKC 细胞进行分类。免疫荧光染色和单细胞 RNA 测序（GSE176351）用于检测成纤维细胞亚群。酶联免疫吸附试验和免疫荧光染色用于检测缺氧和/或 HIF-1α 抑制剂处理后的 OKC 成纤维细胞和组织中的趋化因子。通过与外周血单个核细胞共培养来进行趋化实验，以确定成纤维细胞的趋化作用。基于单细胞 RNA 测序数据构建细胞通讯网络。
OKC 免疫细胞类型的特征表明巨噬细胞、中性粒细胞和 B 细胞明显富集。大多数（41.5%）成纤维细胞亚群由富含趋化因子配体的肌成纤维细胞组成。成纤维细胞中 HIF-1α 信号通路的激活与趋化因子的释放有关。OKC 成纤维细胞释放的趋化因子在共培养系统中显著促进外周血单个核细胞的迁移。通过细胞-细胞相互作用分析验证了肌成纤维细胞与免疫细胞之间的紧密相互作用。在与外周血单个核细胞共培养的系统中检测到 OKC 成纤维细胞中 RANKL 表达增加。
本研究结果深入了解了免疫生态系统，并强调了 OKC 中富含趋化因子的肌成纤维细胞的潜在趋化作用。本研究中观察到的免疫细胞和成纤维细胞之间的紧密相互作用可能是 OKC 成纤维细胞破骨作用的原因。
