成纤维细胞异质性将炎症微环境与慢性鼻窦炎中的组织重塑联系起来。

文献信息

中文摘要

背景： 慢性鼻窦炎（CRS）是一种异质性炎症性疾病，具有不同的内型和组织重塑。成纤维细胞参与细胞外基质重塑和免疫调节，但其在CRS中的异质性仍不清楚。
方法： 将来自21例CRS和健康对照样本的公共单细胞转录组数据与来自鼻息肉的原代成纤维细胞的批量RNA测序数据整合，这些细胞分别用磷酸盐缓冲液（PBS）、干扰素（IFN）-γ、白细胞介素（IL）-4或IL-17A刺激。分析了成纤维细胞亚群、拟时序轨迹以及成纤维细胞与免疫细胞之间推断的通讯。将细胞因子诱导的特征投射到成纤维细胞图谱上。使用COMPASS队列（NCT05598424）的一部分纵向RNA-seq数据，在糖皮质激素治疗期间，跨V1（基线）、V2（鼻用皮质类固醇治疗4周后）和V3（随后全身糖皮质激素治疗后）验证预定义的候选标记基因，包括C8亚群的IL-4相关标记和C5/C10亚群的IL-17A相关标记。使用线性混合效应模型评估时间表达变化，并进行V1-V2和V2-V3比较。
结果： 在CRS亚型中鉴定出12个转录上不同的成纤维细胞亚群（C0-C11）。C8成纤维细胞在嗜酸性CRSwNP中富集，并与2型炎症相关基因相关，包括CCL26和POSTN，而C5和C10与非嗜酸性CRSwNP和IL-17A相关炎症特征相关。拟时序分析提示从C4分别向C8和C10的两条轨迹。CellChat揭示了成纤维细胞与免疫细胞亚群之间的双向通讯。细胞因子刺激诱导了不同的转录反应：IFN-γ主要诱导抗原呈递和干扰素反应；IL-4诱导收缩和细胞骨架相关特征；IL-17A激活IL-17、核因子κB和肿瘤坏死因子信号通路。特征投射将IL-4相关特征主要与C8成纤维细胞联系起来，将IL-17A相关特征主要与C5和C10成纤维细胞联系起来。对糖皮质激素治疗患者的纵向分析发现了一个稳定的C8相关转录特征，其特征是在V1-V3时间点CCL26、POSTN、CISH和CCL13持续下调。
结论： CRS中的成纤维细胞表现出显著的异质性，并可能响应不同的炎症刺激而参与组织重塑和免疫调节。IL-4相关的C8状态和IL-17A相关的C5/C10状态可能代表与不同CRS炎症微环境相关的不同成纤维细胞状态。C8相关转录模块在糖皮质激素治疗下显示出稳定且持续的抑制。

英文摘要

BACKGROUND: Chronic rhinosinusitis (CRS) is a heterogeneous inflammatory disorder with distinct endotypes and tissue remodeling. Fibroblasts contribute to extracellular matrix remodeling and immune regulation, but their heterogeneity in CRS remains unclear.
METHODS: Public single-cell transcriptomic data from 21 CRS and healthy control samples were integrated with bulk RNA sequencing data from primary fibroblasts derived from nasal polyps stimulated with phosphate buffered saline (PBS), interferon (IFN)-γ, interleukin (IL)-4, or IL-17A. Fibroblast subsets, pseudotime trajectories, and inferred communication between fibroblasts and immune cells were analyzed. Cytokine-induced signatures were projected onto the fibroblast atlas. A subset of longitudinal RNA-seq data from the COMPASS cohort (NCT05598424) was used to validate predefined candidate marker genes, including IL-4-associated markers of the C8 subset and IL-17A-associated markers of the C5/C10 subsets, during glucocorticoid treatment across V1 (baseline), V2 (after 4 weeks of intranasal corticosteroid treatment), and V3 (after subsequent systemic glucocorticoid therapy). Temporal expression changes were assessed using linear mixed-effects models with V1-V2 and V2-V3 comparisons.
RESULTS: Twelve transcriptionally distinct fibroblast subsets (C0-C11) were identified across CRS subtypes. C8 fibroblasts were enriched in eosinophilic CRSwNP and associated with type 2 inflammation related genes, including CCL26 and POSTN, whereas C5 and C10 were associated with non-eosinophilic CRSwNP and IL-17A related inflammatory features. Pseudotime analysis suggested two trajectories from C4 toward C8 and C10 respectively. CellChat revealed bidirectional communication between fibroblasts and immune cell subsets. Cytokine stimulation induced distinct transcriptional responses: IFN-γ mainly induced antigen presentation and interferon response; IL-4 induced contraction and cytoskeleton related features; and IL-17A activated IL-17, nuclear factor κB, and tumor necrosis factor signaling pathways. Signature projection linked IL-4 related features mainly to C8 fibroblasts and IL-17A related features mainly to C5 and C10 fibroblasts. Longitudinal analysis of glucocorticoid-treated patients identified a stable C8-associated transcriptional signature characterized by sustained downregulation of CCL26, POSTN, CISH, and CCL13 across V1-V3 time points.
CONCLUSIONS: Fibroblasts in CRS exhibit marked heterogeneity and may contribute to tissue remodeling and immune regulation in response to distinct inflammatory stimuli. The IL-4-associated C8 state and IL-17A-associated C5/C10 states may represent distinct fibroblast states associated with different CRS inflammatory microenvironments. C8-associated transcriptional module shows stable and sustained suppression under glucocorticoid treatment.