宫颈癌（CC）是全球女性第四大常见恶性肿瘤，也是发展中国家女性癌症相关死亡的主要原因。 CC 的早期症状通常不明显，通常在晚期才做出诊断，这导致临床预后不佳。近年来，大量研究表明肥大细胞（MCs）与肿瘤发生发展有着密切的关系。然而，当时对于MC在CC中所发挥的作用的研究还非常有限。因此，该研究对人CC细胞进行了单细胞多组学分析，旨在探讨MCs与CC肿瘤微环境相互作用的机制。目的是为CC的预防、诊断和治疗提供科学依据，以期改善患者的预后和生活质量。本研究通过ArrayExpress获取了10个CC肿瘤样本的单细胞RNA测序数据数据库。利用 Slingshot 和 AUCcell 来推断和评估 MC 亚群的分化轨迹和细胞可塑性。采用基因本体论、基因集富集分析和基因集变异分析对 CC 中的 MC 亚群进行差异表达分析。 CellChat 软件包用于预测 MC 亚群和 CC 细胞之间的细胞通讯。细胞功能实验验证了 TNFRSF12A 在 HeLa 和 Caski 细胞系中的功能。此外，还构建了风险评分模型来评估各种风险评分在临床特征、预后、免疫浸润、免疫检查点和功能丰富方面的差异。利用拷贝数变异推断计算拷贝数变异水平。获得的93,524个高质量细胞被分为十种细胞类型，包括T_NK细胞、内皮细胞、成纤维细胞、平滑肌细胞、上皮细胞、B细胞、浆细胞、MC 、中性粒细胞和骨髓细胞。此外，总共 1,392 个 MC 被细分为 7 个亚群：C0 CTSG MC、C1 CALR MC、C2 ALOX5 MC、C3 ANXA2 MC、C4 MGP MC、C5 IL32 MC 和 C6 ADGRL4 MC。值得注意的是，C2亚群与肿瘤相关MCs表现出密切的关联，Slingshot结果表明C2亚群处于分化的中晚期，可能代表CC良性向恶性转化的关键转变点。 CNVscore和批量分析结果进一步证实了C2亚群的转化状态。 CellChat 分析表明 TNFRSF12A 是参与 C2 ALOX5 MC 活动的关键受体。此外，体外实验表明下调TNFRSF12A基因可能部分抑制CC的发展。此外，基于C2亚群标志基因的预后模型和免疫浸润分析为患者预后和临床干预策略提供了有价值的指导。我们首先鉴定了CC内转化性肿瘤相关MCs亚群C2 ALOX5 MCs，该亚群处于CC的关键期。肿瘤分化阶段并影响 CC 的进展。体外实验证实敲除TNFRSF12A基因对CC的发展具有抑制作用。基于C2 ALOX5 MCs子集构建的预后模型表现出优异的预测价值。这些发现为 CC 的临床决策提供了新的视角。版权所有 © 2024 赵、洪、周、黄、林、张、梁和唐。

Cervical cancer (CC) is the fourth most common malignancy among women globally and serves as the main cause of cancer-related deaths among women in developing countries. The early symptoms of CC are often not apparent, with diagnoses typically made at advanced stages, which lead to poor clinical prognoses. In recent years, numerous studies have shown that there is a close relationship between mast cells (MCs) and tumor development. However, research on the role MCs played in CC is still very limited at that time. Thus, the study conducted a single-cell multi-omics analysis on human CC cells, aiming to explore the mechanisms by which MCs interact with the tumor microenvironment in CC. The goal was to provide a scientific basis for the prevention, diagnosis, and treatment of CC, with the hope of improving patients' prognoses and quality of life.The present study acquired single-cell RNA sequencing data from ten CC tumor samples in the ArrayExpress database. Slingshot and AUCcell were utilized to infer and assess the differentiation trajectory and cell plasticity of MCs subpopulations. Differential expression analysis of MCs subpopulations in CC was performed, employing Gene Ontology, gene set enrichment analysis, and gene set variation analysis. CellChat software package was applied to predict cell communication between MCs subpopulations and CC cells. Cellular functional experiments validated the functionality of TNFRSF12A in HeLa and Caski cell lines. Additionally, a risk scoring model was constructed to evaluate the differences in clinical features, prognosis, immune infiltration, immune checkpoint, and functional enrichment across various risk scores. Copy number variation levels were computed using inference of copy number variations.The obtained 93,524 high-quality cells were classified into ten cell types, including T_NK cells, endothelial cells, fibroblasts, smooth muscle cells, epithelial cells, B cells, plasma cells, MCs, neutrophils, and myeloid cells. Furthermore, a total of 1,392 MCs were subdivided into seven subpopulations: C0 CTSG+ MCs, C1 CALR+ MCs, C2 ALOX5+ MCs, C3 ANXA2+ MCs, C4 MGP+ MCs, C5 IL32+ MCs, and C6 ADGRL4+ MCs. Notably, the C2 subpopulation showed close associations with tumor-related MCs, with Slingshot results indicating that C2 subpopulation resided at the intermediate-to-late stage of differentiation, potentially representing a crucial transition point in the benign-to-malignant transformation of CC. CNVscore and bulk analysis results further confirmed the transforming state of the C2 subpopulation. CellChat analysis revealed TNFRSF12A as a key receptor involved in the actions of C2 ALOX5+ MCs. Moreover, in vitro experiments indicated that downregulating the TNFRSF12A gene may partially inhibit the development of CC. Additionally, a prognosis model and immune infiltration analysis based on the marker genes of the C2 subpopulation provided valuable guidance for patient prognosis and clinical intervention strategies.We first identified the transformative tumor-associated MCs subpopulation C2 ALOX5+ MCs within CC, which was at a critical stage of tumor differentiation and impacted the progression of CC. In vitro experiments confirmed the inhibitory effect of knocking down the TNFRSF12A gene on the development of CC. The prognostic model constructed based on the C2 ALOX5+MCs subset demonstrated excellent predictive value. These findings offer a fresh perspective for clinical decision-making in CC.Copyright © 2024 Zhao, Hong, Zhou, Huang, Lin, Zhang, Liang and Tang.