宫颈癌(CC)是全球女性中第四大常见恶性肿瘤,是发展中国家女性癌症相关死亡的主要原因。CC的早期症状往往不明显,诊断通常是在晚期阶段进行的,导致不良的临床预后。近年来,大量研究表明,肥大细胞(MCs)与肿瘤的发生发展有着密切的关系。然而,目前对MC在CC中所起作用的研究还非常有限。因此,这项研究对人类CC细胞进行了单细胞多组学分析,目的探讨MC与CC中肿瘤微环境相互作用的机制。目的是为预防提供科学依据,诊断,和CC的治疗,希望改善患者预后和生活质量。
本研究从ArrayExpress数据库中的十个CC肿瘤样品获得单细胞RNA测序数据。弹弓和AUCcell用于推断和评估MC亚群的分化轨迹和细胞可塑性。CC中MCs亚群的差异表达分析,采用基因本体论,基因集富集分析,和基因集变异分析。应用CellChat软件包预测MC亚群与CC细胞之间的细胞通讯。细胞功能实验验证了HeLa和Caski细胞系中TNFRSF12A的功能。此外,建立了一个风险评分模型来评估临床特征的差异,预后,免疫浸润,免疫检查点,以及各种风险评分的功能丰富。使用拷贝数变异的推断来计算拷贝数变异水平。
■将获得的93,524个高质量细胞分为十种细胞类型,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1392个MC被细分为七个亚群:C0CTSG+MC,C1CALR+MC,C2ALOX5+MC,C3ANXA2+MC,C4MGP+MC,C5IL32+MC,和C6ADGRL4+MC。值得注意的是,C2亚群与肿瘤相关的MC密切相关,弹弓结果表明C2亚群位于分化的中晚期,可能代表CC良性至恶性转化的关键过渡点。CNVscore和批量分析结果进一步证实了C2亚群的转化状态。CellChat分析显示TNFRSF12A是参与C2ALOX5+MC作用的关键受体。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亚群标记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.
■我们首先确定了CC内的转化性肿瘤相关MC亚群C2ALOX5+MC,处于肿瘤分化的关键阶段,并影响CC的进展。体外实验证实了敲低TNFRSF12A基因对CC发展的抑制作用。基于C2ALOX5+MC亚群构建的预后模型显示出优异的预测价值。这些发现为CC的临床决策提供了新的视角。
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.