单细胞转录组测序解析化疗后舌鳞癌免疫细胞动态图谱和肿瘤细胞耐药异质性  

Single-cell transcriptomic sequencing reveals the heterogeneity of immune and tumor cells in tongue squamous cell carcinoma after chemotherapy

作　　者：刘俊延 冯芝恩[2] 伍文杰 刘慧宇 代玉松 李博 李天成[1,6] LIU Junyan;FENG Zhien;WU Wenjie;LIU Huiyu;DAI Yusong;LI Bo;LI Tiancheng(Department of Otorhinolaryngology,Head and Neck Surgery,Peking University First Hospital,Beijing 100034,China;Department of Oral and Maxillofacial-Head and Neck Oncology,Beijing Stomatological Hospital,Capital Medical University,Beijing 100050,China;Department of Oral and Maxillofacial Surgery,Peking University School and Hospital of Stomatology,Beijing100081,China;National Center of Stomatology&National Clinical Research Center for Oral Diseases,Beijing 100081,China;Central Laboratory,Peking University School and Hospital of Stomatology,Beijing 100081,China;Department of Otorhinolaryngology,Head and Neck Surgery,Beijing Chaoyang Hospital,Capital Medical University,Beijing 100069,China)

机构地区：[1]北京大学第一医院耳鼻咽喉头颈外科,北京100034 [2]首都医科大学附属北京口腔医院口腔颌面头颈肿瘤科,北京100050 [3]北京大学口腔医院口腔颌面外科,北京100081 [4]国家口腔医学中心·国家口腔疾病临床医学研究中心,北京100081 [5]北京大学口腔医学院口腔医院中心实验室,北京100081 [6]首都医科大学附属朝阳医院耳鼻咽喉头颈外科,北京100069

出　　处：《现代肿瘤医学》2024年第13期2318-2327,共10页Journal of Modern Oncology

基　　金：国家自然科学基金面上项目(编号:82172686);教育部中国高校产学研创新基金(编号:2022IT055);北京市卫健委适宜技术和成果推广项目(编号:BHTPP2022006);首都卫生发展科研专项(编号:2022-2-4076);北京市自然科学基金(编号:7212113)。

摘　　要：目的:探究舌鳞癌免疫和肿瘤细胞亚群的转录组异质性和化疗相关的变化。方法:研究收集了首都医科大学附属北京口腔医院口腔颌面头颈肿瘤科确诊舌鳞状细胞癌(简称舌鳞癌)、接受相同化疗方案却出现化疗疗效差异的2位患者的化疗前、后的肿瘤和外周血单核细胞样本共8例,进行单细胞转录组测序,并对测序结果进行数据分析。使用多重免疫荧光技术观察T细胞化疗前后变化和疗效出现差异的患者之间肿瘤细胞marker基因表达情况对比。结果:根据经典marker基因的表达将获得的细胞分成T细胞和NK细胞、B细胞和浆细胞、上皮细胞等12个主要的细胞亚群。数据分析结果显示,化疗后舌鳞癌免疫和肿瘤亚群细胞图谱发生了显著变化。2位化疗效果不同的患者在特定亚型的T细胞、NK细胞和髓样细胞表现出转录组异质性和化疗相关变化,多重免疫荧光实验发现化疗后舌鳞癌肿瘤微环境中Tregs细胞减少,而CTL细胞增加。此外,该研究探索了化疗前后肿瘤细胞的异质性,并定义了三组具有不同生物学特征和恶性程度的肿瘤细胞亚群。多重免疫荧光显示疗效较差者的原发肿瘤组织比疗效好者具有更多高度恶性组肿瘤细胞。结论:研究结果解析了化疗后舌鳞癌免疫和肿瘤细胞的异质性,对影响舌鳞癌化疗效果的因素提出了新的见解,有助于识别潜在免疫治疗靶点。Objective:To explore the transcriptomic heterogeneity and chemotherapy-related changes in immune and tumor cell subpopulations of tongue squamous cell carcinoma(TSCC).Methods:The study collected before and after chemotherapy tumor and peripheral blood mononuclear cell samples from two patients diagnosed with TSCC at the Department of Oral and Maxillofacial-Head and Neck Oncology,Beijing Stomatological Hospital,Capital Medical University.These patients received the same chemotherapy regimen but exhibited differences in chemotherapy efficacy.A total of 8 samples were subjected to single-cell transcriptomic sequencing,followed by data analysis of the sequencing results.Multiplex immunofluorescence techniques were used to observe changes in T cells before and after chemotherapy and to compare the expression of tumor cell marker genes between patients with different therapeutic outcomes.Results:Based on the expression of classical marker genes,the obtained cells were divided into 12 main cell subgroups,including T cells and NK cells,B cells and plasma cells,epithelial cells,and so on.Data analysis revealed the heterogeneity of immune and tumor cells in TSCC after chemotherapy.Two patients with different chemotherapy outcomes show transcriptome heterogeneity and chemotherapy-related changes in specific subtypes of T cells,NK cells and myeloid cells.Multiplex immunofluorescence experiments found a decrease in Tregs and an increase in CTL cells in the tumor tissue after chemotherapy.In addition,this study explored the heterogeneity of tumor cells before and after chemotherapy and defined three groups of tumor cell subgroups with different biological characteristics and degrees of malignancy.Multiplex immunofluorescence showed that the primary tumor tissue of patients with poorer therapeutic effects had more highly malignant tumor cells compared to those with better therapeutic outcomes.Conclusion:The research results revealed the heterogeneity of immune and tumor cells in TSCC,offering new insights into the factors affec

关 键 词：单细胞转录组测序 肿瘤异质性 舌鳞状细胞癌 化疗 

分 类 号：R739.86[医药卫生—肿瘤]