Abstract：
High-grade intraepithelial neoplasia (HIN) is the precursor of oesophageal squamous cell carcinoma. The molecular and functional properties of HIN are determined by intrinsic origin cells and the extrinsic microenvironment. Yet, these factors are poorly understood.
We performed single-cell RNA sequencing of cells from HINs and adjacent tissues from the human oesophagus. We analysed the heterogeneity of basal layer cells and confirmed it using immunostaining. Aneuploid cells in HIN were studied using primary cell culture combined with karyotype analysis. We reconstructed the lineage relationship between tumour and normal populations based on transcriptome similarity. Integration analysis was applied to our epithelial data and published invasive cancer data, and results were confirmed by immunostaining and 3D organoid functional experiments. We also analysed the tumour microenvironment of HIN.
The basal layer contained two cell populations: KRT15high STMN1low and KRT15high STMN1high cells, which were located mainly in the interpapillary and papillary zones, respectively. The KRT15high STMN1low population more closely resembled stem cells and transcriptome similarity revealed that HIN probably originated from these slow-cycling KRT15high STMN1low cells. 3D Organoid experiments and RNA-sequencing showed that basal-cell features and the differentiation ability of the normal epithelium were largely retained in HIN, but may change dramatically in tumour invasion stage. Moreover, the tumour microenvironment of HIN was characterised by both inflammation and immunosuppression.
Our study provides a comprehensive single-cell transcriptome landscape of human oesophageal HIN. Our findings on the origin cells and unique microenvironment of HIN will allow for the development of strategies to block tumour progression and even prevent cancer initiation.
We performed single-cell RNA sequencing of cells from HINs and adjacent tissues from the human oesophagus. We analysed the heterogeneity of basal layer cells and confirmed it using immunostaining. Aneuploid cells in HIN were studied using primary cell culture combined with karyotype analysis. We reconstructed the lineage relationship between tumour and normal populations based on transcriptome similarity. Integration analysis was applied to our epithelial data and published invasive cancer data, and results were confirmed by immunostaining and 3D organoid functional experiments. We also analysed the tumour microenvironment of HIN.
The basal layer contained two cell populations: KRT15high STMN1low and KRT15high STMN1high cells, which were located mainly in the interpapillary and papillary zones, respectively. The KRT15high STMN1low population more closely resembled stem cells and transcriptome similarity revealed that HIN probably originated from these slow-cycling KRT15high STMN1low cells. 3D Organoid experiments and RNA-sequencing showed that basal-cell features and the differentiation ability of the normal epithelium were largely retained in HIN, but may change dramatically in tumour invasion stage. Moreover, the tumour microenvironment of HIN was characterised by both inflammation and immunosuppression.
Our study provides a comprehensive single-cell transcriptome landscape of human oesophageal HIN. Our findings on the origin cells and unique microenvironment of HIN will allow for the development of strategies to block tumour progression and even prevent cancer initiation.
摘要：
高级别上皮内瘤变（HIN）是食管鳞状细胞癌的前体。HIN的分子和功能特性由内在起源细胞和外在微环境决定。然而，这些因素知之甚少。
我们对来自人食管的HIN和邻近组织的细胞进行了单细胞RNA测序。我们分析了基底层细胞的异质性，并使用免疫染色进行了确认。使用原代细胞培养结合核型分析研究了HIN中的非整倍体细胞。我们基于转录组相似性重建了肿瘤和正常人群之间的谱系关系。整合分析应用于我们的上皮数据和发表的浸润性癌症数据，结果通过免疫染色和3D类器官功能实验得到证实。我们还分析了HIN的肿瘤微环境。
基底层包含两种细胞群：KRT15highSTMN1low和KRT15highSTMN1high细胞，主要位于乳头间和乳头区，分别。KRT15highSTMN1low群体更接近干细胞，转录组相似性表明HIN可能起源于这些缓慢循环的KRT15highSTMN1low细胞。3D类器官实验和RNA测序表明，HIN基本上保留了正常上皮的基底细胞特征和分化能力，但在肿瘤侵袭阶段可能会发生巨大变化。此外，HIN的肿瘤微环境以炎症和免疫抑制为特征。
我们的研究提供了人类食道HIN的全面单细胞转录组景观。我们对HIN的起源细胞和独特微环境的发现将允许开发阻断肿瘤进展甚至预防癌症发生的策略。
我们对来自人食管的HIN和邻近组织的细胞进行了单细胞RNA测序。我们分析了基底层细胞的异质性，并使用免疫染色进行了确认。使用原代细胞培养结合核型分析研究了HIN中的非整倍体细胞。我们基于转录组相似性重建了肿瘤和正常人群之间的谱系关系。整合分析应用于我们的上皮数据和发表的浸润性癌症数据，结果通过免疫染色和3D类器官功能实验得到证实。我们还分析了HIN的肿瘤微环境。
基底层包含两种细胞群：KRT15highSTMN1low和KRT15highSTMN1high细胞，主要位于乳头间和乳头区，分别。KRT15highSTMN1low群体更接近干细胞，转录组相似性表明HIN可能起源于这些缓慢循环的KRT15highSTMN1low细胞。3D类器官实验和RNA测序表明，HIN基本上保留了正常上皮的基底细胞特征和分化能力，但在肿瘤侵袭阶段可能会发生巨大变化。此外，HIN的肿瘤微环境以炎症和免疫抑制为特征。
我们的研究提供了人类食道HIN的全面单细胞转录组景观。我们对HIN的起源细胞和独特微环境的发现将允许开发阻断肿瘤进展甚至预防癌症发生的策略。
