Table_1_Revealing molecular and cellular heterogeneity in hypopharyngeal carcinogenesis through single-cell RNA and TCR/BCR sequencing.doc

IntroductionHypopharyngeal squamous cell carcinoma (HSCC) is one of the malignant tumors with the worst prognosis in head and neck cancers. The transformation from normal tissue through low-grade and high-grade intraepithelial neoplasia to cancerous tissue in HSCC is typically viewed as a progressive pathological sequence typical of tumorigenesis. Nonetheless, the alterations in diverse cell clusters within the tissue microenvironment (TME) throughout tumorigenesis and their impact on the development of HSCC are yet to be fully understood.
MethodsWe employed single-cell RNA sequencing and TCR/BCR sequencing to sequence 60,854 cells from nine tissue samples representing different stages during the progression of HSCC. This allowed us to construct dynamic transcriptomic maps of cells in diverse TME across various disease stages, and experimentally validated the key molecules within it.
ResultsWe delineated the heterogeneity among tumor cells, immune cells (including T cells, B cells, and myeloid cells), and stromal cells (such as fibroblasts and endothelial cells) during the tumorigenesis of HSCC. We uncovered the alterations in function and state of distinct cell clusters at different stages of tumor development and identified specific clusters closely associated with the tumorigenesis of HSCC. Consequently, we discovered molecules like MAGEA3 and MMP3, pivotal for the diagnosis and treatment of HSCC.
DiscussionOur research sheds light on the dynamic alterations within the TME during the tumorigenesis of HSCC, which will help to understand its mechanism of canceration, identify early diagnostic markers, and discover new therapeutic targets.

引言：下咽鳞状细胞癌（Hypopharyngeal squamous cell carcinoma, HSCC）是头颈部恶性肿瘤中预后最差的恶性肿瘤之一。HSCC中从正常组织经低级别、高级别上皮内瘤变进展为癌组织的过程，通常被认为是肿瘤发生的典型渐进性病理序列。然而，在整个肿瘤发生过程中，组织微环境（tissue microenvironment, TME）内不同细胞簇的变化及其对HSCC发生发展的影响，仍未被完全阐明。
方法：本研究采用单细胞RNA测序及TCR/BCR测序技术，对覆盖HSCC进展不同阶段的9份组织样本中的60854个细胞进行测序。借此构建了不同疾病阶段下不同TME中细胞的动态转录组图谱，并对其中的关键分子进行了实验验证。
结果：本研究阐明了HSCC肿瘤发生过程中肿瘤细胞、免疫细胞（包括T细胞、B细胞及髓系细胞）与基质细胞（如成纤维细胞、内皮细胞）之间的异质性；揭示了肿瘤发育不同阶段中不同细胞簇的功能与状态变化，并筛选出与HSCC肿瘤发生密切相关的特异性细胞簇，最终鉴定出MAGEA3、MMP3等对HSCC诊疗具有关键价值的分子。
讨论：本研究阐明了HSCC肿瘤发生过程中TME内的动态变化，有助于解析其癌变机制、筛选早期诊断标志物并发掘全新的治疗靶点。