金刚石瘤性颅咽管瘤（ACP）是一种临床上无有效治疗方法的侵袭性肿瘤。先前的研究提出了一种旁分泌肿瘤发生模型，其中致癌β-连环蛋白诱导垂体干细胞衰老，衰老细胞通过分泌促肿瘤因子导致旁分泌肿瘤的形成。然而，缺乏对 ACP 中衰老细胞的表征。在这里，我们利用单细胞 RNA 和 TCR 测序对 12 个 ACP 进行了分析，以阐明 ACP 中的细胞图谱，其中 3 个还进行了空间测序，以定位肿瘤细胞的不同亚群。我们总共获得了 70,682 个细胞的转录组图谱。通过驱动 CTNNB1 突变的细胞突变状态明确识别肿瘤细胞，将其分为 6 个子集。螺旋状簇 (WC) 细胞显示出与其他肿瘤细胞不同的分子特征，栅栏上皮 (PE) 细胞由增殖子集组成。除了典型的 PE 和 WC 之外，我们还发现了两个新的肿瘤细胞亚群。在一个亚群中，细胞表达高水平的细胞因子，例如 FDCSP 和 S100A8/A9，并富含衰老相关分泌表型 (SASP) 因子。苏木精和伊红染色显示这些 SASP 细胞缺乏有序结构，并且它们的细胞核被拉长。在另一个亚群中，细胞尺寸很小，并且紧密地堆积在一起，并且具有异常高的密度，表达高水平的线粒体基因（中位数 10.9%）。这些细胞是 RNA 速度和伪时间分析揭示的肿瘤发育轨迹的起源。单细胞 RNA 和 TCR 分析表明，一些 ACP 被克隆扩增的细胞毒性 T 细胞浸润。我们提出了一个假设，即 WC 和 PE 是通过过度激活的 WNT/β-catenin 信号传导的不同负调控机制形成的，这为 ACP 的肿瘤发生提供了新的认识。该研究为未来使用抗衰老化合物或其他治疗药物靶向 ACP 中衰老细胞的研究奠定了基础。© 2024。作者。

Adamantinomatous craniopharyngioma (ACP) is a clinically aggressive tumor without effective treatment method. Previous studies proposed a paracrine tumorigenesis model, in which oncogenic β-catenin induces senescence in pituitary stem cells and the senescent cells lead the formation of paracrine tumors through secretion of pro-tumorigenic factors. However, there lacks characterization on senescent cells in ACPs. Here, we profiled 12 ACPs with single-cell RNA and TCR-sequencing to elucidate the cellular atlas in ACPs and 3 of them were also subject to spatial sequencing to localize different subpopulations of the tumor cells. In total, we obtained the transcriptome profiles of 70,682 cells. Tumor cells, which were unambiguously identified through the cellular mutation status of the driver CTNNB1 mutations, were clustered into 6 subsets. The whorl-like cluster (WC) cells show distinct molecular features from the other tumor cells and the palisading epithelium (PE) cells consists of a proliferating subset. Other than typical PE and WC, we identified two novel subpopulations of the tumor cells. In one subpopulation, the cells express a high level of cytokines, e.g., FDCSP and S100A8/A9, and are enriched with the senescence-associated secretory phenotype (SASP) factors. Hematoxylin and eosin staining reveals that these SASP cells lack an ordered structures and their nuclei are elongated. In the other subpopulation, the cell sizes are small and they are tightly packed together with an unusual high density expressing a high level of mitochondrial genes (median 10.9%). These cells are the origin of the tumor developmental trajectories revealed by RNA velocity and pseudo-time analysis. Single-cell RNA and TCR analysis reveals that some ACPs are infiltrated with clonally expanded cytotoxic T cells. We propose a hypothesis that WC and PE are formed via different negative regulation mechanisms of the overactivated WNT/β-catenin signaling which provides a new understanding on the tumorigenesis of ACPs. The study lays a foundation for future studies on targeting senescent cells in ACPs with senolytic compounds or other therapeutic agents.© 2024. The Author(s).