Aleksandrovic Emilija, Fross Shaneann R, Golomb Samantha M, Liu Xiyu, Zhao Zhuo, Das Nikitha M, Reese Tanner C, Ma Wei, Lopez Jacqueline, Stack M Sharon, Zhao Min, Zhang Siyuan
Department of Pathology, UT Southwestern Medical Center, Dallas, TX, 75390, USA.
Harold C. Simmons Comprehensive Cancer Center, Dallas, TX, 75390, USA.
bioRxiv. 2025 Aug 17:2025.08.13.669778. doi: 10.1101/2025.08.13.669778.
Metastasis is an emergent continuum, driven by evolving reciprocal adaptations between continuously disseminating tumor cells (DTCs) and the specialized metastatic niches of distant organs. The interplay between intrinsic and niche-driven mechanisms that enables DTCs to survive and home to distant organs remains incompletely understood. Here, using MetTag, a single-cell barcoding and transcriptome profiling approach with time-stamped batch identifiers (BC.IDs), we mapped temporal, clonal dynamics of DTCs and the immune cell landscape across ovarian cancer metastatic niches. Deep sequencing of barcodes revealed preferred enrichment of early-disseminated clones across metastatic niches. Mechanistically, single-cell RNA sequencing (scRNA-seq) coupled with velocity analyses in ascites and metastasis-bearing omenta uncovered an emergent, distinct interferon-gamma (IFNγ) centric transcriptional trajectory, enriched among early seeding clones. Moreover, CRISPR/Cas9 screening of metastatic niche-specific signatures demonstrated that genes belonging to the ascites IFNγ signature, including , , are functionally important for peritoneal metastasis. Knockout of IFNγ receptor 1 () in tumor cells significantly reduced metastatic burden and extended survival, underscoring the importance of tumor cell intrinsic IFNγ signaling in ovarian cancer metastasis. Furthermore, we identified that the tumor intrinsic IFNγ response and ascites-derived tumor-associated macrophages (TAMs) protect cancer cells from anoikis-mediated death within the IFNγ-rich ascites environment. Our study resolves temporal dynamics of disseminating tumor cells and highlights an ascites-driven, IFNγ program as a necessary pro-metastatic adaptation in the ovarian metastasis cascade.
转移是一个渐进的连续过程，由不断播散的肿瘤细胞（DTCs）与远处器官的特殊转移微环境之间不断演变的相互适应性所驱动。使DTCs能够在远处器官存活并归巢的内在机制与微环境驱动机制之间的相互作用仍未完全了解。在这里，我们使用MetTag，一种带有时间戳批次标识符（BC.IDs）的单细胞条形码和转录组分析方法，绘制了DTCs的时间、克隆动态以及卵巢癌转移微环境中的免疫细胞图谱。条形码的深度测序揭示了早期播散克隆在转移微环境中的优先富集。从机制上讲，单细胞RNA测序（scRNA-seq）结合腹水和带转移灶的大网膜中的速度分析，发现了一个以干扰素-γ（IFNγ）为中心的新出现的独特转录轨迹，在早期播种克隆中富集。此外，对转移微环境特异性特征的CRISPR/Cas9筛选表明，属于腹水IFNγ特征的基因，包括 、 ，对腹膜转移具有重要功能。肿瘤细胞中干扰素-γ受体1（ ）的敲除显著降低了转移负担并延长了生存期，强调了肿瘤细胞内在IFNγ信号在卵巢癌转移中的重要性。此外，我们发现肿瘤内在的IFNγ反应和腹水来源的肿瘤相关巨噬细胞（TAMs）可保护癌细胞在富含IFNγ的腹水环境中免受失巢凋亡介导的死亡。我们的研究解析了播散肿瘤细胞的时间动态，并突出了腹水驱动的IFNγ程序作为卵巢转移级联中必要的促转移适应性。
