PO.TB10.01 · 肿瘤生物学
输卵管和卵巢的单细胞转录组分析以定义细胞类型及BRCA特异性标志物
Single-cell transcriptomic analysis of fallopian tube and ovary to define cell type and BRCA-specific markers
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
大多数高级别浆液性癌被认为起源于输卵管上皮，并通过在BRCA1/2突变携带者中更易发生转化的癌前状态进展。然而，转化过程中出现的早期上皮和基质程序仍未得到充分定义。为填补这一空白，我们分析了来自28例患者的49个FFPE scRNA-seq样本，包括正常卵巢和输卵管组织、BRCA1/2癌前组织，以及配对的原发卵巢肿瘤和转移病灶。FASTQ文件通过Cell Ranger流程比对到GRCh38，随后进行双细胞去除。对正常和癌前卵巢或输卵管样本，以及原发肿瘤和转移样本分别使用相互主成分分析进行整合。细胞类型注释以一个源自公开人输卵管scRNA-seq数据集的定制参考为指导，并使用经典标志基因进行验证。在各样本中，我们鉴定出主要的上皮、免疫和基质区室，包括不同的分泌型和纤毛型上皮群。早期分析显示，来自原发肿瘤和转移病灶的恶性上皮细胞绝大多数映射到分泌型上皮谱系，纤毛细胞贡献极小。正常和BRCA1/2癌前卵巢组织同样显示分泌型细胞的强烈富集，而输卵管组织则表现出更为均衡的分泌型-纤毛型分布。此外，与正常和癌前组织相比，原发和转移样本显示出显著增加的免疫浸润。这一全面的单细胞数据集为与BRCA1/2突变状态相关的早期上皮和微环境变化提供了新见解。正在进行的分析旨在确定特定的基质细胞状态（如成纤维细胞亚群）是否在癌前组织中出现并持续至恶性和转移病灶，提示存在早期促肿瘤基质表型。总体而言，本研究建立了一个用于发现卵巢癌起始的早期微环境驱动因素的框架。
查看英文原文 English abstract
Most high-grade serous carcinomas are thought to originate from the fallopian tube epithelium and progress through pre-malignant states that are more susceptible to transformation in BRCA1/2 mutation carriers. However, the early epithelial and stromal programs that arise during transformation remain poorly defined. To address this gap, we analyzed 49 FFPE scRNA-seq samples from 28 patients, including normal ovary and fallopian tube tissues, BRCA1/2 pre-malignant tissues, and matched primary ovarian tumors and metastatic lesions. FASTQ files were aligned to GRCh38 with the Cell Ranger pipeline, followed by doublet removal. Integration was performed using reciprocal principal component analysis separately for normal and pre-malignant ovary or fallopian tube samples, and primary tumor and metastatic samples. Cell type annotation was guided by a custom reference derived from publicly available human fallopian tube scRNA-seq datasets and validated using canonical marker genes. Across samples, we identified major epithelial, immune, and stromal compartments, including distinct secretory and ciliated epithelial populations. Early analyses revealed that malignant epithelial cells from both primary tumor and metastatic lesions overwhelmingly mapped to secretory epithelial lineages, with minimal contribution from ciliated cells. Normal and BRCA1/2 pre-malignant ovary tissues similarly showed a strong enrichment for secretory cells, whereas fallopian tube tissue displayed a more balanced secretory-ciliated distribution. In addition, primary and metastatic samples displayed markedly increased immune infiltration compared with normal and pre-malignant tissues. This comprehensive single-cell dataset provides new insight into early epithelial and microenvironmental changes associated with BRCA1/2 mutation status. Ongoing analyses aim to determine whether specific stromal cell states, such as fibroblast subpopulations, emerge in pre-malignant tissues and persist into malignant and metastatic lesions, suggesting the presence of early tumor-promoting stromal phenotypes. Overall, this study establishes a framework for discovering early microenvironmental drivers of ovarian carcinomas initiation.
利益披露 Disclosure
E. F. Medina, None..
J. I. Rodriguez, None..
A. Bujnak, None..
D. Lawson, None..
K. Kessenbrock, None.