Cancer-prone cell states of the fallopian tubal epithelium

项目摘要 卵巢/宫外高级别浆液性癌(HGSC)是最常见和最具侵袭性的卵巢癌 癌症。它通常在早期阶段没有症状，超过80%的患者被诊断为晚期，通常 无法治愈的癌症阶段，此时肿瘤已经转移。广泛的综合基因组分析 允许确定几个临床上不同的HGSC亚型。HGSC中很大一部分是由 输卵管上皮(TE)位于输卵管(又名输卵管)远端。最新单曲 细胞转录组分析推测HGSC的异质性可能与不同的细胞有关 在TE细胞谱系中存在状态。不幸的是，已识别的TE细胞类型的基于细胞谱系的精确层次结构 还没有建立起来。此外，TE易患癌症的细胞状态没有足够的定义，并且 影响这些州的因素仍不清楚。因此，目前尚不清楚HGSC的临床病程是否参差不齐。 细胞治疗反应的发展可能反映了不同的启动和进展模式 恶毒。我们的初步研究表明，除了已知的分泌型(OVGP1+)和纤毛型(FOXJ1+， CD24+)上皮细胞，有几种上皮细胞群的特点是优先表达 干细胞/祖细胞标记，如SLC1A3、CD49f(ITGA6)和KLF6。单细胞谱系轨迹 对我们的单细胞转录数据的预测分析确定了SLC1A3+干/祖细胞群体 通过瞬时中间产物产生分泌细胞和纤毛细胞，包括 Krt5+细胞群。这一预测已被SLC1A3+细胞的谱系追踪和体外实验所证实 学习。处于瞬时状态的细胞(CD24med CD49f+)在连续的几轮球体解离中形成球体- 再生并表达Krt5。在正常的动态平衡条件下，Krt5+细胞基本上处于休眠状态 对分泌和纤毛细胞谱系的贡献最小。然而，Krt5+细胞变得积极参与 机械损伤后再上皮化。我们的初步结果表明，间质电荷开始与 在最早的形态上可检测到的改变之前，与突变的上皮细胞一起进化。基于以前的 研究和我们的初步结果假设，易患癌症的TE细胞状态由以下因素决定 上皮损伤和间质环境改变。为了检验这一假设，我们建议(1)确定 在动态平衡和创伤后再生过程中特定的细胞状态，(2)确定上皮细胞的影响 上皮细胞状态对癌症易感性的损害以及(3)上皮细胞和基质细胞的鉴定和特征 TE恶变早期的谱系动力学研究。

Project Summary Ovarian/extra-uterine high-grade serous carcinoma (HGSC) is the most common and aggressive type of ovarian cancer. It often has no symptoms at early stages and over 80% of patients are diagnosed at advanced, usually incurable, cancer stages, when the tumors have already metastasized. Extensive integrated genomic analysis allowed identification of several clinically distinct subtypes of HGSC. A significant fraction of HGSC arises from the tubal epithelium (TE) located in the distal region of Fallopian tube (aka uterine tube or oviduct). Recent single cell transcriptome analysis of distal TE inferred that HGSC heterogeneity could be connected to diverse cell states present in TE cell lineages. Unfortunately, precise cell lineage-based hierarchy of identified TE cell types has not been yet established. Furthermore, cancer-prone cellular states of TE are insufficiently defined and factors influencing such states remain unclear. Thus, it remains unknown if uneven clinical course of HGSC and development of cellular therapeutic responses may reflect different modes of initiation and progression of this malignancy. Our preliminary studies show that, in addition to known secretory (OVGP1+) and ciliated (FOXJ1+, CD24+) epithelial cells, there are several epithelial cell populations characterized by preferential expression of stem/progenitor cell markers, such as SLC1A3, CD49f (ITGA6), and KLF6. A Monocle cell-lineage trajectory prediction analysis of our single-cell transcriptomic data identified a population of SLC1A3+ stem/progenitor cells that give rise to both secretory and ciliated cells by progressing through transient intermediates, including a KRT5+ cell population. This prediction has been confirmed by lineage tracing of SLC1A3+ cells and ex vivo studies. Cells in a transient state (CD24med CD49f+) form spheres in consecutive rounds of sphere dissociation- regeneration and express KRT5. Under normal homeostatic conditions, KRT5+ cells are largely dormant and minimally contribute to secretory and ciliated cell lineages. However, KRT5+ cells become actively involved in re-epithelialization after mechanical damage. Our preliminary results suggest that stromal charges begin to co- evolve with mutant epithelial cells before the earliest morphologically detectable alterations. Based on previous studies and our preliminary results we hypothesize that cancer-prone TE cell states are determined by levels of epithelial damage and stromal milieu changes. To test this hypothesis we propose (1) to establish the role of specific cell states during homeostatic and posttraumatic regeneration, (2) to determine the impact of epithelial damage on cancer susceptibility of epithelial states and (3) to identify and characterize epithelial and stromal cell lineage dynamics during early stages of TE malignant transformation.