Cancer-prone cell states of the fallopian tubal epithelium

项目摘要 卵巢/子宫外高级别浆液性癌（HGSC）是卵巢癌中最常见和最具侵袭性的类型。 癌它通常在早期没有症状，超过80%的患者在晚期被诊断出来，通常 无法治愈的癌症阶段，当肿瘤已经转移时。广泛的整合基因组分析 允许鉴定几种临床上不同的HGSC亚型。HGSC的很大一部分来自于 输卵管上皮（TE）位于输卵管（又名子宫管或输卵管）的远端区域。最近单曲 远端TE的细胞转录组分析推断HGSC异质性可能与不同的细胞 存在于TE细胞谱系中的状态。不幸的是，精确的基于细胞谱系的TE细胞类型的层次结构 尚未建立。此外，TE的易患癌症的细胞状态没有充分定义， 影响这种状态的因素尚不清楚。因此，尚不清楚HGSC的临床病程是否不均匀， 细胞治疗反应的发展可能反映了这一过程的不同启动和进展模式。 恶性肿瘤我们的初步研究表明，除了已知的分泌型（OVGP 1+）和纤毛型（FOXJ 1+）外， CD 24+）上皮细胞，存在几种上皮细胞群体，其特征在于CD 24+的优先表达。 干/祖细胞标志物，如SLC 1A 3、CD 49 f（ITGA 6）和KLF 6。一个Monocle细胞谱系轨迹 我们的单细胞转录组学数据的预测分析鉴定了一群SLC 1A 3+干/祖细胞 通过短暂的中间体，包括 KRT 5+细胞群。这一预测已经通过SLC 1A 3+细胞的谱系追踪和离体证实。 问题研究处于瞬时状态的细胞（CD 24 med CD 49 f+）在连续几轮的球体解离中形成球体- 再生并表达KRT 5。在正常的稳态条件下，KRT 5+细胞大部分处于休眠状态， 对分泌和纤毛细胞谱系的贡献最小。然而，KRT 5+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，基质电荷开始共- 在最早的形态学可检测到的改变之前与突变的上皮细胞一起进化。基于先前 研究和我们的初步结果，我们假设，癌症倾向TE细胞状态是由水平的 上皮损伤和基质环境改变。为了验证这一假设，我们提出（1）建立的作用， 在稳态和创伤后再生过程中的特定细胞状态，（2）确定上皮细胞的影响， 损害上皮状态的癌症易感性和（3）识别和表征上皮和基质细胞 TE恶性转化早期阶段的谱系动力学。

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.