PAI-1-mediated early-onset endometrial cancer

摘要 PAI-1介导的早发性子宫内膜癌 在过去的15年里，与肥胖相关的子宫内膜癌的发病率有所增加，这与此不谋而合 随着美国的全球肥胖症流行。这些患者比典型的患者年轻15-30岁。 因此，患者的临床负担与保留生育能力和疾病复发有关。至 了解肥胖对早发性子宫内膜癌的贡献，我们最近发现 肥胖患者子宫内膜微环境中脂肪基质细胞的分布初步 研究表明，ASCs直接影响细胞间相关基因座的表达变化 子宫内膜上皮细胞的通透性和极性(IPP)。单细胞转录组分图谱 进一步证实，纤溶酶原激活物抑制物-1(PAI-1)是一种由ASC分泌的丰富的脂肪因子，在 在解除对IPP职能的管制方面发挥关键作用。因此，我们假设异常的PAI-1信号干扰了 IPP转录，破坏细胞间通讯动态平衡，促进肿瘤EECS。在目标1中， ASC分泌的PAI-1在IPP转录重编程中的作用将在EEC中确定 曝光模型。当PAI-1与细胞表面的低密度脂蛋白受体相关蛋白1(LRP1)结合时， 内化信号产生E3泛素介导的Smad4降解，从而减弱转化生长因子β肿瘤- 抑制转录程序。单细胞蛋白质组图谱将确认IPP抑制 首发于年轻肥胖者原发肿瘤中Smad4低表达的细胞亚群 病人。在目标2中，将评估在EEC暴露中PAI-1对细胞转化的表型影响 带有IPP击倒或敲击的型号。染料转移分析和原子力显微镜将用于 探测EECS的细胞间通透性和极性特性，并确定这些特性是否发生了变化 生物物理特征代表EECS的一种肿瘤性表型。当在组织微阵列面板中验证时 230个肿瘤和30个未受累的正常样本(样本量合理)，候选基因表达降低 预计与高体重指数(BMI)患者的年轻年龄相关。在目标3中，PAI-1- DNA甲基转移酶的中介募集将在IPP易感基因座上进行检测。持续暴露 从EECS到PAI-1将促进甲基化在IPP启动子内的传播，留下永久的表观遗传 肿瘤后代的脚印。当在子宫内膜癌队列中得到证实时，DNA增加 这些候选基因的甲基化经常出现在年轻肥胖患者的原发肿瘤中。这个 拟议的研究不仅对PAI-1在早发性子宫内膜癌中的新作用提供了见解，而且还 识别表观遗传生物标记物，用于未来有发展风险的年轻患者的无细胞DNA监测 复发。

ABSTRACT PAI-1-mediated early-onset endometrial cancer Over the last 15 years, an increase in obesity-associated endometrial cancer has been observed, coinciding with the globesity epidemic in the US. These patients are 15-30 years younger than the typical patient demographics, and thus experience clinical burden linked to fertility preservation and disease recurrence. To understand the contribution of obesity to early-onset endometrial cancer, we recently found increased infiltration of adipose stromal cells (ASCs) in endometrial microenvironments of obese patients. Preliminary studies implicate that ASCs directly influence expression changes of loci associated with intercellular permeability and polarity (IPP) in endometrial epithelial cells (EECs). Single-cell transcriptomic profiling has further identified that plasminogen activator inhibitor-1 (PAI-1), an abundant ASC-secreted adipokine, plays a key role in deregulating IPP functions. Therefore, we hypothesize that aberrant PAI-1 signaling interferes with IPP transcription, disrupting intercellular communication homeostasis to promote neoplastic EECs. In Aim 1, the contribution of ASC-secreted PAI-1 to transcription reprogramming of IPP will be determined in EEC exposure models. When PAI-1 is tethered to LDL receptor-related protein 1 (LRP1) on the cell surface, the internalized signaling engenders E3 ubiquitin-mediated degradation of SMAD4 that attenuates TGFβ tumor- suppressive transcription program. Single-cell proteomic profiling will confirm whether IPP repression preferentially occurs in SMAD4-underexpressed cell subpopulations of primary tumors in young obese patients. In Aim 2, phenotypic influences of PAI-1 on cellular transformation will be assessed in EEC exposure models with IPP knockdowns or knockins. Dye-transfer assay and atomic force microscopy will be used to probe intercellular properties of permeability and polarity in EECs and to determine whether these altered biophysical features represent a neoplastic phenotype of EECs. When validated in a tissue microarray panel of 230 tumors and 30 uninvolved normal samples (sample size justified), decreased expression of candidate loci is expected to correlate with the young age of patients with high body mass indices (BMIs). In Aim 3, PAI-1- mediated recruitment of DNA methyltransferases will be examined in susceptible IPP loci. Persistent exposure of EECs to PAI-1 will facilitate methylation propagation within IPP promoters, leaving permanent epigenetic footprints in the neoplastic progeny. When confirmed in an endometrial cancer cohort, increased DNA methylation of these candidate loci is frequently present in primary tumors of young obese patients. The proposed study not only gives insights into a novel role of PAI-1 in early-onset endometrial cancer, but also identifies epigenetic biomarkers for cell-free DNA monitoring of young patients at risk of developing future recurrence.