Defining age-associated alterations in oral squamous cell carcinoma

摘要 头颈癌鳞状细胞癌（HNSCC）是全球第七大常见癌症， 与存活率低有关。HNSCC的药物治疗在很大程度上仍然无效， 改善药物治疗需要鉴定不仅调节肿瘤而且调节肿瘤生长的新因子。 周围的口腔微环境。一个这样的新因素可能是组织的老化。而 年龄越大，HNSCC的5年生存率越差，几乎所有的临床前癌症模型研究 专注于年轻动物的结果。临床前青少年体内模型之间的这种不一致性， 老年目标患者人群可能是HNSCC治疗效果差的原因。这个目标 建议是定义与老年组织中HNSCC进展相关的分子事件， 发现可能提供新的治疗途径的靶向机制。为了测试年龄对HNSCC的影响， 我们优化了HNSCC的同基因原位舌异种移植模型。我们观察到肿瘤生长更快 在老年动物中，肿瘤表现出与年龄相关的、免疫逃避的、转录上不同的肿瘤细胞， 以及年龄相关的胶原分泌/修饰成纤维细胞群体。一个关键因素可能 与这些年龄相关的改变有关的是转录调节因子Yes相关蛋白（雅普）， 在HNSCC中具有促肿瘤发生作用的Hippo信号传导效应物，包括调节基因表达 追踪疾病进展和治疗抵抗。雅普介导转录变化， 促进攻击性细胞行为和免疫逃避，可能优先促进疾病的动力学 老年患者的严重程度。此外，老年动物的转录组显示了Runt的明显升高， 相关转录因子1（RUNX 1）在老年口腔肿瘤细胞。RUNX 1是一个雅普DNA结合伴侣， 被认为是HNSCC存活率更差的生物标志物。我们假设老化的口腔组织 微环境允许肿瘤发生，部分原因是细胞外基质动力学驱动细胞外基质的升高， 雅普和RUNX 1活性。在目标1中，我们将检查YAP缺失和RUNX 1缺失的肿瘤细胞在肿瘤细胞中的行为。 在我们的同基因原位杂交系统中，使用基因组、表观基因组和转录组学分析， 异种移植模型。从这一目标收集的数据将提供一个全面的比较的作用，雅普和 RUNX 1对HNSCC中观察到的介导年龄相关变化的影响。在目标2中，我们将定义年龄相关 老年口腔肿瘤小生境中淋巴细胞和骨髓免疫群体的变化，并测试 基质硬度对赋予这些年龄相关的变化，免疫和肿瘤细胞群体，在我们的研究中， 同系异种移植模型。从这一目标收集的数据将提供一个全面的评估年龄- 相关的免疫和机械组织靶点，用于HNSCC的潜在未来治疗。另外我们 拟议的计划提供了一个机会，在分子，细胞和遗传生物学，结合 与生物信息学和转化医学，磨练所需的成功作为一个医生，科学家的技能。

Abstract Head and neck cancer squamous cell carcinomas (HNSCC) are the seventh most common cancers globally and are associated with poor survival rates. Pharmacological treatments for HNSCC remain largely ineffective and improving drug therapies requires the identification of novel factors that modulate not just the tumor but also the surrounding permissive oral microenvironment. One such novel factor may be the aging of the tissue. While older age is associated with worse five-year survival for HNSCC, almost all preclinical cancer model studies focus on outcomes in young animals. This discordance between preclinical adolescent in vivo models and largely elderly target patient populations may underlie the poor efficacy of treatment for HNSCC. The goal of this proposal is to define molecular events associated with HNSCC progression in aged tissues with the hope to uncover targetable mechanisms that may offer new therapeutic avenues. To test the influence of age on HNSCC, we optimized syngeneic orthotopic tongue xenograft models of HNSCC. We observed more rapid tumor growth in old animals, with tumors exhibiting an age-associated, immune evasive, transcriptionally-distinct tumor cell states as well as an age-associated collagen-secreting/modifying fibroblast population. A critical factor that may be involved in these age-associated alterations is the transcriptional regulator Yes-associated protein (YAP), a Hippo signaling effector that has a pro-tumorigenic role in HNSCC, including modulating the expression of genes that track with disease progression and treatment resistance. YAP mediates transcriptional changes that promote aggressive cell behavior and immune evasion, dynamics that may preferentially promote disease severity in older patients. Furthermore, the transcriptome of old animals revealed distinct elevation of the Runt- related transcription factor 1 (RUNX1) in aged oral tumor cells. RUNX1 is a YAP DNA-binding partner and has been implicated as a biomarker of worse HNSCC survival. We hypothesize that the aged oral tissue microenvironment is permissive to tumorigenesis in part due to extracellular matrix dynamics that drive elevated YAP and RUNX1 activity. In Aim 1, we will examine YAP-depleted and RUNX1-depleted tumor cell behavior in old and young mice using genomic, epigenomic and transcriptomic analyses in our syngeneic orthotopic xenograft models. Data collected from this aim will offer a comprehensive comparison of the role of YAP and RUNX1 on the mediating age-associated alterations observed in HNSCC. In Aim 2, we will define age-associated changes to lymphocytic and myeloid immune populations in the aged oral tumor niche and test the impact of matrix stiffness on conferring these age-associated alterations to immune and tumor cell populations in our syngeneic xenograft models. Data collected from this aim will provide a comprehensive assessment of age- associated immune and mechanical tissue targets for potential future therapeutics in HNSCC. In addition, our proposed plan provides an opportunity for diverse training in molecular, cellular and genetic biology, combined with bioinformatics and translational medicine, honing the skills required for success as a physician-scientist.