(PQ1) Progenitor cell malfunction, mutations and changes in microenvironment: A dynamic risk spectrum for cancer evolution

项目摘要/摘要 在这项提案中，我们将解决一个具有挑衅性的问题(PQ1)：“对于由 癌前野中细胞的什么特性可以用来设计策略来抑制 未来肿瘤的发展？ 我们将重点关注鳞状细胞肺癌(SCC)，这是一种常见的癌症死亡原因和 有相当多的证据支持起源于癌前领域的肿瘤。我们建议 要通过一项全面的战略来回答这个问题，该战略侧重于 肺鳞状细胞癌的癌前领域：上皮祖细胞功能，这是修复和 健康上皮的维持；癌前异型增生的基因组改变；以及组合物 和微环境的作用。我们假设“肺癌的进化是从一场 癌前异常增生症是由上皮祖细胞的内在变化以及 田间微环境发生的变化。因此，治疗可以针对抢救功能。 培养上皮祖细胞和/或恢复微环境。我们提出了三个目标来解决 这个假说。目标1.祖细胞功能障碍的机制，以确定新的靶点 化学预防：我们将确定上皮祖细胞是否在自我更新和 多潜能在发育不良谱系中各不相同。我们还将评估先祖对 化学预防性药物伊洛前列素在不同的不典型增生谱系中是不同的。RNA测序将探索 祖细胞功能障碍的决定因素和对伊洛前列素的反应，潜在地确定新的干预措施和 预测性生物标志物。目标2.检测新的基因组变化以确定新的靶点 化学预防：我们将评估不典型增生的遗传和表观遗传学特征是否在不同的 在异型增生谱系之上。从相同的发育不良谱系收集的刷子将用于识别 突变、表观遗传学改变和转录体改变；所有这些都与那些患有 SCC。将确定有针对性的突变和途径。目标3.组织结构和职能的变化 微环境以确定新的化学预防目标：我们将评估 微环境在上述不典型增生谱系中各不相同。将使用质量细胞术来表征 构成田间微环境的细胞。我们还将探索该领域中的细胞异质性 已知有持续性/进展史或单细胞退行性变病史的不典型增生的子集 流式分选上皮细胞和非上皮细胞的转录图谱。确定干预目标， 例如免疫检查点、炎症和血管生成将是我们的目标。我们预计我们的 综合治疗将导致新的策略，以防止癌前病变向侵袭性鳞癌的演变。

Project Summary/Abstract In this proposal we will address the provocative question (PQ1): “For tumors that arise from a premalignant field, what properties of cells in this field can be used to design strategies to inhibit the development of future tumors? We will focus on squamous cell lung cancer (SCC), a common cause of cancer mortality and a neoplasm for which there is considerable evidence supporting an origin from a premalignant field. We propose to answer this question through a comprehensive strategy that focuses on three critical elements of the premalignant field for SCC of the lung: epithelial progenitor cell function, which is required for repair and maintenance of a healthy epithelium; genomic alterations in premalignant dysplasias; and the composition and function of the microenvironment. We hypothesize that “Evolution of lung cancer from a field of premalignant dysplasia is driven both by intrinsic changes in epithelial progenitor cells as well as changes that occur in the field microenvironment. Thus, therapy could be targeted to rescue functions of epithelial progenitor cells and/or to restore the microenvironment.” We propose three aims to address this hypothesis. Aim 1. Mechanisms of progenitor malfunction to identify new targets for chemoprevention: We will determine whether epithelial progenitor malfunction in terms of self-renewal and multipotentiality varies across a dysplasia spectrum. We will also assess whether progenitor response to the chemopreventive agent iloprost differs across this dysplasia spectrum. RNA sequencing will explore determinants of progenitor dysfunction and response to iloprost, potentially identifying novel interventions and predictive biomarkers. Aim 2. Detecting novel genomic changes to identify new targets for chemoprevention: We will assess whether the genetic and epigenetic profile of dysplasia varies across the above dysplasia spectrum. Brushings collected from the same dysplasia spectrum will be used to identify mutations, epigenetic alterations and transcriptomal changes; all compared to the tumor in those subjects with SCC. Targetable mutations and pathways will be identified. Aim 3. Changes in composition and function of microenvironment to identify new targets for chemoprevention: We will assess whether the microenvironment varies across the above dysplasia spectrum. Mass cytometry will be used to characterize cells that constitute the field microenvironment. We will also explore cellular heterogeneity in the field in a subset of dysplasias with known history of persistence/progression or regression using single cell transcriptomic profiling of flow-sorted epithelial and non-epithelial cells. Identification of intervention targets, such as immune checkpoints, inflammation and angiogenesis will be our goal. We anticipate that our integrative approach will lead to new strategies to prevent the evolution of premalignancy to invasive SCC.