Dissecting the interplay between aging, genotype and the microenvironment in lung cancer

项目概要 癌症主要是老年人的疾病。虽然这部分是由于基因组的连续获取 除了变化之外，衰老还与一系列可能影响肿瘤发生和生长的变化有关。 这些“衰老标志”涉及多种影响致癌作用并导致系统性癌症的途径。 变化。然而，尽管衰老与癌症之间存在非常密切的联系，或者也许正因为如此，非常 关于癌细胞内在的、微环境的和全身年龄相关的变化如何影响我们知之甚少 癌症的发生和发展。基因工程小鼠模型独特地能够引入定义的 具有明确时间控制的正常成体细胞的遗传改变。人类肺癌模型已建立 使用基因工程小鼠模型，这些肿瘤重现了早期人类的许多特征 肺腺癌。为了提高体内癌症模型的范围和精度，我们集成了 传统的基因工程小鼠模型、基于 CRISPR/Cas9 的体细胞基因组工程，以及 使用统计方法的定量基因组学。肿瘤条形码与 CRISPR/Cas9 介导的基因相结合 灭活和高通量条形码测序 (Tuba-seq) 能够定量分析 与肿瘤起始和本地肿瘤生长的各个方面相关的大量基因。这些模型可以 从而区分衰老和突变事件的影响，同时提供一定程度的精确度，使我们能够 检测不同年龄背景下肿瘤抑制功能的差异。在目标 1 中，我们将量化交互 年龄和抑癌基因功能之间的关系。我们的体内实验将确定衰老是否会加剧 或降低肿瘤发生的绝对效率并揭示衰老对其重要性的影响 多种抑癌基因对肿瘤发生和生长的影响。在目标 2 中，我们将确定肺部肿瘤如何 微环境和肺癌细胞本身随着年龄的增长而变化。我们将阐明肿瘤的影响 跨年龄微环境的基因型，并确定年龄依赖性生长变化是否 伴随着癌细胞状态的巨大差异。在目标 3 中，我们将解开细胞自主性 肿瘤抑制功能驱动的年轻和老年小鼠肿瘤发展的差异 特别是通过局部组织和全身宿主环境的老化。这些实验将提供见解 研究年龄依赖性基因型特异性效应是否主要是癌细胞固有的或由肿瘤细胞的变化驱动的 微环境或整个有机体环境。通过改变癌细胞的年龄和基因型，以及 微环境和宿主年龄，我们将对这些因素的贡献获得前所未有的了解 肺癌发生的多个方面。最终，这些发现可能对癌症产生重要影响 预防、检测和治疗。

PROJECT SUMMARY Cancer is primarily a disease of the old. While this is due in part to the sequential acquisition of genomic alterations, aging is also associated with a constellation of changes that could impact tumor initiation and growth. These “hallmarks of aging” involve diverse pathways that impinge on carcinogenesis and lead to systemic changes. However, despite the very close association between aging and cancer, or perhaps because of it, very little is known about how cancer cell-intrinsic, microenvironmental, and systemic age-related changes impact cancer initiation and growth. Genetically engineered mouse models uniquely enable the introduction of defined genetic alterations into normal adult cells with defined temporal control. Human lung cancer has been modeled using genetically engineered mouse models, and these tumors recapitulate many features of early-stage human lung adenocarcinoma. To increase the scope and precision of in vivo cancer modeling, we integrated conventional genetically engineered mouse models, CRISPR/Cas9-based somatic genome engineering, and quantitative genomics with statistical approaches. Tumor barcoding coupled with CRISPR/Cas9-mediated gene inactivation and high-throughput barcode sequencing (Tuba-seq) enables quantitative analysis of the effects of large panels of genes on tumor initiation and various facets of autochthonous tumor growth. These models can thus distinguish the effects of aging from mutational events while affording a level of precision that allows us to detect differences in tumor suppressor function across age contexts. In Aim 1, we will quantify the interaction between age and tumor suppressor gene function. Our in vivo experiments will define whether aging increases or decreases the absolute efficiency of tumor initiation and uncover the impact of aging on the importance of diverse tumor suppressor genes on tumor initiation and growth. In Aim 2. we will determine how the lung tumor microenvironment and lung cancer cells themselves change with age. We will elucidate the impact of tumor genotype on the microenvironment across age and determine whether age-dependent changes in growth are accompanied by dramatic differences in cancer cell state. In Aim 3, we will disentangle cell-autonomous differences in tumors developing in young and aged mice from effects on tumor suppressor function driven specifically by aging of the local tissue and systemic host environments. These experiments will provide insight into whether age-dependent genotype-specific effects are largely cancer cell-intrinsic or driven by the shifts in the microenvironment or whole organism environment. By permuting cancer cell age and genotype, as well as microenvironment and host age, we will gain an unprecedented understanding of the contribution of these factors to multiple aspects of lung carcinogenesis. Ultimately, these findings could have important implication for cancer prevention, detection, and treatment.