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

剖析肺癌中衰老、基因型和微环境之间的相互作用

• 批准号: 10491833
• 负责人: Monte Meier Winslow
• 金额: $ 47.18万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10491833
• 关键词: Adult; Age; Aging; Architecture; Bar Codes; Biology; CRISPR/Cas technology; Cancer Cell Growth; Cancer Model; Carcinogens; Cell Line; Cells; Coupled; DNA Sequence Alteration; Data; Detection; Disease; Environment; Event; Gene Silencing; Genes; Genetically Engineered Mouse; Genome engineering; Genomics; Genotype; Goals; Growth; Human; Individual; KRAS2 gene; Lead; Lung Adenocarcinoma; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Methods; Modeling; Molecular; Mus; Mutation; Normal tissue morphology; Parabiosis; Pathway interactions; Positioning Attribute; Research Personnel; System; Tissues; Transplantation; Tumor Suppressor Genes; Tumor Suppressor Proteins; Whole Organism; age effect; age related; aged; base; cancer cell; cancer initiation; cancer prevention; carcinogenesis; cell age; experimental study; gene function; human cancer mouse model; human old age (65+); in vivo; insight; laboratory experience; lung cancer cell; lung carcinogenesis; novel; single cell analysis; theories; tumor; tumor growth; tumor initiation; tumor microenvironment; tumorigenesis

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.

项目摘要 癌症主要是旧的疾病。虽然这部分是由于基因组的顺序获得 改变，衰老也与可能影响肿瘤倡议和生长的变化星座有关。 这些“衰老的标志”涉及对癌变影响并导致全身性的各种途径 更改。但是，尽管衰老与癌症之间存在非常紧密的关联，或者也许是因为 关于癌细胞内膜，微环境和与年龄相关的变化如何影响癌症细胞中的变化鲜为人知 癌症倡议和增长。基因工程的鼠标模型独特地启用了定义的引入 具有定义的临时对照的正常成人细胞的遗传改变。人类肺癌已建模 使用一般设计的鼠标模型，这些肿瘤概括了早期人类的许多特征 肺腺癌。为了增加体内癌症建模的范围和精度，我们整合了 常规的基因工程鼠标模型，基于CRISPR/CAS9的体细胞基因组工程和 统计方法的定量基因组学。肿瘤条形码与CRISPR/CAS9介导的基因结合 灭活和高通量条形码测序（TUBA-SEQ）可以定量分析 大型基因肿瘤倡议和自肿瘤生长的各个方面。这些模型可以 因此，将衰老的影响与突变事件区分开 检测跨年龄范围的肿瘤抑制功能的差异。在AIM 1中，我们将量化相互作用 在年龄和肿瘤抑制基因功能之间。我们的体内实验将定义衰老是否增加 或降低肿瘤倡议的绝对效率，并发现衰老对 潜水员抑制肿瘤基因在肿瘤的启动和生长上。在AIM 2中。我们将确定肺肿瘤的方式 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 跨年龄的微环境的基因型，并确定生长的年龄依赖性变化是否为 伴随着癌细胞状态的巨大差异。在AIM 3中，我们将解散细胞自治 年轻小鼠和老年小鼠肿瘤对肿瘤抑制功能驱动的影响的差异 这些实验将提供洞察力 涉及年龄依赖性基因型特异性作用在很大程度上是癌细胞的内膜效应还是由转移驱动 微环境或整个生物环境。通过置换癌细胞的年龄和基因型以及 微环境和宿主时代，我们将对这些因素的贡献有前所未有的理解 到肺癌发生的多个方面。最终，这些发现可能对癌症具有重要意义 预防，检测和治疗。