(PQ1) Characterization of Premalignant Fields in a Murine Model of Head and Neck and Esophageal Cancers

(PQ1) 头颈癌和食管癌小鼠模型癌前区域的表征

基本信息

批准号：
9903826
负责人：
LORRAINE J GUDAS
金额：
$ 9.7万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-23 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9903826
关键词：
Address Affect Applications Grants Carcinogens Cells Chemopreventive Agent Clone Cells DNA sequencing Development Early treatment Enterobacteria phage P1 Cre recombinase Epigenetic Process Epithelial Epithelial Cells Epithelium Esophageal Esophageal Squamous Cell Carcinoma Esophagus Future Genetic Recombination Head Cancer Head and Neck Squamous Cell Carcinoma Human Incidence Individual LacZ Genes Larynx Malignant Neoplasms Malignant neoplasm of esophagus Measures Modeling Molecular Morphology Mucous Membrane Mus Mutation Neck Cancer Nitroquinolines Operative Surgical Procedures Oral cavity Oropharyngeal Oxides Parents Patients Pharmaceutical Preparations Physicians Population Premalignant Process Proliferating Property Proteins Radiation Relapse Stem cells Survival Rate Tamoxifen Testing Time Tobacco Transgenic Organisms cancer prevention carcinogenesis cell type chemotherapy design drinking water exome experimental study mouse model oral cavity epithelium progenitor stem targeted treatment transcriptome sequencing tumor whole genome

项目摘要

ABSTRACT FROM PARENT R01 (CA205258) Head and neck squamous cell carcinoma (HNSCC) occurs in the oral cavity, oropharynx, and larynx and is the 6th leading cancer in terms of incidence, affecting ~600,000 patients throughout the world. Esophageal squamous cell carcinoma (ESCC) is also quite common, with 17,000 new cases estimated in 2015 in the USA. Despite intensive treatment that generally combines surgery, radiation, and chemotherapy, HNSCCs and ESCCs relapse frequently and have poor long-term survival rates. Here we propose to use an approach using lineage-tracing to test the hypothesis that carcinogens cause early molecular changes in some progenitor/stem cells that reduce the diversity of the stem/progenitor cell population and subsequently result in both field cancerization and tumor formation in our murine HNSCC and ESCC carcinogenesis models. Thus, in this proposal we address one of the “Provocative Questions” of RFA-CA-15-008: For tumors that arise from a pre-malignant field, what properties of cells in this field can be used to design strategies to inhibit the development of future tumors? We will answer this question by performing two specific aims. Briefly, we will use doubly transgenic Keratin14/cre-ERTAM; Rosa26-lacZ mice to permanently mark individual normal stem/ progenitor cells and their progeny in the oral cavity and esophagus. Cre-dependent recombination will be activated only in cells that express K14 at the time of addition of tamoxifen (Tam), which is required for the cre-ERTAM protein to be active, giving us both temporal control and cell type-specific control over the cre recombinase activity and allowing us to follow the fates of these stem/ progenitor cells and their progeny over time. In Specific Aim (1), we will perform lineage-tracing, whole genome RNA-seq analyses, and whole exome DNA sequencing of these permanently marked lacZ+ “clones” of cells derived from individual, epithelial progenitor/stem cells during treatment with the carcinogen 4-nitroquinoline oxide (4-NQO), a tobacco surrogate, in the drinking water. This aim will delineate early molecular changes that occur in (a) the oral epithelium, and (b) the esophageal epithelium in a mouse model that very closely reflects human HNSCC and esophageal SCC development. We will also measure some key epigenetic changes. In Specific Aim (2) we will discern the key molecular changes that occur much later in the carcinogenesis process, when visible SCCs are present, by performing lineage tracing, whole genome RNA-seq analyses, and whole exome DNA sequencing of permanently marked “clones” of cells, derived from individual epithelial progenitor/ stem cells (both HNSCCs & ESCCs). In Aim (2) we will also compare tumors to clones of cells with normal morphology that are present near SCCs in “normal appearing” epithelial mucosa. Completion of these aims will define genetic and epigenetic changes that underlie ‘field cancerization,’ providing much new information about critical, early molecular changes in HNSCC and esophageal carcinogenesis that could be exploited to develop cancer chemopreventive drugs.

母公司 R01 的摘要 (CA205258) 头颈鳞状细胞癌 (HNSCC) 发生于口腔、口咽和喉部，就发病率而言，它是第六大癌症，影响全世界约 600,000 名患者。鳞状细胞癌 (ESCC) 也相当常见，2015 年美国估计有 17,000 例新病例。尽管强化治疗通常结合手术、放疗和化疗，但 HNSCC 和 ESCC 经常复发且长期生存率较差。在此，我们建议使用一种方法。谱系追踪以检验致癌物导致某些祖细胞/干细胞早期分子变化的假设减少干细胞/祖细胞群多样性并随后导致两个区域癌化的细胞以及我们的小鼠 HNSCC 和 ESCC 致癌模型中的肿瘤形成。因此，在本提案中，我们。解决 RFA-CA-15-008 的“挑衅性问题”之一：对于恶变前产生的肿瘤领域，该领域细胞的哪些特性可用于设计抑制发展的策略未来的肿瘤？我们将通过执行两个具体目标来回答这个问题。简单地说，我们将使用双重目标。转基因 Keratin14/cre-ERTAM；Rosa26-lacZ 小鼠永久标记个体正常干细胞/祖细胞口腔和食道中的细胞及其后代的Cre依赖性重组才会被激活。在添加他莫昔芬 (Tam) 时表达 K14 的细胞中，这是 cre-ERTAM 蛋白所必需的变得活跃，为我们提供了对 cre 重组酶活性的时间控制和细胞类型特异性控制，使我们能够随着时间的推移跟踪这些干/祖细胞及其后代的命运（1），我们将进行谱系追踪、全基因组 RNA-seq 分析和全外显子组 DNA 测序这些永久标记的 lacZ+ 细胞“克隆”源自个体上皮祖细胞/干细胞在使用致癌物 4-硝基喹啉氧化物 (4-NQO)（一种烟草替代品）进行治疗期间，该目标将描述（a）口腔上皮和（b）口腔中发生的早期分子变化。小鼠模型中的食管上皮非常接近人类 HNSCC 和食管 SCC 我们还将测量一些关键的表观遗传变化，在具体目标 (2) 中我们将辨别关键。当存在可见的鳞状细胞癌时，通过执行永久标记的谱系追踪、全基因组 RNA-seq 分析和全外显子组 DNA 测序细胞“克隆”，源自单个上皮祖细胞/干细胞（HNSCC 和 ESCC）。 (2) 我们还将比较肿瘤与存在于 SCC 附近的具有正常形态的细胞克隆。 “正常表现”的上皮粘膜的完成将定义遗传和表观遗传的变化。是“现场癌化”的基础，提供了许多关于关键的早期分子变化的新信息 HNSCC 和食管癌的发生可用于开发癌症化学预防药物。