(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：
9098367
负责人：
LORRAINE J GUDAS
金额：
$ 46.43万
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/9098367
关键词：
Address Affect Applications Grants Area Bioinformatics Carcinogens Cell Lineage Cells Chemopreventive Agent Clinical Treatment Clinical Trials Clone Cells DNA Sequence Development Dysplasia Early treatment Epigenetic Process Epithelial Epithelium Esophageal Esophageal Squamous Cell Carcinoma Esophagus Faculty Functional disorder Funding Future Genetic Recombination Genomics Head and Neck Cancer Head and Neck Squamous Cell Carcinoma Head and Neck Surgery Head and neck structure Health Health Care Research Health Policy Human Hyperplasia Incidence Individual Knowledge Laboratories LacZ Genes Larynx Lesion Leukoplakia Malignant Neoplasms Malignant neoplasm of esophagus Measures Modeling Molecular Molecular Abnormality Morphology Mucous Membrane Mus Mutation Nitroquinolines Normal Cell Oral Leukoplakia Oral cavity Oropharyngeal Otolaryngology Oxides Papilloma Pathology Patients Pharmaceutical Preparations Pharmacology Physicians Population Positioning Attribute Premalignant Primary Neoplasm Process Proliferating Property Proteins Publishing Quality of life Radiosurgery Recruitment Activity Recurrence Relapse Research Research Personnel Squamous cell carcinoma Statistical Data Interpretation Stem cells Stratum Basale Survival Rate Tamoxifen Techniques Testing Time Tobacco Tongue Training Transgenic Organisms United States National Institutes of Health Universities cancer prevention carcinogenesis cell type chemotherapy conventional therapy design drinking water exome experience faculty research fluidity malignant mouth neoplasm medical schools meetings member molecular marker mouse model novel therapeutics oral cavity epithelium orofacial professor progenitor recombinase research study stem success theories tool transcriptome sequencing tumor whole genome

项目摘要

DESCRIPTION (provided by applicant): 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 application 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.

描述（由申请人提供）：头颈鳞状细胞癌（HNSCC）发生在口腔、口咽和喉部，是发病率第六大的癌症，影响全球约 600,000 名 ESCC 患者。尽管强化治疗通常结合手术，但 2015 年美国估计有 17,000 例新病例。由于放疗和化疗，头颈部鳞状细胞癌和食管鳞癌经常复发，且长期存活率较低，因此我们建议使用谱系追踪方法来检验致癌物导致某些祖细胞/干细胞发生早期分子变化从而降低多样性的假设。干细胞/祖细胞群的变化以及随后在我们的小鼠 HNSCC 和 ESCC 致癌模型中导致现场癌化和肿瘤形成，因此，在本申请中，我们解决了“挑衅性问题”之一。 RFA-CA-15-008：对于恶变前区域产生的肿瘤，该区域细胞的哪些特性可用于设计抑制未来肿瘤发展的策略？我们将通过执行两个特定的操作来回答这个问题？简而言之，我们将使用双转基因 Keratin14/cre-ERTAM；Rosa26-lacZ 小鼠来永久标记口腔和食道中的个体正常干/祖细胞及其后代。仅在添加他莫昔芬 (Tam) 时表达 K14 的细胞中重组才会被激活，这是 cre-ERTAM 蛋白激活所必需的，从而为我们提供了对 cre 重组酶活性的时间控制和细胞类型特异性控制让我们能够追踪这些干细胞/祖细胞及其后代的命运在具体目标 (1) 中，我们将对这些来自个体上皮祖细胞/干细胞的永久标记的 lacZ+“克隆”在治疗过程中进行谱系追踪、全基因组 RNA 测序分析和全外显子组 DNA 测序。饮用水中的致癌物质 4-硝基喹啉氧化物 (4-NQO)（一种烟草替代品）。在与人类 HNSCC 和食管 SCC 发育密切相关的小鼠模型中，（a）口腔上皮和（b）食管上皮发生的早期分子变化我们还将测量一些关键的表观遗传变化。将通过进行谱系追踪、全基因组 RNA-seq 分析和永久的全外显子组 DNA 测序，识别致癌过程后期发生的关键分子变化（当存在可见的 SCC 时）标记的细胞“克隆”，源自单个上皮祖细胞/干细胞（HNSCC 和 ESCC）。在目标 (2) 中，我们还将肿瘤与存在于“正常外观”上皮中的 SCC 附近的具有正常形态的细胞克隆进行比较。完成这些目标将定义“现场癌化”背后的遗传和表观遗传变化，提供有关头颈部鳞状细胞癌和食管的关键早期分子变化的许多新信息。可用于开发癌症化学预防药物的致癌作用。