(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：
9303314
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/9303314
关键词：
Address Affect Alpha Cell Applications Grants Area Bioinformatics Carcinogens Cell Lineage Cells Chemopreventive Agent Clinical Treatment Clinical Trials Clone Cells DNA sequencing Development Dysplasia Early treatment Enterobacteria phage P1 Cre recombinase Epigenetic Process Epithelial Epithelium Esophageal Esophageal Squamous Cell Carcinoma Esophagus Faculty Functional disorder Funding Future Genetic Recombination Genomics Head Cancer Head and Neck Squamous Cell Carcinoma Head and Neck Surgery Head and neck structure 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 Neck Cancer Nitroquinolines Operative Surgical Procedures Oral Leukoplakia Oral cavity Oropharyngeal Otolaryngology Oxides Papilloma Pathology Patients Pharmaceutical Preparations Pharmacological Treatment Pharmacology Phenotype Physicians Population Positioning Attribute Premalignant Primary Neoplasm Process Proliferating Property Proteins Publishing Quality of life Radiation Recruitment Activity Recurrence Relapse Research Research Personnel Squamous cell carcinoma Statistical Data Interpretation Stem cells Stratum Basale Study models Survival Rate Tamoxifen Techniques Testing Time Tobacco Tongue Training Transgenic Organisms United States National Institutes of Health Universities cancer biomarkers cancer prevention carcinogenesis cell type chemotherapy cohesion conventional therapy design drinking water exome experience experimental study faculty research fluidity malignant mouth neoplasm medical schools meetings member molecular marker mouse model novel therapeutics oral cavity epithelium orofacial professor progenitor public health relevance stem success targeted treatment 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（由适用提供）：头部和颈部鳞状细胞癌（HNSCC）发生在口腔，口咽和喉部中，并且在发病率方面是第六个领先的癌症，影响了全球约60万名患者。食道鳞状细胞癌（ESCC）也很普遍，2015年在美国估计有17,000例新病例。尽管经过强化治疗通常结合了手术，放射线和化学疗法，但HNSCC和ESCC经常中继，长期存活率较差。在这里，我们建议使用一种使用谱系追踪的方法来检验以下假设：致癌物在某些祖细胞/干细胞中导致早期分子变化，从而减少了干/祖细胞细胞群的多样性，随后导致我们的Murine HNSCC和ESCC CCC CACCONEDONESENS模型导致现场取消和肿瘤形成。在此应用程序中，我们解决了RFA-CA-15-008的“挑衅性问题”之一：对于源自主体前场的肿瘤，该领域中细胞的特性可用于设计策略来抑制未来肿瘤的发展？我们将通过执行两个具体目标来回答这个问题。简而言之，我们将使用双重转基因角蛋白14/cre -etham； rosa26 − lacz小鼠在口腔和食道中永久标记单个正常的茎/祖细胞及其后代。 Cre依赖性重组仅在添加他莫昔芬（TAM）时表达K14的细胞中激活，这是Cre -ertam蛋白活跃所必需的，这使我们对Crece Recombinase活性进行了临时控制和细胞特异性控制，并允许我们遵循这些茎/幼虫细胞的损害时间。 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.这个目标将描绘（a）口服上皮的早期分子变化，以及（b）小鼠模型中的食管上皮，非常紧密地反映了人类HNSCC和食管SCC的发育。我们还将测量一些关键的表观遗传变化。 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).在目标（2）中，我们还将将肿瘤与具有正常形态的细胞克隆进行比较，这些细胞在“正常外观”上皮粘膜中存在于SCC附近。这些目的的完成将定义遗传和上皮变化，这些变化是“取消场取消”的基础，提供了有关HNSCC和食管癌变的关键，早期分子变化的许多新信息，可以探索这些信息以开发癌症化学预防药物。