Illuminating the Role of Oral Stem Cells in the Development of Oral Squamous Cell Carcinomas

阐明口腔干细胞在口腔鳞状细胞癌发展中的作用

基本信息

批准号：
9346053
负责人：
Antonio Luigi Amelio
金额：
$ 22.8万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
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：
2016-09-05 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9346053
关键词：
Address Adopted Affect Animal Cancer Model Biological Assay Biology Bioluminescence Cancer Biology Cancer Model Candidate Disease Gene Cell division Cells Chemicals Complex Critical Pathways Development Diagnostic Differentiated Gene Disease Energy Transfer Epigenetic Process Epithelial Epithelium Equilibrium Equipment and supply inventories Event Evolution Exhibits FAT gene Frequencies Gene Mutation Genes Genetic Genetic Techniques Genomics Goals Growth HPV-High Risk Head and Neck Squamous Cell Carcinoma Human Human Papillomavirus Human papillomavirus 16 Image Imaging Techniques Investments Kinetics Knowledge Lead Malignant Neoplasms Mediating Methods Modeling Mus Mutate Mutation NOTCH1 gene Oncoproteins Oral Papillomavirus Transforming Protein E7 Pathway interactions Patient-Focused Outcomes Patients Pharmacotherapy Play Principal Investigator Protocols documentation RNA Interference Reporter Research Resistance Resources Role Signal Pathway Skin Solid Neoplasm Stem cells Surface Survival Rate System Techniques Testing The Cancer Genome Atlas Therapeutic Time Transgenic Mice Translating Tumor Initiators Tumor stage addiction base cancer cell cancer recurrence chemical carcinogen combinatorial dimethylbenzanthracene effective therapy epigenetic regulation genetic manipulation genome sequencing genome-wide improved in utero in vivo innovation insight malignant mouth neoplasm mortality mouse model mouth squamous cell carcinoma neoplastic cell new therapeutic target next generation sequencing novel novel therapeutics optical imaging oral behavior oral carcinogenesis pluripotency prognostic tool self-renewal small hairpin RNA stem stemness targeted treatment therapeutic target tool transcription factor treatment strategy tumor tumor growth tumor initiation tumor progression tumorigenesis

项目摘要

SUMMARY Head and neck squamous cell carcinomas (HNSCCs) are the sixth most common cancer worldwide. Despite decades of research, afflicted patients continue to suffer a staggering ~50% five-year mortality rate. Two enormous obstacles impede the development of more effective therapies for HNSCC: 1) poor understanding of the pathways that lead to malignancy and 2) inability to detect tumors early enough. While much has been learned about the mutational landscape of HNSCCs through next generation sequencing, a tremendous challenge remains in translating this genomic information into functional relevance. Reliance on animal models of cancer is critical for separating driver from passenger mutations, to determine pathways that synergize to promote aggressive tumors, and to develop diagnostic/prognostic tools and therapeutic targets. Unfortunately, the ability to develop and rapidly test these models is hampered by the large investment of time and resources required to generate transgenic mice. This proposal addresses these significant knowledge gaps through the synergistic implementation of cutting-edge genetic and imaging techniques developed by the two principal investigators. Our long-term goal is to establish high-throughput mouse models of oral cancer that will elucidate cryptic pathways that promote tumor growth, providing insights into new avenues for therapy. Here, we adopt our versatile in utero lentiviral transduction technique and novel LumiFluor bioluminescence resonance energy transfer (BRET) reporter to a validated model of oral cancel, to manipulate underexplored genetic pathways revealed by genome-wide HNSCC sequencing studies, and visualize how they affect tumor growth kinetics. Our objective is to understand how self-renewal and differentiation pathways critical to normal development are co-opted by cancer cells. Our rationale is that two-thirds of HNSCC patient tumors show mutations in differentiation genes, which is correlated with poor patient survival. Based on our compelling preliminary studies, we will test the central hypothesis that imbalance between symmetric and asymmetric cell divisions in tumors—mediated by cytoskeletal control of the spindle orientation machinery and epigenetic regulation of “stem-ness”—play important roles in HNSCC tumorigenesis. Our Specific Aims are to: 1) Determine the function of spindle orientation genes in oral carcinogenesis, as this pathway directly regulates self-renewal/differentiation decisions; and 2) Establish a high-throughput in vivo functional assay for modulators of tumorigenesis. Both aims use our improved Cre-inducible mouse model of oral cancer based on a validated, established tumorigenesis protocol driven by high-risk HPV16 E6/E7 and the chemical carcinogen 4-NQO. The proposed research plan is both technically and conceptually innovative. The idea that spindle orientation is an important regulator of solid tumor growth kinetics is untested, and we possess a unique toolkit—in utero lentiviral RNAi for rapid genetic manipulations, LumiFluor reporter for advanced imaging, and an inducible model of HPV+ HNSCCs—that will allow us to make important insights into HNSCC biology.

概括头部和颈部鳞状细胞癌（HNSCCS）是全球第六个最常见的癌症。尽管数十年的研究，受苦的患者继续遭受惊人的五年死亡率。二巨大的障碍阻碍了针对HNSCC的更有效疗法的发展：1）对导致恶性肿瘤和2）无法尽早检测肿瘤的途径。虽然很多通过下一代测序了解HNSCC的突变景观，这是一个巨大的挑战仍然在将这些基因组信息转化为功能相关性。依赖动物模型癌症对于将驾驶员与乘客突变分开至关重要，以确定与促进侵袭性肿瘤，并开发诊断/预后工具和治疗靶标。很遗憾，大量的时间和资源投资阻碍了开发和快速测试这些模型的能力需要产生转基因小鼠。该建议通过两位主要的尖端遗传和成像技术的协同实施调查人员。我们的长期目标是建立高通量小鼠的口腔癌模型阐明促进肿瘤生长的加密途径，为新的治疗途径提供见解。这里，我们在子宫慢病毒转移技术和新型Lumifluor生物发光中采用多功能共振能量转移（BRET）记者到经过验证的口服取消模型，以操纵未置换全基因组HNSCC测序研究揭示了遗传途径，并可视化它们如何影响肿瘤生长动力学。我们的目标是了解自我更新和分化途径对正常发育由癌细胞选择。我们的理由是，HNSCC患者肿瘤的三分之二分化基因的突变与患者生存不良相关。基于我们引人注目的初步研究，我们将测试中心假设，即对称细胞和不对称细胞之间的失衡肿瘤中的划分 - 由纺锤体方向机制和表观遗传学的细胞骨架控制介导调节“干性” - 在HNSCC肿瘤发生中播放重要作用。我们的具体目的是：1）确定纺锤体取向基因在口服癌变中的功能，因为该途径直接调节自我更新/差异决定； 2）建立一个高通量的体内功能测定法肿瘤发生调节剂。两种目的都使用我们改进的Cre-Cre诱导小鼠口腔癌模型基于由高风险HPV16 E6/E7和化学致癌驱动的经过验证的，已建立的肿瘤发生协议 4-nqo。拟议的研究计划在技术上和概念上都是创新的。主轴的想法取向是未经测试的实体瘤生长动力学的重要调节剂，我们拥有独特的工具包 - 在子宫慢病毒RNAi中，用于快速遗传操纵，用于高级成像的Lumifluor报告基因和 HPV+ HNSCC的诱导模型 - 它将使我们能够对HNSCC生物学有重要见解。