Development of Innovative Human Stromal Invasion Assay

创新型人类基质侵袭试验的开发

• 批准号: 6993325
• 负责人: B LYNN ALLEN-HOFFMANN
• 金额: $ 10万
• 依托单位: STRATATECH CORPORATION
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-29 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6993325
• 关键词: antineoplastics; bioassay; cell biology; cell line; clone cells; cytogenetics; drug discovery /isolation; drug screening /evaluation; fluorescence microscopy; gene therapy; head /neck neoplasm; high throughput technology; method development; molecular /cellular imaging; neoplasm /cancer invasiveness; nucleic acid sequence; pharmacokinetics; squamous cell carcinoma; stromal cells; tissue /cell culture; transfection /expression vector

DESCRIPTION (provided by applicant): The ultimate goal of this STTR proposal is to develop an innovative human tumor invasion assay for use as a screening tool in the identification of new anti-tumor therapeutics. Development of a model of early stage tumor invasion, the precursor condition to metastasis, coupled with a highly sensitive readout, is needed to identify new targets for drugs and/or agents that can be used to abrogate the invasion of primary H&N tumors and other types of carcinoma in humans. The NIKS(tm)-based human tumor model developed in the Allen-Hoffmann laboratory permits monitoring tumor cell growth within a normally developed, stratified squamous epithelium. Multiphoton imaging, an important tool for nondestructive investigation of living tissues, provides a highly sensitive readout for the detection of early changes in stroma underlying and/or surrounding tumors. We propose to expand upon our existing NIKS(tm)-NTM model by incorporating human tumor cells that possess a genetically engineered, invasive phenotype, and monitoring the behavior of these cells using multiphoton imaging. The following specific aims will be accomplished during Phase I: (1) Design and construct MT1-MMP expression vector, and demonstrate elevated MT1-MMP mRNA expression levels in transiently-transfected SCC13y cell monolayer cultures, (2) Demonstrate elevated protein expression levels and bioactivity in transiently-transfected SCC13y cell monolayer cultures, and (3) Develop stable, genetically-modified SCC13y cell clones, and evaluate bioactivity of these stable clones utilizing multiphoton microscopy. The unique combination of the novel stromal invasion model with a sophisticated imaging based readout will provide the pharmaceutical industry with a biologically relevant, human cell based, high throughput screening tool to evaluate novel cytostatic agents, and/or gene therapy strategies.

描述（由申请人提供）：该STTR提案的最终目标是开发一种创新的人类肿瘤入侵测定法，以用作鉴定新抗肿瘤疗法的筛查工具。需要开发早期肿瘤侵袭模型，需要转移的前体条件，再加上高度敏感的读数，以识别可用于废除原发性H＆N肿瘤的侵袭的药物和/或药物的新靶标，以及人类中其他类型的癌。基于NIKS（TM）的人类肿瘤模型在Allen-Hoffmann实验室中开发，允许监测正常发育的，分层的鳞状上皮的肿瘤细胞生长。多光子成像是对活组织的无损研究的重要工具，为检测抑制基质和/或周围肿瘤的早期变化提供了高度敏感的读数。我们建议通过结合具有遗传学工程，侵入性表型的人类肿瘤细胞来扩展现有的NIKS（TM）-NTM模型，并使用多光子成像监测这些细胞的行为。 The following specific aims will be accomplished during Phase I: (1) Design and construct MT1-MMP expression vector, and demonstrate elevated MT1-MMP mRNA expression levels in transiently-transfected SCC13y cell monolayer cultures, (2) Demonstrate elevated protein expression levels and bioactivity in transiently-transfected SCC13y cell monolayer cultures, and (3) Develop stable, genetically-modified SCC13Y细胞克隆，并利用多光显微镜检查这些稳定克隆的生物活性。新型基质入侵模型与基于成像成像的读数的独特组合将为制药行业提供具有生物学相关的，基于人类细胞的高吞吐量筛选工具，以评估新型的细胞抑制剂和/或基因治疗策略。