Development Project 4

开发项目4

• 批准号: 8744899
• 负责人: BETI THOMPSON
• 金额: $ 17.11万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-25 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8744899
• 关键词: Address; Advanced Development; Algorithms; Animal Model; Area; Beds; Biochemical; Biological Assay; Cancerous; Cell Line; Cell Proliferation; Cell Therapy; Cell membrane; Cells; Clinical; Collaborations; Color; Complex; Confocal Microscopy; Data; Detection; Development; Effector Cell; Engineering; Event; Flow Cytometry; Fluorescence; Fred Hutchinson Cancer Research Center; Frequencies; Functional disorder; Funding; Future; Genotype; Green Fluorescent Proteins; International; Lead; Life; Location; Malignant Neoplasms; Mammalian Cell; Measurement; Measures; Mediating; Methods; Microscopic; Microscopy; New Mexico; Noise; Operating System; Phase; Phenotype; Population; Protein Engineering; Proteins; Recruitment Activity; Reporter; Research; Research Personnel; Saccharomyces cerevisiae; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Students; System; T-Lymphocyte; Technology; Testing; Time; Training; Tumor-Infiltrating Lymphocytes; Underrepresented Minority; Universities; Variant; Vision; Washington; Work; Yeast Model System; Yeasts; anticancer research; cancer diagnosis; cancer therapy; ethnic minority population; improved; instrument; instrumentation; interest; mutant; research study; response; time use; tumor; yeast genetics

We will make fully operational a continuing pilot collaboration between the labs of Dr. Jessica P. Houston (NMSU) and Dr. Roger Brent (FHCRC). Our proposed work builds on complementary strengths in the two labs: instrumentation engineering and signal analysis in the Houston lab, protein engineering and genetics in the Brent lab. In particular, the work builds on "frequency domain" methods to determine fluorescence lifetimes in microscopy, and on significantly more advanced developments in "frequency domain" methods for flow cytometry, for which Dr. Houston is emerging as a leader. Dysfunctions in cell signaling leading to inappropriate cell proliferation contribute to most cancers. Although much is known about cell signaling, key questions, including the causes and consequences of cell-to-cell variation in signaling and response, remain unanswered. Here, we will use fluorescent lifetime methods to extend the power of single cell assays to quantify key events in cell signaling in yeast (the "development platform") and in mammalian cells. These measurements depend on the use of Green Fluorescent Protein (GFP) derivatives fused to the proteins that make up the different cell signaling pathways to quantify events such as recruitment of a signaling complex to the cell membrane. During the next three years, we will develop our methods, to address these questions and to allow wider applications. In particular, we will continue to improve our instrumentation, use it to develop better GFP derivatives and signaling reporters, and use the improved methods to understand the causes of cell-to-cell variation in a yeast signaling system. We will also use these methods to develop high-signal reporters that allow quantification of cell signaling in clonal, mammalian cell lines using flow cytometry. Successful work will merit future funding. It should also increase the visibility of the Houston lab and help recruit new students and researchers from underrepresented ethnic minorities into an active area of international scientific research.

我们将使Jessica P. Houston博士（NMSU）和Roger Brent博士（FHCRC）实验室之间的持续试点合作全面运作。我们建议的工作建立在两个实验室的互补优势之上：休斯顿实验室的仪器工程和信号分析，布伦特实验室的蛋白质工程和遗传学。特别是，这项工作建立在“频域”方法上，以确定显微镜下的荧光寿命，以及流式细胞术的“频域”方法的显著更先进的发展，休斯顿博士正在成为这方面的领导者。