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.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54CA132381-07
Application #
8744917
Study Section
Special Emphasis Panel (ZCA1)
Project Start
Project End
Budget Start
2014-09-01
Budget End
2015-08-31
Support Year
7
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
City
Seattle
State
WA
Country
United States
Zip Code
98109
Tham, Heidi M; Hohl, Sarah; Copeland, Wade et al. (2016) Enhancing Biospecimen Knowledge Among Health Care Providers and Representatives From Community Organizations. Health Promot Pract :
Malen, Rachel; Knerr, Sarah; Delgado, Fernanda et al. (2016) Rural Mexican-Americans' perceptions of family health history, genetics, and disease risk: implications for disparities-focused research dissemination. J Community Genet 7:91-6
Salazar, Monica; Lerma-Ortiz, Alejandra; Hooks, Grace M et al. (2016) Progestin-mediated activation of MAPK and AKT in nuclear progesterone receptor negative breast epithelial cells: The role of membrane progesterone receptors. Gene 591:6-13
Sands, Bryan; Brent, Roger (2016) Overview of post Cohen-Boyer methods for single segment cloning and for multisegment DNA assembly. Curr Protoc Mol Biol 113:3.26.1-3.26.20
Briant, Katherine J; Wang, Lei; Holte, Sarah et al. (2015) Understanding the impact of colorectal cancer education: a randomized trial of health fairs. BMC Public Health 15:1196
Adams, Scott V; Barrick, Brian; Christopher, Emily P et al. (2015) Genetic variation in metallothionein and metal-regulatory transcription factor 1 in relation to urinary cadmium, copper, and zinc. Toxicol Appl Pharmacol 289:381-8
Yang, Fan; Kemp, Christopher J; Henikoff, Steven (2015) Anthracyclines induce double-strand DNA breaks at active gene promoters. Mutat Res 773:9-15
Knerr, Sarah; Ceballos, Rachel M (2015) Giving samples or ""getting checked"": measuring conflation of observational biospecimen research and clinical care in Latino communities. BMC Med Ethics 16:49
Briant, Katherine Josa; Espinoza, Noah; Galvan, Avigail et al. (2015) An innovative strategy to reach the underserved for colorectal cancer screening. J Cancer Educ 30:237-43
Thompson, Beti; Vilchis, Hugo; Moran, Crystal et al. (2014) Increasing cervical cancer screening in the United States-Mexico border region. J Rural Health 30:196-205

Showing the most recent 10 out of 44 publications