Immunocompromise of the T cell response may facilitate the aging process; thus, understanding the spatio-temporal dynamics of signaling in T cells at the molecular level is crucial. We hypothesize that cholesterol-rich membrane domains (or ?rafts?) facilitate functionally relevant compartmentalization for effective T cell signaling, and these critical molecular interactions between the T cell receptor and other signaling proteins and specific lipids are disrupted as aging proceeds. Cholesterol-rich rafts are hypothesized to be transient and microscopic and, until now, have not yet been directly observed in vivo without significant perturbation. This interdisciplinary NIH R21 proposal will overcome this problem by combining cutting-edge imaging tools with experimental approaches that will allow us to understand quantitatively how molecules interact in biomembranes. To test our hypothesis, we propose the following two Specific Aims:
Specific Aim 1 : Investigating the signaling dynamics of lipid- and protein-induced functional domains in young and aging T cell models with high spatial and temporal resolution using one- and two-photon micro-spectroscopy;
and Specific Aim 2 : Investigating the chemical nature of membrane domains that participate in immunoreceptor signaling using imaging mass spectrometry. For our initial studies on the latter Specific Aim, we will use the related model system of IgE receptor signaling in RBL mast cells, which serves as a robust platform for the development of imaging mass spectrometry on intact cells that we will subsequently use on the T cell models. The projected results will lead to new insights into how T cell receptor signaling dynamics and related plasma membrane nanostructure change as T cells age.? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AG030949-01
Application #
7334097
Study Section
Special Emphasis Panel (ZAG1-ZIJ-2 (M1))
Program Officer
Fuldner, Rebecca A
Project Start
2007-09-01
Project End
2009-06-30
Budget Start
2007-09-01
Budget End
2008-06-30
Support Year
1
Fiscal Year
2007
Total Cost
$102,027
Indirect Cost
Name
Pennsylvania State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
003403953
City
University Park
State
PA
Country
United States
Zip Code
16802
Currie, Megan; Leopold, Hannah; Schwarz, Jacob et al. (2017) Fluorescence Dynamics of a FRET Probe Designed for Crowding Studies. J Phys Chem B 121:5688-5698
Heikal, Ahmed A (2014) Time-resolved fluorescence anisotropy and fluctuation correlation analysis of major histocompatibility complex class I proteins in fibroblast cells. Methods 66:283-91
Solntsev, Pavlo V; Spurgin, Katelynn L; Sabin, Jared R et al. (2012) Photoinduced charge transfer in short-distance ferrocenylsubphthalocyanine dyads. Inorg Chem 51:6537-47
Heikal, Ahmed A (2010) Intracellular coenzymes as natural biomarkers for metabolic activities and mitochondrial anomalies. Biomark Med 4:241-63
Vats, Kanika; Knutson, Kristofer; Hinderliter, Anne et al. (2010) Peripheral protein organization and its influence on lipid diffusion in biomimetic membranes. ACS Chem Biol 5:393-403
Ariola, Florly S; Li, Zaiguo; Cornejo, Christine et al. (2009) Membrane fluidity and lipid order in ternary giant unilamellar vesicles using a new bodipy-cholesterol derivative. Biophys J 96:2696-708
Piehowski, Paul D; Davey, Angel M; Kurczy, Michael E et al. (2009) Time-of-flight secondary ion mass spectrometry imaging of subcellular lipid heterogeneity: Poisson counting and spatial resolution. Anal Chem 81:5593-602
Yu, Qianru; Heikal, Ahmed A (2009) Two-photon autofluorescence dynamics imaging reveals sensitivity of intracellular NADH concentration and conformation to cell physiology at the single-cell level. J Photochem Photobiol B 95:46-57
Kyoung, Minjoung; Sheets, Erin D (2008) Vesicle diffusion close to a membrane: intermembrane interactions measured with fluorescence correlation spectroscopy. Biophys J 95:5789-97
Vats, Kanika; Kyoung, Minjoung; Sheets, Erin D (2008) Characterizing the chemical complexity of patterned biomimetic membranes. Biochim Biophys Acta 1778:2461-8

Showing the most recent 10 out of 13 publications