Dysregulation of members of the epidermal growth factor (EGF) receptor (EGFR) family is associated with oncogenesis and tumor growth. Due to its relevance in cancer development, EGFR is an important target in cancer drug discovery, and several EGFR targeted therapies have already been developed. Their clinical success has, however, often been modest, reinforcing the need for a more complete understanding of the EGFR signaling pathway. This proposal focuses on the insufficiently understood role of large-scale EGFR associates ("clusters") in signaling initiation and transduction, in particular. While i has long been known that ligand induced dimerization plays a critical role in receptor signaling, there is growing evidence that this textbook model needs to be augmented to account for the heterogenous lateral distribution of the receptor in the plasma membrane. The local enrichment of the receptors in ''micro-domains" or ''nanoclusters''could strongly affect cooperative receptor interactions and shift the local EGFR association equilibria through a local concentration effect. The experimental investigation of the fundamental mechanisms underlying the large-scale receptor organization with conventional fluorescence microscopy remains challenging, due to the method's limitation with regard to throughput, spatial and temporal resolution, and maximum observation time. Plasmon Coupling Microscopy (PCM) is a novel non-fluorescence based approach that uses electromagnetic interactions between noble metal nanoparticles (NPs) to investigate receptor clustering on subdiffraction limit distances (but beyond the spatial barrier o Fluorescence Resonance Energy Transfer, FRET). NP's do not blink or bleach, are very bright, and can be imaged in a conventional widefield microscope. Consequently, PCM facilitates the monitoring of EGFR clustering without limitations in observation time in many individual cells simultaneously. This competitive renewal builds upon the plasmon coupling based tools developed in the previous funding cycle and outlines a vigorous research plan to elucidate the structural origin of dynamic EGFR clustering. PCM will then be applied to test the hypothesis that receptor clustering regulates the mode and strength of signaling and to elucidate the mechanisms underlying a spatial regulation of signaling intensity and outcome. The obtained insight will improve the understanding of spatial regulation mechanisms for a broad range of receptors. Noble metal NPs are not only superb optical labels for characterizing EGFR clustering in the plasma membrane, but they also represent potential therapeutic tools to restore and enhance negative EGFR signaling after covalent attachment to EGF. This hypothesis is experimentally tested in this proposal. If successful, this strategy would provide a new approach for overcoming apoptosis evasion in cancer.

Public Health Relevance

EGFR signaling is involved in regulating normal cell proliferation and tissue development. Abnormal EGFR signaling is a cause of uncontrolled cell growth in cancer. The proposed research will provide novel nanotechnologies that will make it possible to utilize the large-scale association of EGFR as a quantitative biomarker for the optical detection of dysregulated EGFR signaling on the single cell level. This project will improve patient outcomes and quality of life by advancing the molecular understanding of cancer and by enhancing early cancer detection and screening modalities.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
2R01CA138509-06
Application #
8578693
Study Section
Nanotechnology Study Section (NANO)
Program Officer
Knowlton, John R
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Boston University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02215
Wu, Linxi; Yu, Xinwei; Feizpour, Amin et al. (2014) Nanoconjugation: A Materials Approach to Enhance Epidermal Growth Factor Induced Apoptosis. Biomater Sci 2:156-166
Yu, Xinwei; Feizpour, Amin; Ramirez, Nora-Guadalupe P et al. (2014) Glycosphingolipid-functionalized nanoparticles recapitulate CD169-dependent HIV-1 uptake and trafficking in dendritic cells. Nat Commun 5:4136
Wu, Linxi; Reinhard, Björn M (2014) Probing subdiffraction limit separations with plasmon coupling microscopy: concepts and applications. Chem Soc Rev 43:3884-97
Chen, Tianhong; Pourmand, Mahshid; Feizpour, Amin et al. (2013) Tailoring Plasmon Coupling in Self-Assembled One-Dimensional Au Nanoparticle Chains through Simultaneous Control of Size and Gap Separation. J Phys Chem Lett 4:2147-2152
Puryear, Wendy Blay; Akiyama, Hisashi; Geer, Suzanne D et al. (2013) Interferon-inducible mechanism of dendritic cell-mediated HIV-1 dissemination is dependent on Siglec-1/CD169. PLoS Pathog 9:e1003291
Yu, Xinwei; Wang, Jing; Feizpour, Amin et al. (2013) Illuminating the lateral organization of cell-surface CD24 and CD44 through plasmon coupling between Au nanoparticle immunolabels. Anal Chem 85:1290-4
Chen, Tianhong; Reinhard, Björn M (2013) Characterizing the lateral friction of nanoparticles on on-chip integrated black lipid membranes. Small 9:876-84
Boriskina, Svetlana V; Reinhard, Bjorn M (2012) Molding the flow of light on the nanoscale: from vortex nanogears to phase-operated plasmonic machinery. Nanoscale 4:76-90
Ahn, Wonmi; Boriskina, Svetlana V; Hong, Yan et al. (2012) Electromagnetic field enhancement and spectrum shaping through plasmonically integrated optical vortices. Nano Lett 12:219-27
Rong, Guoxin; Reinhard, Björn M (2012) Monitoring the size and lateral dynamics of ErbB1 enriched membrane domains through live cell plasmon coupling microscopy. PLoS One 7:e34175

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