Recent developments in fluorescence microscopy geared for single-molecule detection, optical sectioning and three-dimensional data acquisition, used in combination with object-identification algorithms, now allow sufficient temporal and spatial resolution to visualize the sequential recruitment and intracellular traffic of proteins, lipids, and pathogens in living cells including viruses and bacteria. The Kirchhausen laboratory has a longstanding interest in understanding the mechanisms underlying entry of toxins, viruses and bacteria into cells, and how these processes relate to the organization and intracellular traffic of vesiculo-tubular membrane carriers. As part of this effort, we have devoted substantial efforts to developing an imaging suite containing state-of-the-art microscopes and supporting software suited for data collection and analysis with high spatial and temporal resolution. It is also a priority to maintain an imaging suite that is 100% accessible to all users, so that after initial training, any investigator can perform their work with total flexibility and independence. In this proposal we outline plans for a NERCE Imaging Core facility that will provide access to contemporary tools and expertise for live-cell and single molecule imaging aimed towards, but not limited to, quantitative descriptions of mechanisms of invasion for bacterial and viral pathogens into mammalian cells, of viral replication, and of molecular aspects related to toxin entry into cells.

Public Health Relevance

The Imaging Core will enable New England investigators to observe molecular events involving the entry, replication and pathology of infectious agents into host cells using real-time live cell imaging. The results of these studies will identify critical steps in the infectious process that may be targets for antimicrobial therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54AI057159-10
Application #
8441641
Study Section
Special Emphasis Panel (ZAI1-DDS-M)
Project Start
Project End
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
10
Fiscal Year
2013
Total Cost
$186,891
Indirect Cost
$58,024
Name
Harvard University
Department
Type
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
de Wispelaere, Melissanne; Lian, Wenlong; Potisopon, Supanee et al. (2018) Inhibition of Flaviviruses by Targeting a Conserved Pocket on the Viral Envelope Protein. Cell Chem Biol 25:1006-1016.e8
Huang, Nai-Jia; Pishesha, Novalia; Mukherjee, Jean et al. (2017) Genetically engineered red cells expressing single domain camelid antibodies confer long-term protection against botulinum neurotoxin. Nat Commun 8:423
Mertins, Philipp; Przybylski, Dariusz; Yosef, Nir et al. (2017) An Integrative Framework Reveals Signaling-to-Transcription Events in Toll-like Receptor Signaling. Cell Rep 19:2853-2866
Nair, Dhanalakshmi R; Chen, Ji; Monteiro, João M et al. (2017) A quinolinol-based small molecule with anti-MRSA activity that targets bacterial membrane and promotes fermentative metabolism. J Antibiot (Tokyo) 70:1009-1019
Choo, Min-Kyung; Sano, Yasuyo; Kim, Changhoon et al. (2017) TLR sensing of bacterial spore-associated RNA triggers host immune responses with detrimental effects. J Exp Med 214:1297-1311
de Wispelaere, Mélissanne; Carocci, Margot; Liang, Yanke et al. (2017) Discovery of host-targeted covalent inhibitors of dengue virus. Antiviral Res 139:171-179
Umetsu, Dale T (2017) Mechanisms by which obesity impacts upon asthma. Thorax 72:174-177
Zheng, Huiqing; Colvin, Christopher J; Johnson, Benjamin K et al. (2017) Inhibitors of Mycobacterium tuberculosis DosRST signaling and persistence. Nat Chem Biol 13:218-225
Coulson, Garry B; Johnson, Benjamin K; Zheng, Huiqing et al. (2017) Targeting Mycobacterium tuberculosis Sensitivity to Thiol Stress at Acidic pH Kills the Bacterium and Potentiates Antibiotics. Cell Chem Biol 24:993-1004.e4
Chiaraviglio, Lucius; Kang, Yoon-Suk; Kirby, James E (2016) High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity. J Vis Exp :

Showing the most recent 10 out of 417 publications