Most bacteria in natural and clinical settings grow as surface-attached biofilms, which are bacterial communities that have self-assembled into an encased matrix of extracellular polymeric substances (EPS). To form these bacterial biofilm communities and infect host cells, an intercellular signaling process described as quorum sensing (QS) is very important. For the opportunistic pathogens Pseudomonas aeruginosa and Staphylococcus aureus, QS regulates the expression of many genes important to biofilm initiation, EPS production, and virulence. While much has been learned about select factors that regulate biofilm formation in vitro and in animal models, the specific mechanisms by which multispecies groups of cells form biofilms are not understood. Furthermore, the therapeutic capabilities needed to treat the increasingly virulent infections of the coming decades will require deeper insight into multi-species interactions and their regulation of multiple gene expression profiles as represented over extended spatial and temporal scales during host cell infection. The correlated mass spectrometric and Raman chemical imaging approaches that have been developed by our combined research group circumvent the limitations imposed by previous technology which allowed examination of either a small number of cells or entire cell populations that had been removed from the conditions of interest. In contrast our correlated chemical imaging methods allow the determination and spatial mapping of individual bacterial species and their microbial products within a mixed bacterial community growing in situ on surfaces. One of our long-term goals is the design of detection and diagnostic strategies informed by an understanding of bacterial interactions and signature biomolecule production. We will continue our work toward this goal by conducting mono-culture and co-culture experiments using Pseudomonas aeruginosa and Staphylococcus aureus as model pathogens. Correlated chemical imaging will be used to define the onset and range of intercellular quorum sensing signaling in space and time. The biosecretome and interactions of specific species in co-culture biofilms will be characterized for both intra- and inter-species interactions. Also, an ex vivo lung assay will be created to study the spatial biofilm secretome. This research will exploit the ability to spatially map specific chemical products produced by these and other pathogenic bacteria thereby yielding deep and therapeutically informative insights into host colonization, infection, and virulence.

Public Health Relevance

Most bacterial infections are the result of biofilm communities that attach to and colonize host surfaces. We propose to extend the multiplex analysis approach based on correlated mass spectrometric and Raman imaging tools developed within the current R01AI113219 grant to study host-associated co-culture microbial communities growing in situ on surfaces. We will use our methodology to define the onset and range of intercellular quorum sensing signaling in space and time, characterize the biosecretome and interaction of specific species in co-culture biofilms, and recapitulate microbial colonization in an ex vivo lung.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI113219-07
Application #
10021393
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Brown, Liliana L
Project Start
2014-06-01
Project End
2024-08-31
Budget Start
2020-09-01
Budget End
2021-08-31
Support Year
7
Fiscal Year
2020
Total Cost
Indirect Cost
Name
University of Notre Dame
Department
Engineering (All Types)
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
824910376
City
Notre Dame
State
IN
Country
United States
Zip Code
46556
Mattingly, Anne E; Weaver, Abigail A; Dimkovikj, Aleksandar et al. (2018) Assessing Travel Conditions: Environmental and Host Influences On Bacterial Surface Motility. J Bacteriol :
Dunham, Sage J B; Ellis, Joseph F; Baig, Nameera F et al. (2018) Quantitative SIMS Imaging of Agar-Based Microbial Communities. Anal Chem 90:5654-5663
Tomoshige, Shusuke; Dik, David A; Akabane-Nakata, Masaaki et al. (2018) Total Syntheses of Bulgecins A, B, and C and Their Bactericidal Potentiation of the ?-Lactam Antibiotics. ACS Infect Dis 4:860-867
Hu, Jiayun; Fu, Kaiyu; Bohn, Paul W (2018) Whole-Cell Pseudomonas aeruginosa Localized Surface Plasmon Resonance Aptasensor. Anal Chem 90:2326-2332
Morales-Soto, Nydia; Dunham, Sage J B; Baig, Nameera F et al. (2018) Spatially dependent alkyl quinolone signaling responses to antibiotics in Pseudomonas aeruginosa swarms. J Biol Chem 293:9544-9552
Si, Tong; Li, Bin; Comi, Troy J et al. (2017) Profiling of Microbial Colonies for High-Throughput Engineering of Multistep Enzymatic Reactions via Optically Guided Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. J Am Chem Soc 139:12466-12473
Dunham, Sage J B; Ellis, Joseph F; Li, Bin et al. (2017) Mass Spectrometry Imaging of Complex Microbial Communities. Acc Chem Res 50:96-104
Hu, Jiayun; Bohn, Paul W (2017) Optical Biosensing of Bacteria and Bacterial Communities. J Anal Test 1:
Polisetti, Sneha; Baig, Nameera F; Morales-Soto, Nydia et al. (2017) Spatial Mapping of Pyocyanin in Pseudomonas Aeruginosa Bacterial Communities Using Surface Enhanced Raman Scattering. Appl Spectrosc 71:215-223
Si, Tong; Li, Bin; Zhang, Ke et al. (2016) Characterization of Bacillus subtilis Colony Biofilms via Mass Spectrometry and Fluorescence Imaging. J Proteome Res 15:1955-62

Showing the most recent 10 out of 16 publications