The Anaerobic Rose Chamber Imaging Method (ARCIM) for Probing the Live C. difficile Dynamics
Document Type
Abstract
Publication Date
11-7-2024
DOI
10.25776/2p5j-9m24
Abstract
Clostridioides difficile infection (CDI) is caused by an intestinal, toxin-producing, biofilm-forming obligate anaerobe that is resistant to most broad-spectrum antibiotics. CDI presents an urgent public health threat due to high recurrence rates and limited treatment options, affecting up to 500,000 Americans annually [1]. Although progress has been made in understanding CDI pathophysiology, the development of effective therapies is hindered by the lack of accessible experimental platforms compatible with the obligate anaerobe physiology. While in vivo models have been instrumental in elucidating CDI mechanisms and advancing treatment protocols, their wide-spread use is limited by costs, availability, ethical considerations, and regulatory controls. In contrast, the in vitro microbe-microbe CDI models offer accessibility, precise experimental control, and allow the analysis of microbial interactions, but are limited by the lack of host representation. Furthermore, the development of accessible in vitro host-microbe platforms is challenged by the conflicting oxygen requirements of mammalian cells and anaerobic microbes, creating a critical gap in early CDI modeling.
This work introduces novel applications of the Anaerobic Rose Chamber Imaging Method (ARCIM), which enables the host-microbe co-culture in optically clear, gas- and liquid-tight anaerobic environment. Validation of the ARCIM platform is demonstrated by visualizing toxin-mediated cytoskeletal disruption in mammalian monolayers, establishing the effectiveness of this approach for capturing the real-time host-pathogen interactions. Additionally, the use of a novel oxygen-independent, far-red pTet-IFP2.0 C. difficile reporter is used for dynamic visualization of live biofilm formation. Therefore, ARCIM addresses a significant gap in C. difficile research by enabling accessible, real-time imaging of the host-pathogen interface. Additionally, successful visualization of live biofilm dynamics is paving the way to previously unavailable characterization for spatiotemporal studies of transcriptional regulation within biofilms.
1. Centers for Disease Control and Prevention. C. diff (Clostridioides difficile). Available online: https://www.cdc.gov/c-diff/index.html (accessed on August 31, 2024).
Repository Citation
Zvonareva, Tatiana; Madril, Melissa; Scott, Cody; Zivla, Eleni; Courson, David; and Purcell, Erin, "The Anaerobic Rose Chamber Imaging Method (ARCIM) for Probing the Live C. difficile Dynamics" (2024). 2024 Frank Reidy Research Center for Bioelectrics Retreat. 11.
https://digitalcommons.odu.edu/bioelectrics-2024retreat/11
