Development of a zebrafish sepsis model for high-throughput drug discovery
Anju Mary Philip1,2,3, Youdong Wang1,2, Antonio Mauro1,2,4,5
1Zebrafish Centre for Advanced Drug Discovery, St. Michael's Hospital, 209 Victoria St, Toronto, Ontario, Canada M5B 1T8.
Abstract:
Sepsis is a leading cause of death worldwide. Current treatment modalities remain largely supportive. Intervention strategies focused on inhibiting specific mediators of the inflammatory host response have been largely unsuccessful, a consequence of an inadequate understanding of the complexity and heterogeneity of the innate immune response. Moreover, the conventional drug development pipeline is time consuming and expensive and the low success rates associated with cell-based screens underline the need for whole organism screening strategies, especially for complex pathological processes. Here, we established an LPS-induced zebrafish endotoxemia model, which exhibits the major hallmarks of human sepsis including, edema and tissue/organ damage, increased vascular permeability and vascular leakage accompanied by an altered expression of cellular junction proteins, increased cytokine expression, immune cell activation and ROS production, reduced circulation and increased platelet aggregation. We tested the suitability of the model for phenotype-based drug screening using three primary readouts: mortality, vascular leakage, and ROS production. Preliminary screening identified fasudil, a drug known to protect against vascular leakage in murine models, as a lead hit thereby validating the utility of our model for sepsis drug screens. This zebrafish sepsis model has the potential to rapidly analyze sepsis associated pathologies and cellular processes in the whole organism, as well as to screen and validate large numbers of compounds that can modify sepsis pathology in vivo.
Insights
Developing a zebrafish sepsis model offers a rapid, whole-organism approach for drug discovery. This model successfully identified fasudil, demonstrating its potential for screening sepsis treatments and understanding complex immune responses.
Area of Science:
- * Infectious disease research
- * Innate immunity and inflammation
- * Zebrafish as a model organism
Background:
- * Sepsis is a critical global health issue with limited effective treatments.
- * Current sepsis therapies are primarily supportive, with targeted interventions often failing due to immune response complexity.
- * Traditional drug development is slow, expensive, and has low success rates, necessitating novel screening strategies.
Purpose of the Study:
- * To establish and validate a lipopolysaccharide (LPS)-induced zebrafish endotoxemia model for sepsis research.
- * To assess the model's suitability for high-throughput, phenotype-based drug screening.
- * To identify potential therapeutic compounds for sepsis using this whole-organism model.
Main Methods:
- * Establishment of an LPS-induced zebrafish endotoxemia model mimicking human sepsis hallmarks.
- * Evaluation of the model using mortality, vascular leakage, and reactive oxygen species (ROS) production as primary readouts.
- * Preliminary drug screening to identify compounds modulating sepsis pathology.
Main Results:
- * The zebrafish model recapitulated key sepsis features: edema, tissue damage, increased vascular permeability, altered cellular junctions, cytokine expression, immune cell activation, ROS production, reduced circulation, and platelet aggregation.
- * The model demonstrated efficacy in identifying fasudil, a known vascular leakage inhibitor, as a lead compound.
- * Fasudil's identification validated the model's utility for identifying sepsis therapeutics.
Conclusions:
- * The LPS-induced zebrafish model provides a rapid, cost-effective platform for studying sepsis pathophysiology in vivo.
- * This model is suitable for phenotype-based drug screening, accelerating the identification of novel sepsis treatments.
- * The zebrafish model holds significant potential for advancing sepsis research and therapeutic development.
