A Fluorescence-Based High-Throughput Screening Assay to Identify Growth Inhibitors of the Pathogenic Fungus
Thomas M Smith1, Daryl L Richie2, Jianshi Tao3
1Center for Proteomic Chemistry, Novartis Institutes for BioMedical Research, Inc., 250 Massachusetts Avenue, Cambridge, MA, 02139, USA. thomasm.smith@novartis.com.
Abstract:
Due to the advancements in modern medicine that have resulted in an increased number of immunocompromised individuals, the incidences and the associated mortality of invasive aspergillosis have continued to rise over the past three decades despite appropriate treatment. As a result, invasive aspergillosis has emerged as a leading cause of infection-related mortality in immunocompromised individuals. Utilizing the resazurin to resorufin conversion fluorescence readout to monitor cell viability, herein, we outline a high-throughput screening method amenable to profiling a large pharmaceutical library against the clinically relevant but less frequently screened fungal pathogen Aspergillus fumigatus. This enables the user to conduct high-throughput screening using a disease-relevant fungal growth assay and identify novel antifungal chemotypes as drug leads.
Insights
Invasive aspergillosis is a growing threat in immunocompromised patients. This study presents a new high-throughput screening method to discover novel antifungal drugs against Aspergillus fumigatus.
Area of Science:
- Medical Mycology
- Drug Discovery
- Infectious Diseases
Background:
- Invasive aspergillosis incidence and mortality are rising, particularly in immunocompromised populations, despite current treatments.
- Aspergillus fumigatus is a significant cause of infection-related mortality in these patients.
- There is a critical need for novel antifungal therapies.
Purpose of the Study:
- To develop and validate a high-throughput screening (HTS) assay for identifying new antifungal compounds.
- To profile a large pharmaceutical library against Aspergillus fumigatus using the developed HTS method.
- To identify novel antifungal chemotypes as potential drug leads for invasive aspergillosis.
Main Methods:
- Development of a cell viability assay using resazurin to resorufin conversion fluorescence.
- Implementation of a high-throughput screening method for profiling pharmaceutical libraries.
- Utilizing a disease-relevant fungal growth assay against Aspergillus fumigatus.
Main Results:
- A robust and scalable HTS method was established for Aspergillus fumigatus screening.
- The assay successfully monitored fungal growth and cell viability.
- The method is amenable to screening large compound libraries for antifungal activity.
Conclusions:
- The developed HTS assay provides a valuable tool for discovering novel antifungal agents against Aspergillus fumigatus.
- This approach can accelerate the identification of new drug leads to combat invasive aspergillosis.
- The method facilitates research into less frequently screened fungal pathogens.
