Repository of Amoeba Drug Activity Records 

Summary: This open-source repository provides comprehensive drug susceptibility data for various Acanthamoeba strains, measured using CellTiter-Glo 2.0 (Promega) in PG and Ac6 media. Testing conditions, including seeding density, drug exposure at 27°C for three days, and standardized drug preparation, ensure reliable and comparable data. Despite challenges in harmonizing conditions across the field, a unified approach is essential for advancing drug discovery. This dataset, compiled by the Rice Research Group at Purdue University, aims to support researchers, clinicians, and policymakers in selecting effective treatments and establishing microbial breakpoints for improved patient care from amoebic pathogens.

Project: This data repository was developed in response to the alarming inconsistencies in testing methods, conditions, and strain selection across our field. Variability in growth media, temperature, time-points, DMSO susceptibility, and compound sources-including differences in manufacturers, purity, and batch composition-further complicates drug susceptibility assessments. To ensure these factors do not influence our efficacy data, we have standardized our approach to drug susceptibility testing. Given that many of these compounds are already recommended therapeutics for amoebic diseases, particularly Acanthamoeba keratitis (AK), we are committed to sharing this data with the community.

Here, we optimize the seeding density for all amoeba strains and isolates, ensuring consistent parasite counts per well across various media conditions. For Acanthamoeba, drug exposure is conducted at 27°C for three days, using the CellTiter-Glo 2.0 assay, a well-established method in drug discovery and drug development. To minimize errors in starting concentrations and reduce variability from repeated weighing or freeze-thaw cycles, we prepare drug stocks at 5 mg/ml in neat DMSO, achieving a final screening concentration of 50 µg/ml in 1% DMSO. When working with Acanthamoeba strains more sensitive to low DMSO %’s, we adjust and deviate from our initial protocol by preparing 50 mg/ml stocks to reach the same final screening concentration in 0.1% DMSO. All data is analyzed and stored using CDD Vault, a paid software platform.

The data presented in these tables below represent the mean response from at least three biological replicates per condition and drug. Despite our efforts to standardize conditions, challenges remain-some amoebae only grow in specific media (Ac6, PG, PYG, or M20), at particular temperatures, or require endosymbionts for survival. These variations make harmonization difficult, but a unified approach is critical. If we can agree on consistent strains and conditions across laboratories, we will improve the comparability of drug discovery research in our field.

Current drug discovery standards consider anything above 10 µM as an inadequate starting point for a “hit.” If we hope to develop a targeted anti-amoebic drug, we must align with best practices from other infectious disease research areas.

This project is the first of its kind in our field. We hope that clinicians, researchers, patients, and policymakers can leverage this data to make informed decisions-whether in defining microbial breakpoints or selecting the most effective treatments for specific clinical strains. With continued advancements in diagnostics, personalized treatment plans for amoebic infections may one day become a reality.

Citation and Acknowledgment

You are welcome to incorporate this dataset into your work. If you publish findings or share insights derived from this dataset, we ask that you acknowledge our contribution by citing it as follows:

"RADAR – Repository of Amoeba Drug Activity Records", provided by the Rice Research Group at Purdue University, 2025. Available at: https://vet.purdue.edu/discovery/rice/radar/. 

For publications, presentations, or other works that utilize this dataset, please include the following acknowledgment:

"This study relies on data provided by the Rice Research Group at Purdue University and project RADAR – Repository of Amoeba Drug Activity Records. While we acknowledge their contributions, the authors take full responsibility for all analyses and interpretations presented."

Finally, we would appreciate being informed of any publications or significant applications of this dataset. If you’d like to share your work or have any questions, please reach out to Dr. Christopher A. Rice at carice@purdue.edu.

Download the Dataset