Transforming Preclinical Hepatotoxicity Screening with the I.DOT (Toxicological Sciences)

Assessing the hepatotoxicity of therapeutic small molecule drugs is a critical step in the preclinical drug development process. A recently published study in Toxicological Sciences highlights how I.DOT technology is revolutionizing this crucial area of research.

The study, conducted by researchers, our customers, from InSphero AG in Switzerland, utilized a 384-well microplate system featuring physiological liver microtissues (hLiMTs) to evaluate the cytotoxicity of small molecule drugs. The researchers leveraged the advanced capabilities of the I.DOT Liquid Handler to precisely dispense the DMSO-based drug compounds across the microplate.

Figure 1. The paper "Physiological liver microtissue 384-well microplate system for preclinical hepatotoxicity assessment of therapeutic small molecule drugs" published in Toxicological Sciences.

Preclinical hepatotoxicity assessment of small molecule drugs using the Physiological Liver Microtissue (hLiMT) 384-well microplate system provides a high-throughput, physiologically relevant model to predict liver toxicity. This system employs 3D liver microtissues comprising primary human hepatocytes and supporting liver cells, closely mimicking native liver architecture and function (Messner et al., 2018; Yang et al., 2023).

Given the liver's central role in metabolizing drugs and eliminating toxins, assessing a compound's potential for hepatotoxicity helps predict adverse liver effects in humans (Messner et al., 2018). Therefore, the hLiMT system supports long-term culture, enabling repeated dosing to assess both acute and chronic hepatotoxicity over multiple time points. Assays commonly used include ATP-based luminescence for viability, glutathione (GSH) depletion for oxidative stress, and lactate dehydrogenase (LDH) release for cell membrane integrity (Jaeschke et al., 2002).

Key Advantages of I.DOT for Hepatotoxicity Assessment

The I.DOT technology offers several transformative benefits for preclinical hepatotoxicity screening:

1. Accurate, Low-Volume Dispensing: I.DOT's ability to dispense volumes ranging from nanoliters to microliters with high precision is essential when working with 384-well plates. This minimizes reagent and compound waste, a critical consideration in costly preclinical studies.
2. Reduced Cross-Contamination: The non-contact dispensing of I.DOT eliminates the risk of cross-contamination between wells, preserving the integrity of the liver microtissues and ensuring reliable hepatotoxicity data.
3. Enhanced Throughput: I.DOT's rapid processing of entire 384-well plates significantly accelerates the drug treatment steps, enabling efficient high-throughput screening of multiple compounds, concentrations, and replicates.
4. Improved Sustainability: The efficient use of compounds and reagents in miniaturized volumes by I.DOT helps reduce the costs and environmental impact associated with high-throughput hepatotoxicity studies.

Advancing Preclinical Hepatotoxicity Assessment

The combination of I.DOT's transformative capabilities and the physiologically relevant hLiMT 384-well microplate system represents a powerful approach to preclinical hepatotoxicity screening. This integration allows researchers to:

• Establish accurate dose-response curves and determine IC₅₀ values with confidence
• Assess both acute and chronic hepatotoxicity through repeated dosing
• Explore the effects of drug combinations for combination therapy development

Unlocking the Future of Hepatotoxicity Screening

As the pharmaceutical industry continues to navigate the complexities of small molecule drug development, technologies like I.DOT are poised to become indispensable tools. By enhancing precision, efficiency, and throughput in preclinical hepatotoxicity assessment, I.DOT is transforming the way researchers evaluate the safety and efficacy of therapeutic compounds.

Experience the power of I.DOT technology and discover how it can revolutionize your hepatotoxicity screening workflows. Download the brochure and unlock the future of safer, more sustainable small molecule drug development.

References

Fäs, L., Chen, M., Tong, W., Wenz, F., Hewitt, N.J., Tu, M., Sanchez, K., Zapiórkowska-Blumer, N., Varga, H., Kaczmarska, K. and Colombo, M.V., 2024. Physiological liver microtissue 384-well microplate system for preclinical hepatotoxicity assessment of therapeutic small molecule drugs. Toxicological Sciences, p.kfae123.

Jaeschke, H., Gores, G.J., Cederbaum, A.I., Hinson, J.A., Pessayre, D. and Lemasters, J.J., 2002. Mechanisms of hepatotoxicity. Toxicological sciences, 65(2), pp.166-176.

Messner, S., Fredriksson, L., Lauschke, V.M., Roessger, K., Escher, C., Bober, M., Kelm, J.M., Ingelman-Sundberg, M. and Moritz, W., 2018. Transcriptomic, proteomic, and functional long-term characterization of multicellular three-dimensional human liver microtissues. Applied in vitro toxicology, 4(1), pp.1-12.

Yang, S., Ooka, M., Margolis, R.J. and Xia, M., 2023. Liver three-dimensional cellular models for high-throughput chemical testing. Cell Reports Methods, 3(3).