High Throughput Methods

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High-throughput technologies such as CRISPR screens, single-cell RNA sequencing, and multi-omics integration enable rapid identification and validation of drug targets, disease mechanisms, and toxicity pathways. This section highlights how omics-based NAMs are reshaping mechanistic understanding and accelerating the discovery of novel therapies for diseases with high unmet medical needs.

Tox21 Consortium: High-Throughput Chemical Screening

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The Tox21 federal collaboration uses robotic high-throughput screening to evaluate thousands of chemicals simultaneously, leading to the development of an 18-assay battery for the estrogen receptor pathway. The EPA formally accepted this computational model as an alternative to traditional rodent assays for identifying endocrine disruptors. This marked the first regulatory prioritization of robotically derived molecular data over animal testing.
Tox21: Chemical testing in the 21st century
United States Federal Government TOX21 Collaboration
About Tox21

Multi-Omics Integration for Toxicity Pathways

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Integrative NAMs combining genomics, transcriptomics, proteomics, and metabolomics have revealed novel oxidative stress and mitochondrial dysfunction signatures. These models successfully distinguished adaptive from adverse responses and generated candidate biomarkers for early detection. This has reshaped the mechanistic understanding of chemical toxicity by moving beyond simple observational data.
Multi-omics integration analysis
The future of pharmaceuticals: Artificial intelligence in drug discovery and development
Skin Sensitisation Case Study
Prospects and challenges of multi-omics data integration in toxicology

CRISPR Screens for Drug Target Validation

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CRISPR/Cas9 gene editing and single-cell RNA sequencing enabled the rapid identification and validation of drug targets for cancer and neurodegenerative diseases. This link between gene perturbations and therapeutic efficacy was validated across multiple cell lines, accelerating the discovery of novel treatments. Consequently, reliance on animal models for target validation has been significantly reduced.
CRISPR Cas9 Gene Editing
CRISPR-Cas9 in Functional Genomics: Implications for Target Validation in Precision Oncology
Target Validation with CRISPR
The future of pharmaceuticals: Artificial intelligence in drug discovery and development

AOP-Linked In Vitro Screens for Seizure Liability

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A government-industry collaboration mapped mechanisms leading to drug-induced seizures using adverse outcome pathways and in vitro assays. The project identified 27 biological target families and developed over 100 assay endpoints for more accurate risk assessment. This mechanism-focused screening replaces animal models that historically failed to predict drug-induced seizures.
De-risking seizure liability
Can New Approach Methodologies De-Risk Drug Development?
iPSC derived cardiomyocytes for cardiac toxicity assessment