The ALTERNATIVE project aims to revolutionize cardiotoxicity assessments by moving away from traditional animal testing towards a sophisticated platform that integrates advanced microphysiological models, omics technologies, and machine-learning analytics. It provides clear evidence of the environmental chemical impacts on cardiac health, advocating for updates of the toxicological requirements in regulatory frameworks. Through the development of Adverse Outcome Pathway (AOP) networks and a novel Integrated Approach to Testing and Assessment (IATA), ALTERNATIVE provides essential tools to enhance the safety, efficiency and ethical standards of regulatory practices. Moreover, the outcome of the project will support the current European roadmap towards an animal-free regulatory system.