An interactive discussion at the public symposium “Using New Approach Methodologies to Address Variability and Susceptibility Across Populations”, explored how nonanimal testing methods might help protect human populations at increased risk of harm from chemical exposures. The diverse set of stakeholders in attendance stressed that activities to characterise chemical risk should be designed to protect all populations, especially the most vulnerable. Such characterisations should also address cumulative exposure from both chemical and nonchemical stressors.