Direct exposure to inhaled mainstream cigarette smoke is known to cause smoking-related damage in the human lung. Aerosol exposure of human three dimensional (3D) organotypical airway epithelial tissue cultures, growing at an air-liquid interface, is a well-established in vitro model enabling a Systems Biology-based Reduced Risk Product (RRP) assessment. Migration kinetics and metabolism of deposited aerosol compounds are important parameters in understanding their bioavailability. To better understand the role of xenobiotic metabolism after cigarette smoking, human subcellular liver and lung fraction models (microsomes, S9) have been established to assess metabolite profiles and reactive metabolites relevant for toxicological assessment. The integration of exposure characterization results with a systems toxicology approach to measure the biological impact of exposure is informative to the product assessment strategy and has the potential to highlight modes of action associated with RRP exposure.