Next generation risk assessment of biocides (PHMG-p and CMIT/MIT)-induced pulmonary fibrosis using adverse outcome pathway-based transcriptome analysis

Next-generation risk assessment (NGRA) has emerged as a promising alternative to non-animal studies owing to the increasing demand for the risk assessment of inhaled toxicants. In this study, NGRA was used to assess the inhalation risks of two biocides commonly used as humidifier disinfectants: polyhexamethylene guanidine phosphate (PHMG-p) and chloromethylisothiazolinone/methylisothiazolinone (CMIT/MIT). Human bronchial epithelial cell transcriptomic data were processed based on adverse outcome pathways and used to establish transcriptome-based points of departure (tPODs) for each biocide. tPOD values were 0.00500–0.0510 μg/cm2 and 0.0342–0.0544 μg/cm2 for PHMG-p and CMIT/MIT, respectively. tPODs may provide predictive power comparable to that of traditional animal-based PODs (aPODs). The tPOD-based NGRA determined that both PHMG-p and CMIT/MIT present a high inhalation risk. Moreover, the identified PHMG-p posed a higher risk than CMIT/MIT, and children were identified as more susceptible population compared to adults. This finding is consistent with observations from actual exposure events. Our findings suggest that NGRA with transcriptomics offers a reliable approach for risk assessment of specific humidifier disinfectant biocides, while acknowledging the limitations of current models and in vitro systems, particularly regarding uncertainties in pharmacokinetics (PK) and pharmacodynamics (PD). Our study utilizes Next-generation Risk Assessment (NGRA) to evaluate the inhalation risks of PHMG-p and CMIT/MIT, common biocides in humidifier disinfectants. Transcriptomic data from human bronchial epithelial cells were employed to establish transcriptome-based points of departure (tPODs), revealing high inhalation risks for both biocides, with PHMG-p posing a higher risk than CMIT/MIT. Children were identified as a more susceptible population, consistent with real-world exposure events. This research identifies these biocides as hazardous due to their potential respiratory health impacts and provides a more reliable risk assessment approach using NGRA with transcriptomics, contributing to addressing environmental concerns over biocide exposure.