93 Integrated analysis of cellular pathways of respiratory nanotoxicity

Abstract Nanomaterials (NMs) offer wide applications but raise concerns about respiratory toxicity with potential contributions to the development of chronic lung diseases. Therefore, it is imperative to understand the mechanism underlying the respiratory toxicity of nanomaterials. While numerous animal- and cell-based studies have been applied to investigate the respiratory toxicity of various nanomaterials, and transcriptomic profiling (microarray, RNA-seq) has generated extensive public gene expression data, bulk datasets lack cell-type resolution and cannot reveal cell-specific responses. Our recent single-cell transcriptomic mouse study (Voss, Han et al., ACS Nano, 2025), revealed material-specific pulmonary perturbations and distinct inflammatory responses induced by different NMs: spherical and soot-like carbon nanoparticles (CNP), flexible double-walled carbon nanotubes (DWCNT), and rigid multi-walled carbon nanotubes (MWCNT). However, single-cell analysis is costly and currently available only for a limited number of materials. To address this, we propose an approach that integrates public bulk RNA-seq datasets with existing single-cell RNA-seq references to deconvolute cell-type-specific responses across a broader range of NMs. This approach aims to identify key responsive cell types, top-regulated genes, and early toxicity pathways. We have collected bulk datasets related to the respiratory toxicity of nanomaterials by mining public databases. We are currently applying and comparing different deconvolution methods (eg MuSiC and CIBERSORTx) to bulk data, using our lab-generated single-cell sequencing data as a reference. Next steps include summarising material-specific cell proportion changes, biomarkers or pathways. This will improve our understanding enable the development of in vitro models to simulate these perturbations, supporting future animal-free toxicity testing and contributing to the safe-by-design development of future nanomaterials.