title: Nano-risk Science: application of toxicogenomics in an adverse outcome pathway framework for risk assessment of multi-walled carbon nanotubes.
abstract: BACKGROUND
  A diverse class of engineered nanomaterials (ENMs) exhibiting a wide array of physical-chemical properties that are associated with toxicological effects in experimental animals is in commercial use. However, an integrated framework for human health risk assessment (HHRA) of ENMs has yet to be established. Rodent 2-year cancer bioassays, clinical chemistry, and histopathological endpoints are still considered the 'gold standard' for detecting substance-induced toxicity in animal models. However, the use of data derived from alternative toxicological tools, such as genome-wide expression profiling and in vitro high-throughput assays, are gaining acceptance by the regulatory community for hazard identification and for understanding the underlying mode-of-action. Here, we conducted a case study to evaluate the application of global gene expression data in deriving pathway-based points of departure (PODs) for multi-walled carbon nanotube (MWCNT)-induced lung fibrosis, a non-cancer endpoint of regulatory importance.
  
  
  METHODS
  Gene expression profiles from the lungs of mice exposed to three individual MWCNTs with different physical-chemical properties were used within the framework of an adverse outcome pathway (AOP) for lung fibrosis to identify key biological events linking MWCNT exposure to lung fibrosis. Significantly perturbed pathways were categorized along the key events described in the AOP. Benchmark doses (BMDs) were calculated for each perturbed pathway and were used to derive transcriptional BMDs for each MWCNT.
  
  
  RESULTS
  Similar biological pathways were perturbed by the different MWCNT types across the doses and post-exposure time points studied. The pathway BMD values showed a time-dependent trend, with lower BMDs for pathways perturbed at the earlier post-exposure time points (24 h, 3d). The transcriptional BMDs were compared to the apical BMDs derived by the National Institute for Occupational Safety and Health (NIOSH) using alveolar septal thickness and fibrotic lesions endpoints. We found that regardless of the type of MWCNT, the BMD values for pathways associated with fibrosis were 14.0-30.4 μg/mouse, which are comparable to the BMDs derived by NIOSH for MWCNT-induced lung fibrotic lesions (21.0-27.1 μg/mouse).
  
  
  CONCLUSIONS
  The results demonstrate that transcriptomic data can be used to as an effective mechanism-based method to derive acceptable levels of exposure to nanomaterials in product development when epidemiological data are unavailable.
authors.affiliation: Environmental Health Science and Research Bureau, Health Canada, Ottawa, ON, K1A 0K9, Canada.
authors.forename: Sarah
authors.initials: S
authors.lastname: Labib
authors.affiliation: Environmental Health Science and Research Bureau, Health Canada, Ottawa, ON, K1A 0K9, Canada.
authors.forename: Andrew
authors.initials: A
authors.lastname: Williams
authors.affiliation: Environmental Health Science and Research Bureau, Health Canada, Ottawa, ON, K1A 0K9, Canada.
authors.forename: Carole L
authors.initials: CL
authors.lastname: Yauk
authors.affiliation: Environmental Health Science and Research Bureau, Health Canada, Ottawa, ON, K1A 0K9, Canada.
authors.forename: Jake K
authors.initials: JK
authors.lastname: Nikota
authors.affiliation: National Research Centre for the Working Environment, Lerso Parkallé 105, DK-2100, Copenhagen, Denmark.|Department of Public Health, University of Copenhagen, DK-1353, Copenhagen K, Denmark.
authors.forename: Håkan
authors.initials: H
authors.lastname: Wallin
authors.affiliation: National Research Centre for the Working Environment, Lerso Parkallé 105, DK-2100, Copenhagen, Denmark.|Department of Micro- and Nanotechnology, Technical University of Denmark, DK-2800 Kgs, Lyngby, Denmark.
authors.forename: Ulla
authors.initials: U
authors.lastname: Vogel
authors.affiliation: Environmental Health Science and Research Bureau, Health Canada, Ottawa, ON, K1A 0K9, Canada. sabina.halappanavar@hc-sc.gc.ca.
authors.forename: Sabina
authors.initials: S
authors.lastname: Halappanavar
mesh_terms: D000818:Animals; D019985:Benchmarking; D019295:Computational Biology; D030541:Databases, Genetic; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects*; D053263:Gene Regulatory Networks / Q000187:drug effects; D006801:Humans; D008168:Lung / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D036103:Nanotechnology; D037742:Nanotubes, Carbon / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D011658:Pulmonary Fibrosis / Q000139:chemically induced* / Q000235:genetics* / Q000473:pathology; D018570:Risk Assessment; D013997:Time Factors; D043922:Toxicogenetics; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D037742:Nanotubes, Carbon
keywords: Adverse outcome pathways; Benchmark dose; Case study; Lung fibrosis; Nano; Risk assessment; Toxicogenomics