Our hypothesis contends that manufactured carbon nanoparticles (fullerenes, single-walled nanotubes, and multi-walled nanotubes), when exposed to barrier epithelial (bronchiolar, intestinal, and kidney) cells, exert biological effects directed at the membrane and associated cytoskeletal proteins that alter cell function. Specifically, vesicular trafficking and the cytoskeletal components and events associated with vesicular trafficking are likely targets. Alterations in membrane-associated proteins will also alter or dysregulate electrogenic transport mechanisms. We propose that these effects are directly related to the size and physico-chemical character of the carbon nanoparticles, and, at some specific dose, are likely to be injurious to the cells. They thus may pose a significant health hazard. To study this, we plan to expose the three cell types to the three carbon nanomaterials mentioned above, in vitro, using a dose-response design spanning 3 orders of magnitude for a duration of 24 and 48 hrs. After exposure, we will examine differential protein expression using quantitative mass spectrometry and two-dimensional electrophoresis. We will also characterize alterations in protein post-translatiohal modification using those proteomic techniques. We will examine the functional effects of the exposures by measuring ion transport and transepithelial electrical resistance and the potential changes induced by nanoparticle exposure for the response to normal, endogenous regulators. In addition, we will quantify irritation/injury/toxicity using various measurements of cell cytokine secretion, reactive-oxygen species formation, and cell viability. An important aspect of this project is to accurately assess the elemental composition, particle number, particle size distribution, particle shape, and surface area of the primary particle and agglomerated material, as well as the culture-media agglomeration/de-agglomeration status. The results of these studies will provide important new information regarding the effect that different carbon nanoparticles have on barrier epithelial cell global protein expression. In addition, the studies will correlate the changes in protein expression with changes in cellular function at both toxic and subtoxic levels.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Special Emphasis Panel (ZES1-SET-G (NT))
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Lewis, Catherine D
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Indiana University-Purdue University at Indianapolis
Schools of Medicine
United States
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Tilton, Susan C; Karin, Norman J; Tolic, Ana et al. (2014) Three human cell types respond to multi-walled carbon nanotubes and titanium dioxide nanobelts with cell-specific transcriptomic and proteomic expression patterns. Nanotoxicology 8:533-48
Banga, Amiraj; Flaig, Stephanie; Lewis, Shanta et al. (2014) Epinephrine stimulation of anion secretion in the Calu-3 serous cell model. Am J Physiol Lung Cell Mol Physiol 306:L937-46
Lai, Xianyin (2013) Reproducible method to enrich membrane proteins with high purity and high yield for an LC-MS/MS approach in quantitative membrane proteomics. Electrophoresis 34:809-17
Bergin, Ingrid L; Witzmann, Frank A (2013) Nanoparticle toxicity by the gastrointestinal route: evidence and knowledge gaps. Int J Biomed Nanosci Nanotechnol 3:
Lai, Xianyin; Blazer-Yost, Bonnie L; Clack, James W et al. (2013) Protein expression profiles of intestinal epithelial co-cultures: effect of functionalised carbon nanotube exposure. Int J Biomed Nanosci Nanotechnol 3:
Hunter, Chad S; Malik, Raleigh E; Witzmann, Frank A et al. (2013) LHX3 interacts with inhibitor of histone acetyltransferase complex subunits LANP and TAF-1* to modulate pituitary gene regulation. PLoS One 8:e68898
Shannahan, Jonathan H; Brown, Jared M; Chen, Ran et al. (2013) Comparison of nanotube-protein corona composition in cell culture media. Small 9:2171-81
Lai, Xianyin; Agarwal, Mangilal; Lvov, Yuri M et al. (2013) Proteomic profiling of halloysite clay nanotube exposure in intestinal cell co-culture. J Appl Toxicol :
Prince, Kelly L; Colvin, Stephanie C; Park, Soyoung et al. (2013) Developmental analysis and influence of genetic background on the Lhx3 W227ter mouse model of combined pituitary hormone deficiency disease. Endocrinology 154:738-48
Lai, Xianyin; Wang, Lianshui; Witzmann, Frank A (2013) Issues and applications in label-free quantitative mass spectrometry. Int J Proteomics 2013:756039

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