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)
Project #
Application #
Study Section
Special Emphasis Panel (ZES1-SET-G (NT))
Program Officer
Lewis, Catherine D
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Indiana University-Purdue University at Indianapolis
Schools of Medicine
United States
Zip Code
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
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 (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
Xia, Tian; Hamilton, Raymond F; Bonner, James C et al. (2013) Interlaboratory evaluation of in vitro cytotoxicity and inflammatory responses to engineered nanomaterials: the NIEHS Nano GO Consortium. Environ Health Perspect 121:683-90
Li, Pin; Lai, Xianyin; Witzmann, Frank A et al. (2013) Bioinformatic Analysis of Differential Protein Expression in Calu-3 Cells Exposed to Carbon Nanotubes. Proteomes 1:219-239
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:
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; Agarwal, Mangilal; Lvov, Yuri M et al. (2013) Proteomic profiling of halloysite clay nanotube exposure in intestinal cell co-culture. J Appl Toxicol :
Lai, Xianyin; Wang, Lianshui; Witzmann, Frank A (2013) Issues and applications in label-free quantitative mass spectrometry. Int J Proteomics 2013:756039

Showing the most recent 10 out of 20 publications