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Diagnosing Lung Cancer in Exhaled Breath using Gold Nanoparticles

Reporter-curator: Tilda Barliya PhD

Authors: Gang Peng, Ulrike Tisch, Orna Adams¹, Meggie Hakim, Nisrean Shehada, Yoav Y. Broza, Salem Billan, Roxolyana Abdah-Bortnyak, Abraham Kuten & Hossam Haick. (NATURE NANOTECHNOLOGY | VOL 4 | OCTOBER 2009 |)

Abstract:

Conventional diagnostic methods for lung cancer1,2 are unsuitable for widespread screening, because they are expensive and occasionally miss tumours. Gas chromatography/mass spectrometry studies have shown that several volatile organic compounds, which normally appear at levels of 1–20 ppb in healthy human breath, are elevated to levels between 10 and 100 ppb in lung cancer patients. Here we show that an array of sensors based on gold nanoparticles can rapidly distinguish the breath of lung cancer patients from the breath of healthy individuals in an atmosphere of high humidity. In combination with solidphase microextraction, gas chromatography/mass spectrometry was used to identify 42 volatile organic compounds that represent lung cancer biomarkers. Four of these were used to train and optimize the sensors, demonstrating good agreement between patient and simulated breath samples. Our results show that sensors based on gold nanoparticles could form the basis of an inexpensive and non-invasive diagnostic tool for lung cancer. (http://www.nature.com/nnano/journal/v4/n10/abs/nnano.2009.235.html) (lnbd.technion.ac.il/NanoChemistry/SendFile.asp?DBID=1…1…) 

Introduction:

Lung cancer accounts for 28% of cancer-related deaths. Approximately 1.3 million people die worldwide every year. Breath testing is a fast, non-invasive diagnostic method that links specific volatile organic compounds (VOCs) in exhaled breath to medical conditions. Gas chromatography/mass spectrometry (GC-MS), ion flow tube mass spectrometry10, laser absorption spectrometry,infrared spectroscopy, polymer-coated surface acoustic wave sensors and coated quartz crystal microbalance sensors have been used for this purpose. However, these techniques are expensive, slow, require complex instruments and, furthermore, require pre-concentration of the biomarkers (that is, treating the biomarkers by a process to increase the relative concentration of the biomarkers to a level that can be detected by the specific technique) to improve detection.

Here, we report a simple, inexpensive, portable sensing technology to distinguish the breath of lung cancer patients from healthy subjects without the need to pre-treat the exhaled breath in any way (see also refs 14–16 for the diagnosis of lung cancer by sensing technology that is based on arrays of polymer/carbon black sensors). Our study consisted of four phases and included volunteers aged 28–60 years. Samples were collected from 56 healthy controls and 40 lung cancer patients after clinical diagnosis using conventional methods and before chemotherapy or other treatment.

In the first phase, we collected exhaled alveolar breath of lung cancer patients and healthy subjects using an ‘offline’ method. This method was designed to avoid potential errors arising from the failure to distinguish endogenous compounds from exogenous ones in the breath and to exclude nasal entrainment of the gas. Exogenous VOCs can be either directly absorbed through the lung via the inhaled breath or indirectly through the blood or skin. Endogenous VOCs are generated by cellular biochemical processes in the body and may provide insight into the body’s function

In the second phase, we identified the VOCs that can serve as biomarkers for lung cancer in the breath samples and determined their relative compositions, using GC-MS in combination with solidphase microextraction (SPME). GC-MS analysis identified over 300–400 different VOCs per breath sample, with .87% reproducibility for a specific volunteer examined multiple times over a period of six months. Forward stepwise discriminant analysis identified 33 common VOCs that appear in at least 83% of the patients but in fewer than 83% of the healthy subjects

The compounds that were observed in both healthy breath and lung cancer breath were presented not only at different concentrations but also in distinctively different mixture compositions.

Further forward stepwise discriminant analysis revealed nine uncommon VOCs that appear in at least 83% of the patients but not in the majority (83%) of healthy subjects. This additional class of VOCs has not been recognized in earlier GC-MS studies.

In spite of these advances in the GC-MS analysis, these data certainly do not account for all the VOCs present in the exhaled breath samples, because the pre-concentration technique can be thought of as a solid phase that extracts only part of the analytes present in the examined phase and, subsequently, releases only part of the extracted analytes.

So, it is likely that the actual mixture of VOCs to which, for example, an array of gold nanoparticle sensors would be responding  is different from that obtained by GC-MS.

In the third phase of this study we designed an array of nine crossreactive chemiresistors, in which each sensor was widely responsive to a variety of odorants for the detection of lung cancer by means of breath testing. We used chemiresistors based on assemblies of 5-nm gold nanoparticles  with different organic functionalities (dodecanethiol, decanethiol, 1-butanethiol, 2-ethylhexanethiol, hexanethiol, tert-dodecanethiol, 4-methoxy-toluenethiol, 2-mercaptobenzoxazole and 11-mercapto-1-undecanol).

Chemiresistors based on functionalized gold nanoparticles combine the advantages of organic specificity with the robustness and processability of inorganic materials.

The response of the nine-sensor array to both healthy and lung cancer breath samples was analysed using principal component analysis . It can be seen that there is no overlap of the lung cancer and healthy patterns.

The PCA of the healthy control group revealed that the set of gold nanoparticles sensors was not influenced by characteristics such as gender, age or smoking habits, thus strengthening the ability of the sensors to discriminate between healthy and cancerous breath. Experiments with a wider population of volunteers to thoroughly probe the influence of diet, alcohol consumption,metabolic state and genetics are under way and will be published elsewhere.

Summary:

To summarize, we have demonstrated that an array of chemiresistors based on functionalized gold nanoparticles in combination with pattern recognition methods can distinguish between the breath of lung cancer patients and healthy controls, without the need for dehumidification or pre-concentration of the lung cancer biomarkers. Our results show great promise for fast, easy and cost-effective diagnosis and screening of lung cancer. The developed devices are expected to be relatively inexpensive, portable and amenable to use in widespread screening, making them potentially valuable in saving millions of lives every year. Given the impact of the rising incidence of cancer on health budgets worldwide, the proposed technology will be a significant saving for both private and public health expenditures. The potential exists for using the proposed technology to diagnose other conditions and diseases, which could mean additional cost reductions and enhanced opportunities to save lives.

Ref:

1. Gang Peng, Ulrike Tisch, Orna Adams, Meggie Hakim, Nisrean Shehada, Yoav Y. Broza, Salem Billan, Roxolyana Abdah-Bortnyak, Abraham Kuten& Hossam Haick. Diagnosing lung cancer in exhaled breath using gold nanoparticles. Nature Nanotechnology 4, 669 – 673 (2009) http://www.nature.com/nnano/journal/v4/n10/abs/nnano.2009.235.html

2. http://lungcancer.about.com/od/diagnosisoflungcancer/a/diagnosislungca.htm

3. http://metabolomx.com/2011/12/15/metabolomx-test-detects-lung-cancer-from-breath/

4. http://www.chestnet.org/accp/pccsu/medical-applications-exhaled-breath-analysis-and-testing?page=0,3