Of blood and breath: metabolite-based diagnosis of ovarian cancer

Physicians always knew that breath contains clues to diseases. Chemicals in breath often correlate with chemicals in saliva and blood - be it alcohol, anaesthetics or other metabolites (see, for example, this study by Dr Andreas Hengstenberg).

As one of my interests is breath-based detection of ovarian cancer, I took note of the recent paper claiming 99% to 100% accuracy of detecting ovarian cancer by metabolites in blood.
The authors used customized functional support vector machine-based machine-learning algorithms to classify thousands of metabolites measured by mass spectrometry (JEOL AccuTOF™ DART® that allowed to forego conventional liquid chromatography as sufficient resolution was achieved without separation) in peripheral blood. 

100% sensitivity and 100% specificity was achieved with 64-30 split validation technique, while 100% sensitivity and 98% specificity was the accuracy of leave-one-out-cross-validation. Very large number of metabolites, from 2,000 to 3,000 features, contributed to such discriminatory power (see the list of 14,000+ in supplemental material) 
 

Set of 25 canonical metabolic pathways relevant to the uploaded elemental
formulae ranked according to their p-values (hypergeometric distribution).

Histamine, amino acid, fructose and glucose metabolism were among the most prominent processes discriminating cancer and healthy blood.
It's that simple: sugar feeds cancer. Scientists have long found that cancer cells slurp fructose, and that fructose intake can be linked to some cancers. Histamine/polyamine interplay in cancers is also known. Histamine may be involved in inhibition of the local immune response against cancer. Is amino acid metabolism also linked to cancer? Well, what is not.   




Metabolomic biomarkers were always known to have diagnostic potential - cholesterol and glucose are among the oldest and most widely performed diagnostic tests. Yet, most bleeding edge cancer detection platforms are genomic or proteomic in nature.  Of the thousands of known biomarkers, only a handful have made it into the clinic. Existing ovarian cancer tests mostly rely on detecting a protein -  carbohydrate antigen 125. Vermillion's OVA1 and HealthLinx OvPlex tests use five proteins. This may be extended to 7.

Metabolites represent the end products of the genome and proteome, thus metabolomics-based diagnostics  holds the promise of providing powerful diagnostics,  allowing for differentiation of increased and decreased levels of chemicals with low process coefficient of variation.


Metabolomic tests were used for medical diagnostics starting with Hippocrates and Lavoisier. They continue to be explored by modern scientists. Dr Michael Phillips, for example, developed HeartsBreath Test, approved by the US Food and Drug Agency for early diagnosis of heart transplant rejection. Research proved the potential of inexpensive breath tests in discriminating lung, breast, colon and prostate cancers. Let's hope the new article  - along with others - will lead to novel consumer products, not only more academic research and peer-reviewed publications.



Zhou M, Guan W, Walker LD, Mezencev R, Benigno BB, Gray A, Fernández FM, & McDonald JF (2010). Rapid Mass Spectrometric Metabolic Profiling of Blood Sera Detects Ovarian Cancer with High Accuracy. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology PMID: 20699376

On cancers and petroleum spills

Researchers have known for years that smell of cancer patients is chemically different from healthy individuals. One more study featured in British Journal of Cancer brings us a bit closer to an inexpensive, easy-to-use, portable device for home diagnostics. 

Exhaled breath collected from 177 volunteers (patients with lung, colon, breast, and prostate cancers and healthy controls) was examined by gold nanoparticle nanosensor arrays (GNPs) and gas chromatography linked to the mass spectrometry technique (GC-MS). 

GNP sensor resistance responses showed remarkable separation between cancer and healthy controls (Principal Component Analysis results are shown in the Figure: LC, lung cancer;  CC, colon cancer; BC, breast cancer; PC, prostate cancer).

Most of the VOCs reported in this study appear for the first time in the literature, adding to the wide spectrum of chemicals previously proposed as cancer biomarkers. Some of the chemicals -  predictive of  lung and prostate cancers -  are frequently released to the environment through petroleum spills. 

1-methyl-4-(1-methylethyl)-benzene - known as p-cymene  or p-isopropyltoluene -   is utilized by various species for chemical communication. It can be derived from the essential oils of herbs and spices and has biocidal properties against foodborne pathogens such as spoilage yeasts and E. coli O157:H7. p-cymene is the biological precursor of carvacrol that is also an antimicrobial agent (Kiskó and Roller, 2005).  It's decreased with cancer, and is present at higher concentrations in healthy individuals.


Toluene, dodecane and other aromatic components of petroleum are among chemicals found in human breath.

Typical "octane booster" toluene  - present at higher concentrations in lung and prostate cancers - is toxic to living organisms although some bacteria (like P. putida that has toluene operon) are able to grow in its presence (Eaton 1997).

Dodecane, a biogasoline component, is higher in the breath of healthy individuals. It is decreased in lung cancer. One of its derivatives - 2,6,11-trimethyl-dodecane was found in 80% of the males, but in none of the females participating in the study. 

Another aromatic compound, 2-amino-5-isopropyl-8-methyl-1-azulenecarbonitrile, similar to carbonitriles used in manufacturing of fragrance agents, is present at higher concentrations in breast, colon and prostate cancers when compared to healthy controls.

An alcane 3,7-dimethyl-undecane was found to be indicative of allergies. It was previously found to be eliminated from mice odors when they enter reproductive cycle (Achiraman & Archunan, 2006) and proposed to be used in diagnostics of asthma (Dragonieri et al., 2007).

Chemicals in breath can tell not only about cancers, but also relate to other diseases, environmental exposures and dietary behavior. This might decrease discriminative power of  expensive metabolomics technologies and bioinformatics approaches not based on additional knowledge, yet custom-made sensor arrays show great promise.


References


Peng G, Hakim M, Broza YY, Billan S, Abdah-Bortnyak R, Kuten A, Tisch U, & Haick H (2010). Detection of lung, breast, colorectal, and prostate cancers from exhaled breath using a single array of nanosensors. British journal of cancer, 103 (4), 542-51 PMID: 20648015



Gabriella Kiskó, Sibel Roller. (2005) Carvacrol and p-cymene inactivate Escherichia coli O157:H7 in apple juice.  BMC Microbiol. 2005; 5: 36

Eaton RW.  (1997) p-Cymene catabolic pathway in Pseudomonas putida F1: cloning and characterization of DNA encoding conversion of p-cymene to p-cumate. J Bacteriol. 1997 May;179(10):3171-80.

Shanmugam Achiraman, Govindaraju Archunan (2006) 1-Iodo-2methylundecane, a putative estrus-specific urinary chemo-signal of female mouse (Mus musculus) Theriogenology 66, 1913–1920

Dragonieri S, Schot R, Mertens BJ, Le Cessie S, Gauw SA, Spanevello A, Resta O, Willard NP, Vink TJ, Rabe KF, Bel EH, Sterk PJ. (2007) An electronic nose in the discrimination of patients with asthma and controls. J Allergy Clin Immunol. 120(4):856-62.