Impact of confounding by leukocyte composition on associations of leukocyte DNA methylation with common risk factors.

AIM: One concern in epigenome-wide studies investigating leukocyte DNA methylation is that observed associations may at least partly reflect differences in leukocyte composition (LC) rather than changes in methylation. We estimated the magnitude of confounding by LC for common risk factors and diseases. MATERIALS & METHODS: Variation of LC according to sex, race, age, smoking, alcohol consumption, BMI, cardiovascular fitness, hypertension, coronary heart disease and diabetes was analyzed using blood differentials from 4117 participants of NHANES. Furthermore, leukocyte DNA methylation levels of biomarkers of smoking, BMI, diabetes, age and sex were regressed on these outcomes in a sample of 989 participants of ESTHER, and regression coefficients with and without adjustment for estimated LC were compared. RESULTS: Aside from race and ages below 25 years, none of the investigated factors had substantial impact on LC. Adjusted and unadjusted coefficients were virtually identical. CONCLUSION: Confounding by LC might often be a minor issue.

Epigenome-wide discovery and evaluation of leukocyte DNA methylation markers for the detection of colorectal cancer in a screening setting.

BACKGROUND: Colorectal cancer (CRC) is the third most common cancer worldwide. If detected at an early stage, prognosis is good. Despite increasing evidence for the benefits of implemented screening programs, such as screening colonoscopy, compliance is rather low. Hence there is demand for non-invasive tests for the early detection of CRC with high acceptance in population-wide screening. The objective of this study was to identify and evaluate leukocyte DNA methylation patterns as a potential biomarker for early detection of CRC. METHODS: Blood samples of patients scheduled for a screening colonoscopy were collected before the procedure. Additionally, blood samples from CRC cases recruited in a clinical setting were collected. DNA was extracted from leukocytes, and DNA methylation was measured with the Infinium 450K BeadChip. In total, 46 CRC cases and 140 controls from the screening setting and 93 CRC cases from the clinical setting were measured. RESULTS: An epigenome-wide discovery revealed two CpG sites in the promoter region of KIAA1549L that were significantly differentially methylated between cases and controls. A third marker in the body region of BCL2 was discovered in a candidate approach testing biomarkers reported in the literature. Logistic regression models built on these three markers yielded an optimism-corrected c-statistic of 0.69 in the screening setting and 0.73 in the clinical setting. CONCLUSIONS: Although diagnostic performance of the DNA methylation signature identified in this first epigenome-wide association study of leukocyte DNA methylation with CRC in a screening setting is not competitive with established screening tests, the identified markers may contribute to multimarker panels for early detection of CRC.

Detection of cancer through exhaled breath: a systematic review.

BACKGROUND: Timely diagnosis of cancer represents a challenging task; in particular, there is a need for reliable non-invasive screening tools that could achieve high levels of adherence at virtually no risk in population-based screening. In this review, we summarize the current evidence of exhaled breath analysis for cancer detection using standard analysis techniques and electronic nose. METHODS: Relevant studies were identified searching Pubmed and Web of Science databases until April 30, 2015. Information on breath test performance, such as sensitivity and specificity, was extracted together with volatile compounds that were used to discriminate cancer patients from controls. Performance of different breath analysis techniques is provided for various cancers together with information on methodological issues, such as breath sampling protocol and validation of the results. RESULTS: Overall, 73 studies were included, where two-thirds of the studies were conducted on lung cancer. Good discrimination usually required a combination of multiple biomarkers, and area under the receiver operating characteristic curve or accuracy reached levels of 0.9 or higher in multiple studies. In 25% of the reported studies, classification models were built and validated on the same datasets. Huge variability was seen in different aspects among the studies. CONCLUSIONS: Analyses of exhaled breath yielded promising results, although standardization of breath collection, sample storage and data handling remain critical issues. In order to foster breath analysis implementation into practice, larger studies should be implemented in true screening settings, paying particular attention to standardization in breath collection, consideration of covariates, and validation in independent population samples.