February 9, 2012 at 11:00 am EST
11:00-11:05 Chairperson’s Opening Remarks
11:05-11:30 Profiling of Circulating microRNAs: From Single Biomarkers to Rewired Networks
Manuel Mayr, M.D., Ph.D., Senior Fellow, British Heart Foundation; Professor, Cardiovascular Proteomics, King’s College London
The recent discovery that microRNAs (miRNAs) are present in blood sparked interest in their use as potential biomarkers. These circulating microRNAs are not cell associated but seem to escape degradation by endogenous RNase activity by residing in microvesicles, exosomes, microparticles and apoptotic bodies, although recently the formation of protein-miRNA complexes has also been proposed. While research on circulating miRNAs is still in its infancy, high analytical standards in statistics and study design are a prerequisite to obtain robust data and avoid repeating the mistakes of the early genetic association studies. Otherwise, studies tend to get published because of their novelty despite low numbers, poorly matched cases and controls and no multivariate adjustment for conventional risk factors. Research on circulating miRNAs can only progress by bringing more statistical rigor to bear in this field and by evaluating changes of individual miRNAs in the context of the overall miRNA network. We have previously performed the first prospective population-based study on circulating miRNAs and demonstrated that loss of endothelial miR-126 is associated with peripheral vascular complications in patients with type 2 diabetes. A proteomics approach was employed to identify potential targets of miR-126. Endothelial cells transfected with the precursor of miR-126 showed reduced secretion of plasminogen-activator inhibitor-1, the key inhibitor of endogenous fibrinolysis. Thus, in subjects with type 2 diabetes differential co-expression patterns of circulating miRNAs occur around endothelial-enriched miR-126. This biomarker association has a potential mechanistic underpinning in the observed regulatory effect of miR-126 on fibrinolysis.
11:30-11:55 Exploring Circulating miRNAs as Blood-Based Diagnostic Biomarkers
Keith W. Jones, Ph.D., Vice President, Biological Sciences, Affymetrix
Effective diagnosis and surveillance of complex multi-factorial disorders such as cancer can be improved by screening of easily accessible biomarkers. Highly stable cell-free circulating nucleic acids (CNA) present as both RNA and DNA species have been discovered in the blood and plasma of humans. microRNAs (miRNAs) representing a class of naturally occurring small non-coding RNAs have emerged as an important set of markers whose concentration in circulation can emulate tissue- and disease-specific expression profiles. In this study we investigate some of the pre-analytic considerations for isolating plasma fractions for the study of miRNA biomarkers. We find that measurement of circulating miRNA levels are frequently confounded by varying levels of cellular miRNAs of different hematopoietic origins and offer an opinion on best practices for isolation and stabilization of circulating miRNAs. We extend this analysis to determining the utility of naturally occurring circulating miRNAs as potential blood-based biomarkers for non-invasive prediction of ulcerative colitis (UC) and define a signature of 31 differentially expressed platelet-derived miRNAs that have a predictive score of 92.8% accuracy, 96.2% specificity and 89.5% sensitivity in distinguishing UC patients from normal individuals. Analysis of these expression maps provide a promising catalog of novel platelet-derived miRNA biomarkers of clinical utility and provide insight into the potential biological function of these candidates in disease pathogenesis.
11:55-12:10 Sponsored Presentation
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12:10-12:35 A Nanopore Sensor for Detection of Circulating microRNAs in Lung Cancer Patients
Li-Qun Gu, Ph.D., Associate Professor, Biological Engineering and Dalton Cardiovascular Research Center, University of Missouri
A nanopore sensor is proposed to selectively detect microRNAs at the single molecular level in plasma samples from lung cancer patients without the need for labeling or amplification. The sensor, which used a programmable oligonucleotide probe to generate a target-specific signature signal, was able to quantify sub-picomolar levels of cancer-associated microRNAs and to discriminate single nucleotide differences between microRNA family members. This approach could be useful for quantitative microRNA detection, biomarker discovery, and the non-invasive early diagnosis of cancer.
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