title: An integrated nano-scale approach to profile miRNAs in limited clinical samples.
abstract: Profiling miRNA expression in cells that directly contribute to human disease pathogenesis is likely to aid the discovery of novel drug targets and biomarkers. However, tissue heterogeneity and the limited amount of human diseased tissue available for research purposes present fundamental difficulties that often constrain the scope and potential of such studies. We established a flow cytometry-based method for isolating pure populations of pathogenic T cells from bronchial biopsy samples of asthma patients, and optimized a high-throughput nano-scale qRT-PCR method capable of accurately measuring 96 miRNAs in as little as 100 cells. Comparison of circulating and airway T cells from healthy and asthmatic subjects revealed asthma-associated and tissue-specific miRNA expression patterns. These results establish the feasibility and utility of investigating miRNA expression in small populations of cells involved in asthma pathogenesis, and set a precedent for application of our nano-scale approach in other human diseases. The microarray data from this study (Figure 7) has been submitted to the NCBI Gene Expression Omnibus (GEO; http://ncbi.nlm.nih.gov/geo) under accession no. GSE31030.
authors.affiliation: Sandler Asthma Basic Research Center, University of California San Francisco, San Francisco, CA, USA ; Division of Infection, Inflammation and Immunity, University of outhampton, School of Medicine, Southampton NIHR Respiratory Biomedical Research Unit, Sir Henry Wellcome Laboratories, Southampton General Hospital, Southampton, UK ; Pulmonary and Critical Care Division, Department of Medicine, University of California San Francisco.
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