SESSION TITLE: Lung Cancer II
SESSION TYPE: Slide Presentations
PRESENTED ON: Sunday, March 23, 2014 at 12:15 PM - 01:15 PM
PURPOSE: Biomarkers are of importance for understanding the biology of disease and in diagnosis and treatment planning. MicroRNAs are short non-coding ribonucleotide chains which modify the expression of multiple genes and pathways. MicroRNAs in BAL and bronchial washings may prove useful as prognostic/ predictive and monitoring markers. However, microRNA measurements in these samples have yet to be established. We present data from a pilot study in which microRNAs implicated in lung cancer were measured.
METHODS: Lavage was performed as clinically indicated, filtered, and analysed according to standard clinical practice. The remaining fluid was frozen in 0.5-1.0ml aliquots at -20 °C. MicroRNAs were measured using qPCR. After precipitation and removal of the cellular fraction, two 200mcl aliquots per patient were analysed using six qPCR assays, each in duplicates. Spike-in miRNAs (C. elegans) were introduced before RNA extraction for standardized assessment of RNA recovery. In addition, RNA U6B was measured as a putative internal control RNA.
RESULTS: Lavage samples from four patients with sarcoidosis, lung adenocarcinoma, pulmonary fibrosis and chronic LTX rejection were analysed for miR-142 3p, miR-205, miR-486. Spike-in C. elegans miRNAs -39 and -54 were detected at a constant level in all samples. The RNA U6B was only weakly expressed. The target miRNAs were measurable in all samples. Technical replicates of the qPCR assays gave consistent results in all samples. Analyses of two aliquots from the same patient resulted in only minimal differences in measured miRNA levels. Measurements of miR-142-3p in the sarcoidosis aliquots showed slightly more deviation than in the remaining paired samples.
CONCLUSIONS: MicroRNAs could be identified reliably and reproducibly in all lavage samples. MiR-142 3p, miR-205 and miR-486 were selected based on their role in serum studies of NSCLC.
CLINICAL IMPLICATIONS: Further investigations of the significance of local microRNAs at diagnosis and during the course of treatment are ongoing.
DISCLOSURE: S. Kaduthanam: Grant monies (from sources other than industry): grant from federal ministry Ruprecht Kuner: Grant monies (from sources other than industry): grant from federal ministry The following authors have nothing to disclose: Amanda Tufman, Fernando Gamarra, Rosi Kiefl, Rudolf Huber
research on potential biomarkers and pathogenesis of lung diseases