MicroRNA Expression Profile in Exhaled Breath Condensate: Investigating the Pathophysiology of Idiopathic Pulmonary Fibrosis
Priya Agarwala, Nadia Pletukhina, Lora J. Kasselman, Heather A. Renna, Daniel S. Glass, Steve H. Salzman, Allison B. Reiss. Medicine, NYU Winthrop Hospital, Mineola, New York, United States
Purpose of Study Idiopathic pulmonary fibrosis (IPF) is an irreversible fibrotic lung disease which ultimately results in respiratory failure. It is incurable, the etiology is unknown and therapeutic options are limited. Exhaled breath condensate (EBC) composed of droplets of airway epithelial lining fluid and condensed water vapor contains variety of biomolecules, including micro(mi)RNAs, with potential for non-invasive monitoring of pathological processes in the lungs. EBC miRNA may be useful in advancing IPF disease diagnosis, staging, understanding of pathophysiology and even as possible therapeutic targets. This study investigates the microRNA composition of EBC from patients with IPF compared to healthy controls in order to detect abnormal miRNA expression in IPF that may be leveraged to develop new treatments.
Methods Used EBC was collected using a portable non-invasive handheld device for the study of deep-lung volatile oranic compounds. miRNA extraction from EBC was performed with the Qiagen miRNA isolation kit. The extracted miRNA was digested with DNase I to remove genomic DNA contamination and quantified by spectrophotometry. Isolated miRNA was converted to cDNA with reverse transcriptase and the resulting cDNA was plated in a miScript miRNA PCR Human Fibrosis array.
Summary of Results Data from the miScrpt miRNA PCR array show that there is a clear difference in microRNA expression between patients with and without IPF and these specific microRNAs have been identified. In particular, several fibrosis-related microRNAs are overexpressed in IPF patients including: miR-10a, miR-142, miR-204, and miR-20a.
Conclusions The miRNA profile of EBC from IPF patients differs from that of controls matched by age and sex. This information will be applied in culture models of alveolar epithelial cells to study effects on fibrosis of overexpression and silencing of these outlier miRNAs. Downstream effects of these miRNAs on cellular function will then be evaluated in order to reveal new therapeutic targets.
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