Idiopathic Pulmonary Fibrosis (IPF) is a chronic progressive lung disease. This application will examine the hypothesis that epigenetic dysregulation underpins pulmonary fibrosis. We will examine miRs-19b and -20a, two members of the miR-17~92, which target DNA methyltransferase-1, DNMT1. We found that miRs-19b and -20a expression are reduced in the lung tissue of patients with IPF, accompanied by increased expression of DNMT1. Thus, the grant application investigates the role of DNMT1 in IPF and the relationship of miRs-19b and - 20a in this regulation. Moreover, we believe that the cluster itself is also regulated by DNA methylation of CpG islands in its promoter, forming a negative feedback loop between DNMT1 and miRs-19b and - 20a expression. In this proposal, we will use in vitro and in vivo studies to assess this hypothesis. To address our hypothesis in vivo, we have generated cell-specific miR-17~92 and DNMT1 knockdowns in transgenic murine models. We will also examine whether miRs-19b and -20a regulates components involved in epigenetic regulation. We anticipate that understanding the relationship between miRs-19b and -20a and DNMT1 expression in lung fibrosis will identify novel therapeutic strategies and underlying mechanisms of fibrosis. We predict that our findings will lead to the development of human clinical trials. To accomplish these goals, we propose the following two specific aims:
Specific Aim 1) Understand the role of DNMT1 in expression and methylation of the miR-17~92 cluster in vitro and the reciprocal role of miRs-19b and -20a on DNMT1 expression.
Specific Aim 2) Determine the in vivo role of DNMT1 in pulmonary fibrosis.
There are significant gaps in understanding the pathogenesis of Idiopathic Pulmonary Fibrosis (IPF) at the cellular and genetic levels. This disease is associated with high mortality due to the lack of therapies. We have found changes in regulatory proteins that are critical for gene expression and repair of lung fibrosis. Interestingly, current FDA approved treatments for hematological malignancies target some of these proteins. Thus, in this application, we propose in vitro and in vivo pre-clinical testing to determine the efficacyof these potential treatments. We anticipate that our findings will lead to the development of therapies to individuals with IPF.
|Schilter, Heidi; Cantemir-Stone, Carmen Z; Leksa, Vladimir et al. (2015) The mannose-6-phosphate analogue, PXS64, inhibits fibrosis via TGF-Î²1 pathway in human lung fibroblasts. Immunol Lett 165:90-101|
|Forget, Mary A; Voorhees, Jeffrey L; Cole, Sara L et al. (2014) Macrophage colony-stimulating factor augments Tie2-expressing monocyte differentiation, angiogenic function, and recruitment in a mouse model of breast cancer. PLoS One 9:e98623|
|Moldovan, Leni; Batte, Kara E; Trgovcich, Joanne et al. (2014) Methodological challenges in utilizing miRNAs as circulating biomarkers. J Cell Mol Med 18:371-90|
|Blackwell, Timothy S; Tager, Andrew M; Borok, Zea et al. (2014) Future directions in idiopathic pulmonary fibrosis research. An NHLBI workshop report. Am J Respir Crit Care Med 189:214-22|
|Ismail, Noura; Wang, Yijie; Dakhlallah, Duaa et al. (2013) Macrophage microvesicles induce macrophage differentiation and miR-223 transfer. Blood 121:984-95|
|Dakhlallah, Duaa; Batte, Kara; Wang, Yijie et al. (2013) Epigenetic regulation of miR-17~92 contributes to the pathogenesis of pulmonary fibrosis. Am J Respir Crit Care Med 187:397-405|
|Moldovan, Leni; Batte, Kara; Wang, Yijie et al. (2013) Analyzing the circulating microRNAs in exosomes/extracellular vesicles from serum or plasma by qRT-PCR. Methods Mol Biol 1024:129-45|
|Manolio, Teri A; Chisholm, Rex L; Ozenberger, Brad et al. (2013) Implementing genomic medicine in the clinic: the future is here. Genet Med 15:258-67|
|Wenzke, Kevin E; Cantemir-Stone, Carmen; Zhang, Jie et al. (2012) Identifying common genes and networks in multi-organ fibrosis. AMIA Jt Summits Transl Sci Proc 2012:106-15|
|Payne, Philip R O; Marsh, Clay B (2012) Towards a "4I" approach to personalized healthcare. Clin Transl Med 1:14|