Expression data from wild type or FIZZ2 knockout murine lungs
ABSTRACT: To study the possible fibrotic role of FIZZ2, bleomycin was used to induce pulmonary fibrosis in wild type and FIZZZ2 knockout mice, lungs were then harvested and processed for RNA isolation. We used microarrays to detail the global gene expression regulated by FIZZ2 during fibrotic process. Overall design: Bleomycin/saline treated wild type or FIZZ2 knockout lungs at day 21 post injection were harvested for RNA isolation and hybridization on Affymetrix microarrays
Project description:To study the possible fibrotic role of FIZZ2, bleomycin was used to induce pulmonary fibrosis in wild type and FIZZZ2 knockout mice, lungs were then harvested and processed for RNA isolation. We used microarrays to detail the global gene expression regulated by FIZZ2 during fibrotic process. Bleomycin/saline treated wild type or FIZZ2 knockout lungs at day 21 post injection were harvested for RNA isolation and hybridization on Affymetrix microarrays
Project description:Lung fibroblasts play a pivotal role in pulmonary fibrosis, a devastating lung diseases, by producing extracellular matrix. MicroRNAs (miRNAs) suppress a lot of genes posttranscriptionally, but the dynamics and the role of miRNAs in activated lung fibroblasts in fibrotic lung has been poorly understood. To elucidate these problems, we performed global miRNA expression profiling of lung fibroblasts of bleomycin- and silica-induced fibrotic lungs Overall design: Global miRNA expression profile of lung fibroblasts isolated from untreated, bleomycin-treated and silica-treated mice on days 7, 14 and 28 post-treatment was generated using Ion Total RNA-seq Kit v2 and Ion PGM sequencer.
Project description:We have employed whole genome microarray expression profiling as a discovery platform to identify genes with the potential to distinguish Mmp19 regulation of fibroblast phenotype changes in mouse lungs. Pulmonary fibrosis was induced by bleomycin at 0.08 u in 50ul of saline. At 21st day the mice were sacrificed and mouse lung fibroblasts were isolated and cultured in FBM plus additives following Lonza's portocol. RNA was extracted with miRNA mini kit from Qiagen. Gene expression microarray was performed with Agilent. A 834-gene consensus signature was identified that distinguished between Mmp19 knockout mice from wildtype. Some gene expression in the same RNA samples were validtaed by real-time PCR. The established bleomycin induced fibrosis was used in this experiment. At day 21 the fibrosis would be the situation of stable fibrosis. We administrated 0.08u of bleomycin intratracheally into wildtype and Mmp19 knockout mice, sacrificed the mice at 21st day and isolated the lung fibroblasts and culturing. Five independent experiments were performed and 3 for gene expression experiment.
Project description:Idiopathic pulmonary fibrosis is a devastating aging-associated disease of unknown etiology. Despite that aging is a major risk factor, the mechanisms linking aging with this disease are uncertain, and experimental models to explore them in lung fibrosis are scanty. We examined the fibrotic response to bleomycin-induced lung injury in Zmpste24-deficient mice, which exhibit nuclear lamina defects developing accelerated aging. We found that young WT and Zmpste24(-/-) mice developed a similar fibrotic response to bleomycin. Unexpectedly, while old WT mice developed severe lung fibrosis, accelerated aged Zmpste24-/- mice were protected showing scant lung damage. To investigate possible mechanisms associated with this resistance to fibrosis, we compared the transcriptome signature of the lungs and found that Zmpste24(-/-) mice showed downregulation of several core and associated matrisome genes compared with WT mice. Interestingly, some microRNAs that target extracellular matrix molecules such as miR23a, miR27a, miR29a, miR29b-1,miR145a, and miR491 were dysregulated resulting in downregulation of profibrotic pathways such as TGF-β/SMAD3/NF-κB and Wnt3a/β-catenin signaling axis. These results indicate that the absence of Zmpste24 in aging mice results in impaired lung fibrotic response after injury, which is likely associated to the dysregulation of fibrosis-related miRNAs. Aging is a driving force of pulmonary fibrosis. Mice laking Zmpste24 have an accelerated aging phenotype. During the development of the disease there are several extracellular matrix genes, principally collagens upregulated. We use microarray to unveil changes associated with the pulmonary response in aged mice Overall design: Lung samples 21 days after bleomycin injury were selected for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain changes associated with the fibrotic response in young WT and Zmpste24 KO mice and old WT and Zmpste24 KO ones 21 days after administratiuon of bleomycin. We used three biological replicates for each condition.
Project description:Idiopathic pulmonary fibrosis (IPF) is a complex disease involving various cell types. Macrophages are essential in maintenance of physiological homeostasis, wound repair and fibrosis in the lung. Macrophages play a crucial role in repair and remodeling by altering their phenotype and secretory pattern in response to injury. The secretome of induced pluripotent stem cells (iPSC-cm) attenuates injury and fibrosis in bleomycin injured rat lungs. In the current study, we evaluate the effect of iPSC-cm on interstitial macrophage gene expression and phenotype in bleomycin injured rat lungs in vivo. iPSC-cm was intratracheally instilled 7 days after bleomycin induced lung injury and assessed 7 days later and single cell isolation was performed. Macrophages were FACS sorted and microarray analysis was performed. We characterized changes in the rat lung interstitial macrophages using transcriptional profiling.
Project description:Idiopathic pulmonary fibrosis is a chronic devastating disease of unknown etiology. No therapy is currently available. A growing body of evidence supports the role of TGFβ1 as the major player in the pathogenesis of the disease. This study designed novel human- and mouse-specific siRNAs and siRNA/DNA chimeras targeting both human and mouse common sequences and evaluated their inhibitory activity in pulmonary fibrosis induced by bleomycin and lung-specific transgenic expression of human TGFβ1. Selective novel sequences of siRNA and siRNA/DNA chimeras efficiently inhibited pulmonary fibrosis, indicating their applicability as tools for treating fibrotic disease in humans. Total RNA was extracted from lung tissue from mice with bleomycin (BLM)-induced lung fibrosis treated with mouse TGFβ1 siRNAs or vehicle on different days after BLM infusion.
Project description:Wildtype mice were given saline or bleomycin by oropharyngeal instillation. After 14 days, during the fibrotic phase of the response, lungs were dissected and total RNA was extracted and used for gene expression profiling. The aim was to identify those genes regulated during the development of fibrosis in this animal model of bleomycin-induced lung fibrosis. Wildtype male C57Bl/6J mice (8-10 weeks old) were used in the study, with three mice per group. Bleomycin (1mg/kg body weight in 50μl of saline) or saline was administered by oropharyngeal installation as described previously by Lakatos et al, Exp Cell Res, 2006, under light halothane-induced anaesthesia. After 14 days, lungs were removed, blotted dry and the trachea and major airways were excised before the separated lobes were snap frozen in liquid nitrogen. Lungs were then pulverized under liquid nitrogen, and the resulting lung powder was used for total RNA extraction using Trizol. Following DNase-treatment, clean-up, cDNA synthesis and cRNA synthesis, samples were hybridized to Affymetrix MOE430A genechips using standard protocols. The data were analyzed using RMA with quantiles normalization.
Project description:Pulmonary hypertension is a frequent consequence of left heart disease and congestive heart failure (CHF) and causes extensive lung vascular remodelling which leads to right ventricular failure. Functional genomics underlying this structural remodelling are unknown but present potential targets for novel therapeutic strategies. We used microarrays to detail the gene expression underlying vascular remodeling in the pathogenesis of pulmonary hypertension and identified distinct classes of up-regulated genes during this process. Overall design: Control rat lung samples were compared to samples of aortic banding rat lungs which exhibit pulmonary hypertension