Project description:Purpose: Compare the transciptome between lung myofibroblasts that contituatively express Fra-2 to control lung myofibroblasts to determine genes that may contribute to regulation of secondary alveolar septation Methods: mRNa sequences from lung myofibroblasts isolated from mutant mice that contituatively express Fra-2 in α-SMA expressing cells (sma-Fra2) were compared to control mice. n=4 in each group Results: 43,432 transcripts wre analyzed and 361 genes were differentially expressed in the smaFra2 mice compared to wild type control. Functional analysis was performed using the Database for Annotation, Visualization and Integrated Discovery. Changes in genes from lung myofibroblasts in smaFra2 mice: genes involved with extracellular matrix and cell adhesion were upregulated; Ectonucleotide pyrophosphatase/phosphodiesterase family member 2 (Enpp2) and LPA receptor1 (LPA1) were downregulated. Coclusion: The secondary alveolar septation defect in smaFra2 mice could be explaned by Fra-2 mediated changes in extracellular matrix and cell adhesion in developing lung myofibroblasts.
Project description:The Mg-delta strain of fibrillin-1 deficient mice display an early defect in alveolar septation. This study attempts to identify expression patterns in the mutant mouse lung compared to their wild-type littermates that suggest pathways that are critical for normal septation. This experiment focuses on postnatal days 1 and 5. Other time points will be added at a later date.
Project description:The Mg-delta strain of fibrillin-1 deficient mice display an early defect in alveolar septation. This study attempts to identify expression patterns in the mutant mouse lung compared to their wild-type littermates that suggest pathways that are critical for normal septation. This experiment focuses on postnatal days 1 and 5. Other time points will be added at a later date. Keywords: other
Project description:The septation initiation network (SIN), composed of a conserved SepH (Cdc7p) kinase cascade, plays an essential role in fungal cytokinesis/septation and conidiation for asexual reproduction, while the mitogen-activated protein kinase (MAPK) pathway depends on successive signaling cascade phosphorylation to sense and respond to stress and environmental factors. In this study, a SepH suppressor–PomA in the filamentous fungus A. nidulans was identified as a negative regulator of septation and conidiation such that the pomA mutant was able to cure defects of sepH in septation and conidiation and overexpression of pomA remarkably suppressed septation. Under the normal cultural condition, SepH positively regulates the phosphorylation of MAPK-HogA, while PomA reversely affects this process and the phosphorylated HogA in MAPK could be a downstream product of the activated SIN. However, under the osmostress condition, the induced phosphorylated HogA is capable of bypassing the requirement of SepH to fulfill septation and conidiation. Findings demonstrate that crosstalk exists between the SIN and MAPK pathways.
Project description:Regulation of lung responses to air breathing and continued alveolar development shape ex-utero and adult respiratory health. Applying single-cell transposome hypersensitive site sequencing (scTHS-seq) to human and mouse lung, we assembled a regulatory atlas of postnatal lung development and analyzed over 80,000 single-cells. From day of life 1, we characterized cell types and cell-cell interactions driving alveoli generation. We elucidated key transcription factor-gene accessibility modules (TF-GAM) guiding alveolar epithelial cell differentiation and epithelial-mesenchymal interactions involved in alveolar septation and mechanotransduction. Incorporating GWAS causal variants, we identified lung pathogenic risk units enriched in specific cell types. Our regulatory map and analysis model provide an effective resource to investigate age-dependent and species-specific control of critical developmental processes. Furthermore, this resource complements single-cell atlas efforts to advance our understanding of lung health and disease across the human lifespan.
Project description:In the paper "Fra-1 regulates its target genes via binding to remote enhancers without exerting major control on chromatin architecture in triple negative breast cancers" by Bejjani et al., we identified Fra-1 and/or Fra-2 target genes in MDA-MB-231 cells. si RNA against Fra-1 and against Fra-2 were transfected in MDA-MB-231 cells either independenlty or simultaneously to identify genes regulated specifically by Fra-1 or Fra-2 and genes regulated redundantly or complementarily by Fra-1 and Fra-2 total RNA were purified and biotinylated sense-strand cDNA were produced. cDNA targets were used to probe Affymetrix GeneChip Human Gene 2.0 ST arrays
Project description:Lung alveolarization is a complex process that involves interactions between several cell types and leads to considerable increase in gas-exchange surface area. The step designated secondary septation includes elastogenesis from interstitial fibroblasts. We used microarrays to detail the global programme of fibroblast gene expression that underly secondary septation, and identified numerous genes that undergo signficant changes in expression Experiment Overall Design: Rat lung fibroblats were extemporaneously isolated at successive stages of postnatal development that flank the septation period for RNA extraction and hybridization on Affymetrix microarrays. Three biological replicates from three different litters were prepared for each time point.
Project description:In the article "Fra-1 regulates its target genes via binding to remote enhancers without exerting major control on chromatin architecture in triple negative breast cancers" by Bejjani et al., we mapped epigenetic marks (H3K4me1, H3K4me3, H3K27ac), p300/CBP, PolII and CTCF to characterize the binding sites of Fra-1 and Fra-2 on MDA-MB-231 genome. Data for Fra-1 and Fra-2 ChIP-seq are available on GEO database, accession number GSE132098 (Tolza et al., 2019, MCR 17, 1999-2014)
Project description:It has been shown that dexamethasone (Dex) impairs the normal lung septation that occurs in the early postnatal period. Treatment with retinoic acid (ATRA) abrogates the effects of Dex. To understand the molecular basis for the Dex indiced inhibition of the formation of the alveoli and the ability of ATRA to prevent the inhibition of septation, gene expression was analyzed in 4-day old mice treated with diluent (control), Dex-treated and ATRA+Dex-treated.
