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Expression data of Bone Marrow Derived Macrophages (BMDM) from WT and Nos2-/- mice proinflammatory activated in vitro

ABSTRACT: Proinlfammatory stimulation of murine macrophages elicits profound metabolic changes. iNOS-derived Nitric oxide is both necessary and sufficient for the repression of OXPHOS during M1 polarization. Macrophages derived from mice lacking NOS2 display intact metabolic pathway and fail to rewire carbon fluxes. Since inflammation results in a very controlled genetic programme where specific sets of genes are up-regulated we aim to understand how this is affected in absence of this important endogenous gas responsible of the metabolic phenotype. We used microarrays to detail the global programme of gene expression underlying proinflammatory stimulation and identified distinct classes of differentially regulated genes during this process in absence of NOS2

ORGANISM(S): Mus musculus

PROVIDER: GSE115120 | GEO | 2019/12/10

REPOSITORIES: GEO

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Protein Modifications Driven by Nitric Oxide and Nitroxyl during Macrophage Proinflammatory Activation

Project description:Proinlfammatory stimulation of murine macrophages elicits profound metabolic changes. iNOS-derived Nitric oxide (NO) is both necessary and sufficient for the repression of OXPHOS and major mitochondrial rewiring of carbon fluxes during M1 polarization. Since inflammation results in broad proteomics changes with oxidative modifications of specific targets, we aim to understand how this is managed in absence of this important endogenous gas and its derived reductive nitrogen species like nitroxyl (HNO). We used proteomics to detail the mitochondrial proteome expression and status of cysteine modifications underlying proinflammatory stimulation and identified distinct targets of reversible and irreversible cysteine oxidations during this process in presence and absence of NOS2, RNS donors, and in exacerbating conditions of oxidative stress.

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Project description:Inducible nitric oxide synthase (iNOS) plays a crucial role in controlling growth of mycobacteria, presumed to be via nitric oxide (NO) mediated killing. However, NOS enzymes can also signal through NO-independent pathways, and production of NO by NOS requires the cofactor tetrahydrobiopterin (BH4). We compared Nos2-/- mice to mice with macrophage BH4 deficiency (Gch1fl/flTie2cre), due to a leukocyte-specific deletion of Gch1, to uncover the specific contribution of NO-independent NOS functions to anti-mycobacterial immunity. We used microarrays to detail the global programme of gene expression in uninfected and BCG infected macrophages that were either deficient in iNOS (Nos2-/- vs C57bl6/J) or BH4/Gch1 (GCHfl/flTie2cre vs GCHfl/fl)

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Project description:Tristetraprolin (TTP), encoded by the Zfp36 gene, is a zinc-finger protein that regulates RNA stability primarily through association with 3’ untranslated regions (3’UTRs) of target mRNAs. While TTP is expressed abundantly in the intestines, its function in intestinal epithelial cells (IECs) is unknown. Here we used a cre-lox system to remove Zfp36 in the mouse epithelium to uncover a role for TTP in IECs and to identify target genes in these cells. While TTP was largely dispensable for establishment and maintenance of the colonic epithelium, we found an expansion of the proliferative zone and an increase in goblet cell numbers in the colon crypts of Zfp36ΔIEC mice. Furthermore, through RNA-sequencing of transcripts isolated from the colons of Zfp36fl/fl and Zfp36ΔIEC mice, we found that expression of inducible nitric oxide synthase (iNos or Nos2) was elevated in TTP-knockout IECs. We demonstrate that TTP interacts with AU-rich elements in the Nos2 3’UTR and suppresses Nos2 expression. In comparison to control Zfp36fl/fl mice, Zfp36ΔIEC mice were less susceptible to dextran sodium sulfate (DSS)-induced acute colitis. Together, these results demonstrate that TTP targets Nos2 expression in IECs and aggravates acute colitis.

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