title: Renal ischemia-reperfusion injury upregulates histone-modifying enzyme systems and alters histone expression at proinflammatory/profibrotic genes.
abstract: Ischemic renal injury can produce chronic renal inflammation and fibrosis. This study tested whether ischemia-reperfusion (I/R) activates histone-modifying enzyme systems and alters histone expression at selected proinflammatory/profibrotic genes. CD-1 mice were subjected to 30 min of unilateral I/R. Contralateral kidneys served as controls. At 1, 3, or 7 days of reflow, bilateral nephrectomy was performed. Renal cortices were probed for monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-beta1 (TGF-beta1), and collagen III mRNAs and cytokine levels. RNA polymerase II (Pol II) binding, which initiates transcription, was quantified at exon 1 of the MCP-1, TGF-beta1, collagen III genes (chromatin immunoprecipitation assay). Two representative gene-activating histone modifications [histone 3 lysine 4 (H3K4) trimethylation (m3) (H3K4m3); histone 2 variant H2A.Z] were sought. Degrees of binding of two relevant histone-modifying enzymes (Set1, BRG1) to target genes were assessed. Renal cortical Set1, BRG1, and H2A.Z mRNAs were measured. Finally, the potential utility of urinary mRNA concentrations as noninvasive markers of these in vivo processes was tested. I/R caused progressive increases in Pol II binding to MCP-1, TGF-beta1, and collagen III genes. Parallel increases in cognate mRNAs also were expressed. Progressive increases in renal cortical Set1, BRG1, H2A.Z mRNAs, and increased Set1/BRG1 binding to target genes occurred. These changes corresponded with: 1) progressive elevations of H3K4m3 and H2A.Z at each test gene; 2) increases in renal cortical TGF-beta1/MCP-1 cytokines; and 3) renal collagen deposition (assessed by histomorphology). Postischemic increases in urinary TGF-beta1, MCP-1, Set1, and BRG1 mRNAs were also observed. We conclude that: 1) I/R upregulates histone-modifying enzyme systems, 2) histone modifications at proinflammatory/profibrotic genes can result, and 3) urinary mRNA assessments may have utility for noninvasive monitoring of these in vivo events.
authors.affiliation: Department of Medicine, University of Washington, and the Clinical Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, WA 98109, USA. dzager@fhcrc.org
authors.forename: Richard A
authors.initials: RA
authors.lastname: Zager
authors.forename: Ali C M
authors.initials: AC
authors.lastname: Johnson
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000652:urine; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics; D024061:Collagen Type III / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics; D005355:Fibrosis; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics; D006657:Histones / Q000378:metabolism*; D007672:Kidney Cortex / Q000201:enzymology* / Q000473:pathology; D007674:Kidney Diseases / Q000235:genetics / Q000473:pathology / Q000503:physiopathology*; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000201:enzymology; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009393:Nephritis / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D009687:Nuclear Proteins / Q000235:genetics; D012319:RNA Polymerase II / Q000378:metabolism; D012333:RNA, Messenger / Q000652:urine; D015427:Reperfusion Injury / Q000235:genetics / Q000473:pathology / Q000503:physiopathology*; D014157:Transcription Factors / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000235:genetics; D015854:Up-Regulation / Q000502:physiology
chemical_list: D015415:Biomarkers; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D024061:Collagen Type III; D006657:Histones; D009687:Nuclear Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D053773:Transforming Growth Factor beta1; D011495:Histone-Lysine N-Methyltransferase; D012319:RNA Polymerase II; C513194:Smarca4 protein, mouse; D004265:DNA Helicases