Nephrology and Transplantation Update Course. Epigenetics and CKD
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1 Nephrology and Transplantation Update Course Epigenetics and CKD Masaomi Nangaku Division of Nephrology and Endocrinology the University of Tokyo Graduate School of Medicine
2 COI disclosure presenter: Masaomi Nangaku I have the following relationships to disclose. Potential Financial Conflicts of Interest (1)Employment: No (2)Stock ownership or options: No (3)Patent royalties/licensing fees: No (4)Honoraria and advisory fees: Kyowa-Hakko-Kirin, Astellas, Chugai, GSK, JT, Tanabe-Mitsubishi (5)Research funding: Kyowa-Hakko-Kirin, Daiichi-Sankyo, Alexion, Astellas, Takeda, JT
3 Mimura, Tanaka, Nangaku. CEPP 2016
4 Epigenetics: the study of persistent changes in gene expression that does not involve mutations of the underlying DNA
5
6 Metabolic memory Epigenetic changes
7 Long-term benefits of intensive glucose control for preventing ESKD: ADVANCE-ON Muh Geot Wong et al. Dia Care 2016
8 5 th September Seminar 2: Basic Science; Diabetes Chairs: Josephine Forbes & Melinda Coughlan Karin Jandeleit-Dahm Masaomi Nangaku Carol Pollock
9 Hypoxic memory Epigenetic changes
10 GFR AKI: No CKD Progression AKI: CKD Progression AKI on CKD: CKD Progression
11 VAP-1 in pericytes enhances neutrophil infiltration into the IR-injured kidney by generating H 2 O 2 Tanaka, Nangaku et al. Kidney Int 2017
12 Basile, Nangaku et al for the ADQI XIII Work Group. JASN 2016
13 Basile, Nangaku et al for the ADQI XIII Work Group. JASN 2016
14 Renal hypoxia in the pathophysiology of the AKI-to-CKD transition Nangaku et al. Nephron 2017
15 Hypoxia Uremia Modified from Mimura, Tanaka, & Nangaku. Semin Nephrol 2013
16 Hypoxia as the final common pathway to ESKD Nangaku. J Am Soc Nephrol 2006
17 Peritubular ischemia contributes more to tubular damage than proteinuria in immune-mediated glomerulonephritis Double immunostaining showed pimonidazole-positive tubules (brown) located close to Kim-1 positive tubules (pink). pimonidazole Muh Geot Wong et al. JASN 2008
18 HIF Defense against hypoxia Nrf2 Defense against oxidative stress Hirakawa, Tanaka, Nangaku. J Diabetes Investig 2017
19 2016 Albert Lasker Basic Medical Research Award Oxygen sensing an essential process for survival William G. Kaelin, Jr. Peter J. Ratcliffe Gregg L. Semenza
20 Hypoxia Uremia Modified from Mimura, Tanaka, & Nangaku. Semin Nephrol 2013
21 D-serine accelerates tubular senescence γ-h2ax staining Chirality Control D-serine SA-βG staining Okada, Nangaku et al. Sci Rep in press
22 Okada, Nangaku et al. Sci Rep in press
23 Indoxyl sulfate suppresses HIF activity reporters HIF targets IS: indoxyl sulfate Tanaka, Nangaku et al. FASEB J 2013
24 Suppression of HIF and its targets by indoxyl sulfate IL-10 VEGF Huang et al. Kidney Int 2016
25 Indoxyl sulfate inhibits binding of HIF to its co-factor, p300 Tanaka, Nangaku et al. FASEB J 2013
26 Indoxyl sulfate induces HIF-inhibiting CITED2 protein CITED2 protein Increased CITED2 mrna stability by IS CITED2 mrna Tanaka, Nangaku et al. FASEB J 2013
27 The efficient displacement of HIF-1α from its complex with CBP/p300 by CITED2 is kinetically driven and proceeds through a transient ternary intermediate Berlow et al. Nature 2017
28 Hypoxia Uremia Modified from Mimura, Tanaka, & Nangaku. Semin Nephrol 2013
29 Measurement of oxygen tension in the kidney utilizing a novel phosphorescence probe Hirakawa, Nangaku et al. Sci Rep 2015
30 Phosphofluorescent probe to detect hypoxia Phosphofluorescence life time (µs) Hirakawa, Nangaku et al. Sci Rep 2015
31 S1 segment has higher oxygen tension compared with S2 segment U: upstream tubules D: downstream tubles Hirakawa & Nangaku. manuscript in submission
32 Epigenetics DNA methylation mirna and lncrna Histone modification Chromosomal conformational change
33 Maternal protein restriction diet induced less weaning weight and hypermethylation of ribosomal DNA in offspring Holland et al. Science 2016
34 Ischemia-reperfusion injury decreased the genomewide methylation level and the CpG methylation level Zhao et al. Gene 2017
35 Aberrant Rasal1 promoter methylation contributes to sustained fibroblast activation and AKI-to-CKD progression Tampe et al. Kidney Int 2017 Rasal1: Ras-Gap like protein-1
36 Epigenetics DNA methylation mirna and lncrna Histone modification Chromosomal conformational change
37 A clear advantage of targeting lncrna rather than epigeneticrelated enzymes or other non-coding RNA such as mirna is the direct sequence specificity of the target site.
38 RNA-seq revealed DARS-AS1 as a hypoxia-induced lncrna Mimura, Nangaku et al. Physiol Rep 2017
39 DARS-AS1 is induced by HIF-1 Mimura, Nangaku et al. Physiol Rep 2017
40 Knockdown of DARS-AS1 aggravates apoptotic cell death Mimura, Nangaku et al. Physiol Rep 2017
41 Epigenetics DNA methylation mirna and lncrna Histone modification Chromosomal conformational change
42 Maternal H3K27me3 controls DNA methylation-independent imprinting Inoue et al. Nature 2017
43 Epigenetic modification of expression of glucose transporter 3 (GLUT3) by hypoxia enhancer mark H3K4me1 normoxia H3K4me1 hypoxia H3K27ac normoxia H3K27ac hypoxia active mark H3K4me3 normoxia H3K4me3 hypoxia HIF1α normoxia HIF1α hypoxia GLUT3-24kb -35kb Mimura, Nangaku et al. Mol Cell Biol 2012
44 Mimura, Nangaku et al. Mol Cell Biol 2012 Histone demethylation by hypoxia Upregulation of KDM3A h hr hrs hrs 8 hrs 12hrs 24hrs hr KDM3A = Jmjd1a demethylase of H3K9me2 OFF ON supprssive H3K9me2
45 Regulation of GLUT3 expression by hypoxia normoxia hypoxia GLUT3 GLUT3 Up-regulation Mimura, Nangaku et al. Mol Cell Biol 2012
46 Cross-enhancement of ANGPTL4 transcription by HIF1 and PPAR β/δ HIF1α PPARβ/δ Inoue, Nangaku et al. Genome Biol 2014
47 Inoue, Nangaku et al. Genome Biol 2014
48 OMX super-resolution microscopy showed changes of the distribution of H3K9Me3 marks in NRK-52e cells by TGF-β1 H3K9Me3: repressive histone mark NUP62: nuclear envelope protein nucleoporin 62 Hewitson et al. Front Pharmacol 2017
49 curcumin improves nephrosclerosis via suppression of histone acetylation acetylated H3K9 Normal salt High salt High salt + curcumin Serum creatinine ChIP using anti-acetyl-h3k9 Muta et al. NDT 2016
50 increased EZH2 in the human fibrotic kidney EZH2 (Enhancer of zeste homolog 2): histone methyltransferase of H3K27me EZH2 mediates kidney fibrosis by downregulating expression of Smad7 and PTEN Zhou et al. JASN 2016
51 Dznep attenuates renal fibrosis in obstructed kidneys Dznep: inhibitor of EZH2 Zhou et al. JASN 2016
52 I/R Vehicle αsma Ab Β-Actin Ab αsma Vehicle Amelioration of AKI-to-CKD transition by Dznep Dznep Dznep Area(%) WB Control * Vehicle群 Vehicle Vehicle 群 * * I/R * p<0.001 * 阻害薬投与群 Dznep Dznep 阻害薬投与群 Mimura, Hirakawa, Nangaku. manuscript in submission
53 H3K27me3 (DAB) Suppression of H3K27me3 by Dznep Vehicle Dznep Nuclear staining of tubular cells showed decreased staining of H3K27me3. Mimura, Hirakawa, Nangaku. manuscript in submission
54 DZnep 2 control 1 control 2 hypoxia 1 hypoxia 2 DZnep 1 DZnep genes Vehicle Control Vehicle I/R injury Dznep I/R injury Group1 Group2 Group3 103 genes149 genes 226 genes Low High RNA-seq of tubules isolated by laser capture microdissection
55 RNA-seq (rpkm) RNA-seq of the kidney Dznep suppressed TIMP2 expression in the kidney 20 Timp Vehicle Ctl Vehicle IR Dznep IR Mimura, Hirakawa, Nangaku. manuscript in submission
56 RNA-seq (rpkm) RNA-seq: in vitro HK2 RPTEC 400 TIMP2 80 TIMP Normoxia Dznep(-) Hypoxia Dznep(-) HK2: human kidney-2 Hypoxia Dznep(+) 0 Normoxia Dznep(-) Hypoxia Dznep(-) Hypoxia Dznep(+) RPTEC: renal proxymal tubular epithelial cell (human primary culture cells) Mimura, Hirakawa, Nangaku. manuscript in submission
57 Dznep suppresses expression of TIMP2 via microrna Ischemia/ reperfusion TIMP2 MMP Collagen Fibrosis Ischemia/ reperfusion TIMP2 MMP Collagen Fibrosis Dznep Novel microrna targets Identified by small RNA-seq Mimura, Hirakawa, Nangaku. manuscript in submission
58 Mimura, Tanaka, & Nangaku. Semin Nephrol 2013
59 ISN President LOC chair Meetings Committee chair
60
61 Stuart Shankland William Couser Richard Johnson Mark Okusa Motoko Yanagita David Harris Kai-Uwe Eckardt Juergen Floege
62 Reiko Inagi (CKD Pathophysiology) Tetsuhiro Tanaka (Nephrology & Endocrinology)
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