Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture

The development of whole-transcriptome microarrays and next-generation sequencing has revolutionized our understanding of the complexity of cellular gene ...

University of Cambridge

&#x8840;&#x7EA2;&#x7D20;&#x52A0;&#x6C27;&#x9176;-1&#xFF0C;&#x6709;&#x8FB1;&#x4EBA;&#x683C;&#x5BF9;&#x4E00;&#x6C27;&#x5316;&#x78B3;&#x3001; &#x94C1;&#x548C;&#x80C6;&#x7EFF;&#x7D20;&#xFF0C;&#x8840;&#x7EA2;&#x7D20;&#x9176;&#x5DF2;&#x88AB;&#x516C;&#x8BA4;&#x4E3A;&#x5BF9;&#x538B;&#x529B;&#x7684;&#x6761;&#x4EF6;&#xFF0C;&#x4E0D;&#x53EA;&#x4E0E;&#x76F8;&#x5173;&#x7684;&#x8840;&#x7EA2;&#x7D20;&#x91CA;&#x653E;&#x7EC6;&#x80DE;&#x9632;&#x5FA1;&#x4E2D;&#x53D1;&#x6325;&#x7740;&#x5173;&#x952E;&#x4F5C;&#x7528;&#x3002;&#x4F55; 1 &#x4FDD;&#x62A4;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x51CB;&#x4EA1;&#x3001; &#x53C2;&#x4E0E;&#x8840;&#x7BA1;&#x653E;&#x677E;&#x8C03;&#x8282;&#x7740;&#x8840;&#x7BA1;&#x5F20;&#x529B;&#x3001; &#x6291;&#x5236;&#x8840;&#x7BA1;&#x58C1;&#x708E;&#x75C7;&#x53CD;&#x5E94;&#x548C;&#x53C2;&#x4E0E;&#x8840;&#x7BA1;&#x5F62;&#x6210;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x548C;&#x8840;&#x7BA1;&#x53D1;&#x751F;&#x3002;&#x540E;&#x8005;&#x7684;&#x804C;&#x80FD;&#x5C06;&#x4F55;-1 &#x7684;&#x94FE;&#x63A5;&#xFF0C;&#x4E0D;&#x4EC5;&#x5BF9;&#x5FC3;&#x8840;&#x7BA1;&#x7F3A;&#x8840;&#x5230;&#x8BB8;&#x591A;&#x5176;&#x4ED6;&#x6761;&#x4EF6;&#xFF0C;&#x5982;&#x53D1;&#x5C55;&#x3001; &#x4F24;&#x53E3;&#x6108;&#x5408;&#xFF0C;&#x6216;&#x764C;&#x75C7;&#xFF0C;&#x4F9D;&#x8D56;&#x4E8E;&#x65B0;&#x751F;&#x8840;&#x7BA1;&#x3002;&#x8FD9;&#x6B21;&#x5168;&#x9762;&#x5BA1;&#x67E5;&#x7684;&#x76EE;&#x7684;&#x662F;&#x89E3;&#x51B3;&#x4F55; 1 &#x8C03;&#x63A7;&#x548C;&#x529F;&#x80FD;&#x7684;&#x5FC3;&#x8840;&#x7BA1;&#x751F;&#x7406;&#x5B66;&#x4E0E;&#x75C5;&#x7406;&#x673A;&#x5236;&#x5E76;&#x8BC1;&#x660E;&#x67D0;&#x4E9B;&#x53EF;&#x80FD;&#x7684;&#x5E94;&#x7528;&#xFF0C;&#x5230;&#x76EE;&#x524D;&#x4E3A;&#x6B62;&#x4EA7;&#x751F;&#x7684;&#x5927;&#x91CF;&#x77E5;&#x8BC6;&#x3002;&#x6700;&#x8FD1;&#x6570;&#x636E;&#x63D0;&#x4F9B;&#x4E86;&#x6709;&#x529B;&#x7684;&#x8BC1;&#x636E;&#xFF0C;&#x4F55; 1 &#x53C2;&#x4E0E;&#x7528;&#x4E8E;&#x5FC3;&#x8840;&#x7BA1;&#x75BE;&#x75C5;&#x7684;&#x836F;&#x7269;&#x7684;&#x7597;&#x6548;&#x3002;&#x5C0F;&#x8BF4;&#x7814;&#x7A76;&#x6253;&#x5F00;&#x5E94;&#x7528;&#x4F55;-1 &#x7684;&#x57FA;&#x56E0;&#x548C;&#x7EC6;&#x80DE;&#x6CBB;&#x7597;&#x7684;&#x53EF;&#x80FD;&#x6027;&#x3002;&#x56E0;&#x6B64;&#xFF0C;&#x5728;&#x4ECA;&#x540E;&#x51E0;&#x5E74;&#x7684;&#x7814;&#x7A76;&#x5E94;&#x6709;&#x52A9;&#x4E8E;&#x6F84;&#x6E05;&#x4F55; 1 &#x836F;&#x7269;&#x7684;&#x7597;&#x6548;&#x548C;&#x57FA;&#x4E8E;&#x4F55;-1 &#x7684;&#x7EC6;&#x80DE;&#x548C;&#x57FA;&#x56E0;&#x6CBB;&#x7597;&#x7684;&#x4E34;&#x5E8A;&#x53EF;&#x884C;&#x6027;&#x7684;&#x771F;&#x6B63;&#x4F5C;&#x7528;&#x4E3A;&#x6B64;&#x521B;&#x5EFA;&#x6709;&#x6548;&#x7684;&#x6CBB;&#x7597;&#x9014;&#x5F84;&#x7CBE;&#x5236;&#x6297;&#x6C27;&#x5316;&#x7CFB;&#x7EDF;&#x3002;

&#x8840;&#x7EA2;&#x7D20;&#x52A0;&#x6C27;&#x9176;-1 &#x548C;&#x8840;&#x7BA1;&#x5E8A;&#xFF1A; &#x4ECE;&#x5206;&#x5B50;&#x673A;&#x5236;&#x5230;&#x6CBB;&#x7597;&#x7684;&#x673A;&#x4F1A;&#x3002;

Heme oxygenase-1, an enzyme degrading heme to carbon monoxide, iron, and biliverdin, has been recognized as playing a crucial role in cellular defense against stressful conditions, not only related to heme release. HO-1 protects endothelial cells from apoptosis, is involved in blood-vessel relaxation regulating vascular tone, attenuates inflammatory response in the vessel wall, and participates in blood-vessel formation by means of angiogenesis and vasculogenesis. The latter functions link HO-1 not only to cardiovascular ischemia but also to many other conditions that, like development, wound healing, or cancer, are dependent on neovascularization. The aim of this comprehensive review is to address the mechanisms of HO-1 regulation and function in cardiovascular physiology and pathology and to demonstrate some possible applications of the vast knowledge generated so far. Recent data provide powerful evidence for the involvement of HO-1 in the therapeutic effect of drugs used in cardiovascular diseases. Novel studies open the possibilities of application of HO-1 for gene and cell therapy. Therefore, research in forthcoming years should help to elucidate both the real role of HO-1 in the effect of drugs and the clinical feasibility of HO-1-based cell and gene therapy, creating the effective therapeutic avenues for this refined antioxidant system.

Heme oxygenase-1 and the vascular bed: from molecular mechanisms to therapeutic opportunities.

H&egrave;me oxyg&eacute;nase-1, une enzyme d&eacute;gradant l&#39;h&egrave;me en monoxyde de carbone, le fer et la biliverdine, a &eacute;t&eacute; reconnue comme jouant un r&ocirc;le crucial en d&eacute;fense cellulaire contre des conditions stressantes, non seulement li&eacute;es &agrave; la lib&eacute;ration de l&#39;h&egrave;me. HO-1 prot&egrave;ge l&#39;apoptose des cellules endoth&eacute;liales, est impliqu&eacute; dans la relaxation de vaisseaux sanguins r&eacute;glementant le tonus vasculaire, att&eacute;nue la r&eacute;ponse inflammatoire dans la paroi vasculaire et participe &agrave; la formation de vaisseaux sanguins au moyen de l&#39;angiogen&egrave;se et vasculog&eacute;n&egrave;se. Les fonctions de ce dernier lien HO-1 non seulement &agrave; l&#39;isch&eacute;mie cardiovasculaire, mais aussi &agrave; d&#39;autres affections, qui, comme le d&eacute;veloppement, la cicatrisation des plaies ou cancer, d&eacute;pendent de la n&eacute;ovascularisation. Le but de cette &eacute;tude est d&#39;aborder les m&eacute;canismes de r&eacute;gulation HO-1 et la fonction dans la pathologie et la physiologie cardio-vasculaire et de montrer quelques applications possibles des vastes connaissances produites jusqu&#39;&agrave; maintenant. Des donn&eacute;es r&eacute;centes prouvent puissant pour l&#39;implication de HO-1 dans l&#39;effet th&eacute;rapeutique des m&eacute;dicaments utilis&eacute;s dans les maladies cardiovasculaires. Nouvelles &eacute;tudes ouvrent les possibilit&eacute;s d&#39;application de HO-1 pour la th&eacute;rapie g&eacute;nique et cellulaire. Par cons&eacute;quent, recherche dans les prochaines ann&eacute;es devrait aider &agrave; &eacute;lucider le r&ocirc;le r&eacute;el des HO-1 dans l&#39;effet des drogues et de la faisabilit&eacute; clinique de th&eacute;rapie cellulaire et g&eacute;nique HO-1-bas&eacute;, cr&eacute;ant les avenues th&eacute;rapeutiques efficaces pour cela raffin&eacute; syst&egrave;me antioxydant.

H&egrave;me oxyg&eacute;nase-1 et le lit vasculaire : des m&eacute;canismes mol&eacute;culaires des possibilit&eacute;s th&eacute;rapeutiques.

H&auml;m-Oxygenase-1, ein Enzym, die erniedrigende H&auml;m und Kohlenmonoxid, Eisen und Biliverdin, ist erkannt worden, als spielen eine entscheidende Rolle in der zellul&auml;ren Abwehr gegen Stress Bedingungen, nicht nur f&uuml;r die H&auml;m-Version. HO-1 endotheliale Zellen vor der Apoptose sch&uuml;tzt, Blutgef&auml;&szlig; Entspannung regulieren Gef&auml;&szlig;tonus beteiligt ist, d&auml;mpft Entz&uuml;ndungsreaktion in der Gef&auml;&szlig;wand und beteiligt sich an Blutgef&auml;&szlig; Bildung durch Angiogenese und Vaskulogenese. Die letztgenannten Funktionen verkn&uuml;pfen HO-1 nicht nur Herz-Kreislauf-Isch&auml;mie, sondern auch viele andere Bedingungen, die wie Entwicklung, Wundheilung oder Krebs, Neovaskularisation abh&auml;ngig sind. Diese umfassende &Uuml;berpr&uuml;fung zielt, die Mechanismen der HO-1 Verordnung und Funktion in der Herz-Kreislauf-Physiologie und Pathologie anzusprechen und einige Einsatzm&ouml;glichkeiten von riesigen wissen bisher zu demonstrieren. Aktuelle Daten liefern starken Beweise f&uuml;r die Beteiligung der HO-1 in die therapeutische Wirkung der Medikamente bei Herz-Kreislauf-Krankheiten. Neuartige Studien &ouml;ffnen die Anwendungsm&ouml;glichkeiten der HO-1 f&uuml;r gen- und Therapie. Daher Forschung in den kommenden Jahren soll dazu beitragen, sowohl die reale Rolle der HO-1 in die Wirkung von Drogen und die klinischen Durchf&uuml;hrbarkeit der HO-1-basierte Zelle und gen-Therapie, erhellen erstellen die effektiven therapeutischen M&ouml;glichkeiten f&uuml;r diese antioxidativen System verfeinert.

H&auml;m-Oxygenase-1 und das vaskul&auml;re Bett: von molekularen Mechanismen zu therapeutischen M&ouml;glichkeiten.

Eme ossigenasi-1, un enzima degradante eme di monossido di carbonio, ferro e Biliverdina, &egrave; stato riconosciuto come giocare un ruolo cruciale nella difesa cellulare contro condizioni stressanti, non solo relative al rilascio di EME. HO-1 protegge le cellule endoteliali dal apoptosis, &egrave; coinvolto nel rilassamento di vasi sanguigni che regolano il tono vascolare, attenua la risposta infiammatoria nella parete del vaso e partecipa alla formazione di vasi sanguigni mediante angiogenesi e vasculogenesi. Le funzioni di quest&#39;ultime link HO-1 non solo per ischemia cardiovascolare, ma anche per molte altre condizioni che, come lo sviluppo, la guarigione della ferita o il cancro, sono dipendente da neovascolarizzazione. L&#39;obiettivo di questa revisione globale &egrave; di affrontare i meccanismi di regolamento HO-1 e la funzione cardiovascolare, fisiologia e patologia e per dimostrare alcune possibili applicazioni della vasta conoscenza generato finora. Dati recenti forniscono potenti prove del coinvolgimento di HO-1 nell&#39;effetto terapeutico dei farmaci utilizzati nelle malattie cardiovascolari. Studi romanzo aprire le possibilit&agrave; di applicazione di HO-1 per la terapia genica e cellulare. Quindi, ricerca nei prossimi anni dovrebbe aiutare a chiarire sia il vero ruolo di HO-1 l&#39;effetto di droghe e la fattibilit&agrave; clinica della terapia cellulare e genica basate su HO-1, creando viali terapeutici efficaci per questo raffinato sistema antiossidante.

Eme ossigenasi-1 e il letto vascolare: dai meccanismi molecolari di opportunit&agrave; terapeutiche.

&#x30D8;&#x30E0;&#x9178;&#x7D20;&#x6DFB;&#x52A0;&#x9175;&#x7D20;-1&#x3001;&#x4E00;&#x9178;&#x5316;&#x70AD;&#x7D20;&#x3001;&#x9244;&#x3001;&#x304A;&#x3088;&#x3073;&#x30D3;&#x30EA;&#x30D9;&#x30EB;&#x30B8;&#x30F3;&#x3001;&#x30D8;&#x30E0;&#x5206;&#x89E3;&#x9175;&#x7D20;&#x306F;&#x3001;&#x30D8;&#x30E0;&#x306E;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x306B;&#x95A2;&#x9023;&#x3059;&#x308B;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x3001;&#x30B9;&#x30C8;&#x30EC;&#x30B9;&#x306E;&#x591A;&#x3044;&#x72B6;&#x6CC1;&#x306B;&#x5BFE;&#x3059;&#x308B;&#x7D30;&#x80DE;&#x306E;&#x9632;&#x5FA1;&#x306B;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x3092;&#x518D;&#x751F;&#x3059;&#x308B;&#x3068;&#x3057;&#x3066;&#x8A8D;&#x8B58;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;HO-1 &#x306F;&#x3001;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x304C;&#x30A2;&#x30DD;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x304B;&#x3089;&#x4FDD;&#x8B77;&#x3059;&#x308B;&#x8840;&#x7BA1;&#x7DCA;&#x5F35;&#x3092;&#x898F;&#x5236;&#x3059;&#x308B;&#x8840;&#x7BA1;&#x5F1B;&#x7DE9;&#x306B;&#x95A2;&#x4E0E;&#x3057;&#x3066;&#x3044;&#x308B;&#x3001;&#x8840;&#x7BA1;&#x58C1;&#x306E;&#x708E;&#x75C7;&#x5FDC;&#x7B54;&#x306E;&#x6E1B;&#x8870;&#x3057;&#x3001;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3068;&#x8840;&#x7BA1;&#x306B;&#x3088;&#x308B;&#x8840;&#x7BA1;&#x5F62;&#x6210;&#x306B;&#x53C2;&#x52A0;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x5F8C;&#x8005;&#x306E;&#x6A5F;&#x80FD;&#x306F; HO 1 &#x5FC3;&#x8840;&#x7BA1;&#x865A;&#x8840;&#x306E;&#x307F;&#x306A;&#x3089;&#x305A;&#x3001;&#x958B;&#x767A;&#x3001;&#x5275;&#x50B7;&#x6CBB;&#x7652;&#x3001;&#x307E;&#x305F;&#x306F;&#x764C;&#x306E;&#x3088;&#x3046;&#x306A;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x306B;&#x4F9D;&#x5B58;&#x3059;&#x308B;&#x4ED6;&#x306E;&#x591A;&#x304F;&#x306E;&#x6761;&#x4EF6;&#x306B;&#x3082;&#x30EA;&#x30F3;&#x30AF;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x5305;&#x62EC;&#x7684;&#x306A;&#x898B;&#x76F4;&#x3057;&#x306E;&#x76EE;&#x7684;&#x306F;&#x3001;HO 1 &#x898F;&#x5236;&#x3068;&#x5FC3;&#x8840;&#x7BA1;&#x7CFB;&#x306E;&#x751F;&#x7406;&#x3068;&#x75C5;&#x7406;&#x3067;&#x95A2;&#x6570;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x306B;&#x5BFE;&#x51E6;&#x3059;&#x308B;&#x305F;&#x3081;&#x3053;&#x308C;&#x307E;&#x3067;&#x81A8;&#x5927;&#x306A;&#x77E5;&#x8B58;&#x306E;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E;&#x53EF;&#x80FD;&#x306A;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x793A;&#x3059;&#x3053;&#x3068;&#x3067;&#x3059;&#x3002;&#x6700;&#x8FD1;&#x306E;&#x30C7;&#x30FC;&#x30BF;&#x306F; HO 1 &#x306E;&#x95A2;&#x4E0E;&#x3067;&#x5FC3;&#x8840;&#x7BA1;&#x75BE;&#x60A3;&#x306B;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x308B;&#x85AC;&#x5264;&#x306E;&#x6CBB;&#x7642;&#x52B9;&#x679C;&#x306E;&#x5F37;&#x529B;&#x306A;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;&#x5C0F;&#x8AAC;&#x306E;&#x7814;&#x7A76;&#x3078;&#x306E; HO 1 &#x907A;&#x4F1D;&#x5B50;&#x7D30;&#x80DE;&#x6CBB;&#x7642;&#x306E;&#x5FDC;&#x7528;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x958B;&#x304D;&#x307E;&#x3059;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;&#x4ECA;&#x5F8C;&#x5E74;&#x9593;&#x3067;&#x7814;&#x7A76; HO-1 &#x85AC;&#x7269;&#x306E;&#x52B9;&#x679C;&#x3068; HO 1 &#x306B;&#x57FA;&#x3065;&#x3044;&#x3066;&#x3001;&#x7D30;&#x80DE;&#x3001;&#x907A;&#x4F1D;&#x5B50;&#x6CBB;&#x7642;&#x306E;&#x81E8;&#x5E8A;&#x7684;&#x53EF;&#x80FD;&#x6027;&#x306E;&#x4E21;&#x65B9;&#x5B9F;&#x306E;&#x5F79;&#x5272;&#x3092;&#x89E3;&#x660E;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x5F79;&#x7ACB;&#x3064;&#x306F;&#x305A;&#x3053;&#x306E;&#x305F;&#x3081;&#x306E;&#x52B9;&#x679C;&#x7684;&#x306A;&#x6CBB;&#x7642;&#x624B;&#x6BB5;&#x3092;&#x4F5C;&#x6210;&#x6D17;&#x7DF4;&#x3055;&#x308C;&#x305F;&#x6297;&#x9178;&#x5316;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3002;

&#x30D8;&#x30E0; &#x30D8;&#x30E0;&#x30AA;&#x30AD;&#x30B7;&#x30B2;&#x30CA;&#x30FC;&#x30BC; &#x2010; 1 &#x304A;&#x3088;&#x3073;&#x8840;&#x7BA1;&#x5E8A;: &#x6CBB;&#x7642;&#x306E;&#x6A5F;&#x4F1A;&#x306B;&#x5206;&#x5B50;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x304B;&#x3089;&#x3002;

Heme oxygenase-1, &#xC77C;&#xC0B0;&#xD654; &#xD0C4;&#xC18C;, &#xCCA0;, biliverdin, heme &#xC800;&#xD558; &#xD6A8;&#xC18C; heme &#xB9B4;&#xB9AC;&#xC2A4; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; &#xAD00;&#xB828; &#xB41C; &#xC2A4;&#xD2B8;&#xB808;&#xC2A4;&#xAC00; &#xB9CE;&#xC740; &#xC870;&#xAC74;&#xC5D0; &#xB300; &#xD55C; &#xC138;&#xD3EC; &#xBC29;&#xC5B4;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xC5ED;&#xD560;&#xC744; &#xC778;&#xC815;&#xC744; &#xBC1B;&#xACE0; &#xC788;&#xB2E4;. &#xD638; 1 apoptosis&#xC5D0;&#xC11C; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xB97C; &#xBCF4;&#xD638; &#xD558; &#xD608;&#xAD00; &#xD1A4; &#xC870;&#xC808;-&#xD608;&#xAD00; &#xC774;&#xC644;&#xC5D0; &#xAD00;&#xC5EC; &#xD558; &#xACE0; &#xD608;&#xAD00; &#xBCBD;&#xC758; &#xC5FC;&#xC99D; &#xBC18;&#xC751;&#xC744; &#xC57D;&#xD654; &#xD608;&#xAD00; &#xC2E0;&#xC0DD;&#xACFC; vasculogenesis&#xC5D0; &#xC758;&#xD574;-&#xD608;&#xAD00; &#xD615;&#xC131;&#xC5D0; &#xCC38;&#xC5EC; &#xD558;. &#xD6C4;&#xC790;&#xC758; &#xAE30;&#xB2A5; &#xB9C1;&#xD06C; &#xD638; 1 &#xC2EC;&#xD608; &#xAD00;&#xACC4; &#xD5C8; &#xD608; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; &#xADF8; &#xAC1C;&#xBC1C;, &#xC0C1;&#xCC98; &#xCE58;&#xC720;, &#xB610;&#xB294; &#xC554;&#xACFC; &#xAC19;&#xC740; neovascularization&#xC5D0; &#xC758;&#xC874; &#xD558;&#xB294; &#xB2E4;&#xB978; &#xB9CE;&#xC740; &#xC870;&#xAC74;&#xC5D0;. &#xC774; &#xD3EC;&#xAD04;&#xC801;&#xC778; &#xAC80;&#xD1A0;&#xC758; &#xBAA9;&#xC801;&#xC740; &#xD638;-1 &#xB808; &#xADE4; &#xB808;&#xC774; &#xC158;&#xACFC; &#xC2EC;&#xD608; &#xAD00; &#xC0DD;&#xB9AC; &#xBC0F; &#xBCD1; &#xB9AC; &#xAE30;&#xB2A5; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC744; &#xD574;&#xACB0; &#xD558; &#xACE0; &#xC9C0;&#xAE08;&#xAE4C;&#xC9C0; &#xC0DD;&#xC131; &#xB41C; &#xAD11;&#xB300; &#xD55C; &#xC9C0;&#xC2DD;&#xC758; &#xBA87; &#xAC00;&#xC9C0; &#xAC00;&#xB2A5;&#xD55C; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;&#xC744; &#xBCF4;&#xC5EC;. &#xCD5C;&#xADFC; &#xB370;&#xC774;&#xD130;&#xB294; &#xC2EC;&#xD608; &#xAD00;&#xACC4; &#xC9C8;&#xD658;&#xC5D0; &#xC0AC;&#xC6A9; &#xB418;&#xB294; &#xC57D;&#xBB3C;&#xC758; &#xCE58;&#xB8CC; &#xD6A8;&#xACFC;&#xC5D0; &#xD638; 1&#xC758; &#xCC38;&#xC5EC;&#xC5D0; &#xB300; &#xD55C; &#xAC15;&#xB825;&#xD55C; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xC18C;&#xC124; &#xC5F0;&#xAD6C; &#xC720;&#xC804;&#xC790; &#xBC0F; &#xC138;&#xD3EC; &#xCE58;&#xB8CC;&#xC5D0; &#xB300; &#xD55C; &#xD638;-1&#xC758; &#xC801;&#xC6A9; &#xAC00;&#xB2A5;&#xC131;&#xC744; &#xC5FD;&#xB2C8;&#xB2E4;. &#xB530;&#xB77C;&#xC11C; &#xCC28;&#xAE30; &#xB144;&#xC5D0;&#xC11C; &#xC5F0;&#xAD6C; &#xD574;&#xC57C; &#xB450; &#xC57D;&#xBB3C;&#xC758; &#xD6A8;&#xACFC;&#xC640; &#xD638; 1 &#xAE30;&#xBC18; &#xC138;&#xD3EC; &#xBC0F; &#xC720;&#xC804;&#xC790; &#xCE58;&#xB8CC;&#xC758; &#xC784;&#xC0C1; &#xD0C0;&#xB2F9;&#xC131; &#xD638;-1&#xC758; &#xC2E4;&#xC81C; &#xC5ED;&#xD560;&#xC744; &#xBA85;&#xB8CC; &#xD558; &#xAC8C; &#xB3C4;&#xC6C0;&#xC774; &#xB420;&#xC774; &#xB300; &#xD55C; &#xD6A8;&#xACFC;&#xC801;&#xC778; &#xCE58;&#xB8CC; &#xC218;&#xB2E8;&#xC744; &#xB9CC;&#xB4DC;&#xB294; &#xC138;&#xB828; &#xB41C; &#xD56D; &#xC0B0;&#xD654; &#xC2DC;&#xC2A4;&#xD15C;.

Heme oxygenase-1&#xACFC; &#xD608;&#xAD00; &#xCE68;&#xB300;: &#xCE58;&#xB8CC; &#xAE30;&#xD68C;&#xB97C; &#xBD84;&#xC790; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC5D0;&#xC11C;.

Hemo oxigenasa-1, una enzima degradante heme a mon&oacute;xido de carbono, hierro y biliverdina, ha sido reconocido como desempe&ntilde;ando un papel crucial en la defensa celular contra condiciones estresantes, no s&oacute;lo relacionados con la liberaci&oacute;n de Hem. HO-1 protege a las c&eacute;lulas endoteliales de apoptosis, participa en la relajaci&oacute;n de los vasos sangu&iacute;neos, regular el tono vascular, aten&uacute;a la respuesta inflamatoria en la pared del vaso y participa en la formaci&oacute;n de vasos sangu&iacute;neos por medio de la angiog&eacute;nesis y vasculog&eacute;nesis. Las funciones de este &uacute;ltimas enlazan HO-1 no s&oacute;lo a la isquemia cardiovascular, sino tambi&eacute;n a muchas otras condiciones que, como desarrollo, la cicatrizaci&oacute;n de heridas o c&aacute;ncer, dependen de la neovascularizaci&oacute;n. El objetivo de esta revisi&oacute;n completa es abordar los mecanismos de regulaci&oacute;n de HO-1 y funci&oacute;n en la fisiolog&iacute;a cardiovascular y patolog&iacute;a y para demostrar algunas posibles aplicaciones del vasto conocimiento generado hasta ahora. Recientes datos proporcionan evidencia poderosa para la participaci&oacute;n de HO-1 en el efecto terap&eacute;utico de los f&aacute;rmacos utilizados en las enfermedades cardiovasculares. Nuevos estudios abierta las posibilidades de aplicaci&oacute;n de HO-1 para la terapia g&eacute;nica y celular. Por lo tanto, la investigaci&oacute;n en los pr&oacute;ximos a&ntilde;os debe ayudar a aclarar ambos el papel real de HO-1 en el efecto de f&aacute;rmacos y la viabilidad cl&iacute;nica de HO-1-basados en terapia celular y g&eacute;nica, creando las v&iacute;as terap&eacute;uticas eficaces para este refinado sistema antioxidante.

Hemo oxigenasa-1 y el lecho vascular: de mecanismos moleculares para oportunidades terap&eacute;uticas.

5T4 oncofetal molecules are highly expressed during development and upregulated in cancer while showing only low levels in some adult tissues. Upregulation of 5T4 expression is a marker of loss of pluripotency in the early differentiation of embryonic stem (ES) cells and forms an integrated component of an epithelial-mesenchymal transition, a process important during embryonic development and metastatic spread of epithelial tumors. Investigation of the transcriptional changes in early ES differentiation showed upregulation of CXCL12 and down-regulation of a cell surface protease, CD26, which cleaves this chemokine. CXCL12 binds to the widely expressed CXCR4 and regulates key aspects of development, stem cell motility and tumour metastasis to tissues with high levels of CXCL12. We show that the 5T4 glycoprotein is required for optimal functional cell surface expression of the chemokine receptor CXCR4 and CXCL12 mediated chemotaxis in differentiating murine embryonic stem cells and embryo fibroblasts (MEF). Cell surface expression of 5T4 and CXCR4 molecules is co-localized in differentiating ES cells and MEF. By contrast, differentiating ES and MEF derived from 5T4 knockout (KO) mice show only intracellular CXCR4 expression but infection with adenovirus encoding mouse 5T4 restores CXCL12 chemotaxis and surface co-localization with 5T4 molecules. A series of chimeric constructs with interchanged domains of 5T4 and the glycoprotein CD44 were used to map the 5T4 sequences relevant for CXCR4 membrane expression and function in 5T4KO MEF. These data identified the 5T4 transmembrane domain as sufficient and necessary to enable CXCR4 cell surface expression and chemotaxis. Furthermore, some monoclonal antibodies against m5T4 can inhibit CXCL12 chemotaxis of differentiating ES cells and MEF which is not mediated by simple antigenic modulation. Collectively, these data support a molecular interaction of 5T4 and CXCR4 occurring at the cell surface which directly facilitates the biological response to CXCL12. The regulation of CXCR4 surface expression by 5T4 molecules is a novel means to control responses to the chemokine CXCL12 for example during embryogenesis but can also be selected to advantage the spread of a 5T4 positive tumor from its primary site.

CXCR4 mediated chemotaxis is regulated by 5T4 oncofetal glycoprotein in mouse embryonic cells.

Bone marrow-derived cells have been postulated as a source of multipotent mesenchymal stem cells (MSC). However, the whole fraction of MSC remains heterogeneous and the expansion of primitive subset of these cells is still not well established. Here, we optimized the protocol for propagating the low-adherent subfraction of MSC which results in long-term expansion of population characterized by CD45(-)CD14(+)CD34(+) phenotype along with expression of common MSC markers. We established that the expanded MSC are capable of differentiating into endothelial cells highly expressing angiogenic markers and exhibiting functional properties of endothelium. Moreover, we found these cells to be multipotent and capable of giving rise into cells from neuronal lineages. Interestingly, the expanded MSC form characteristic cellular spheres in vitro indicating primitive features of these cells. In sum, we isolated the novel multipotent subpopulation of CD45(-)CD14(+) CD34(+) bone marrow-derived cells that could be maintained in long-term culture without losing this potential.

CD45-CD14 +CD34 + murine bone marrow low-adherent mesenchymal primitive cells preserve multilineage differentiation potential in long-term in vitro culture.

Effective vaccination against tumour-associated antigens (TAA) such as the 5T4 oncofoetal glycoprotein may be limited by the nature of the T cell repertoire and the influence of immunomodulatory factors in particular T regulatory cells (Treg). Here, we identified mouse 5T4-specific T cell epitopes using a 5T4 knock out (5T4KO) mouse and evaluated corresponding wild-type (WT) responses as a model to refine and improve immunogenicity. We have shown that 5T4KO mice vaccinated by replication defective adenovirus encoding mouse 5T4 (Adm5T4) generate potent 5T4-specific IFN-Î³ CD8 and CD4 T cell responses which mediate significant protection against 5T4 positive tumour challenge. 5T4KO CD8 but not CD4 primed T cells also produced IL-17. By contrast, Adm5T4-immunized WT mice showed no tumour protection consistent with only low avidity CD8 IFN-Î³, no IL-17 T cell responses and no detectable CD4 T cell effectors producing IFN-Î³ or IL-17. Treatment with anti-folate receptor 4 (FR4) antibody significantly reduced the frequency of Tregs in WT mice and enhanced 5T4-specific IFN-Î³ but reduced IL-10 T cell responses but did not reveal IL-17-producing effectors. This altered balance of effectors by treatment with FR4 antibody after Adm5T4 vaccination provided modest protection against autologous B16m5T4 melanoma challenge. The efficacy of 5T4 and some other TAA vaccines may be limited by the combination of TAA-specific T regs, the deletion and/or alternative differentiation of CD4 T cells as well as the absence of distinct subsets of CD8 T cells.

Regulation of autologous immunity to the mouse 5T4 oncofoetal antigen: implications for immunotherapy.

Interferons (IFN) play a pivotal role in innate immunity, orchestrating a cell-intrinsic anti-pathogenic state and stimulating adaptive immune responses. The complex interplay between the primary response to IFNs and its modulation by positive and negative feedback loops is incompletely understood. Here, we implement the combination of high-resolution gene-expression profiling of nascent RNA with translational inhibition of secondary feedback by cycloheximide. Unexpectedly, this approach revealed a prominent role of negative feedback mechanisms during the immediate (â‰¤60 min) IFNÎ± response. In contrast, a more complex picture involving both negative and positive feedback loops was observed on IFNÎ³ treatment. IFNÎ³-induced repression of genes associated with regulation of gene expression, cellular development, apoptosis and cell growth resulted from cycloheximide-resistant primary IFNÎ³ signalling. In silico promoter analysis revealed significant overrepresentation of SP1/SP3-binding sites and/or GC-rich stretches. Although signal transducer and activator of transcription 1 (STAT1)-binding sites were not overrepresented, repression was lost in absence of STAT1. Interestingly, basal expression of the majority of these IFNÎ³-repressed genes was dependent on STAT1 in IFN-naÃ¯ve fibroblasts. Finally, IFNÎ³-mediated repression was also found to be evident in primary murine macrophages. IFN-repressed genes include negative regulators of innate and stress response, and their decrease may thus aid the establishment of a signalling perceptive milieu.

Deciphering the modulation of gene expression by type I and II interferons combining 4sU-tagging, translational arrest and in silico promoter analysis.

Andrzej J. Rutkowski

Department of Medicine

University of Cambridge

Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture

Andrzej J. Rutkowski

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Medicine

University of Cambridge

Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture

Department of Medicine