molecular-profiling-invasive-tumor-microenvironment-3-dimensional

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and <em>Ex vivo</em> Fibroblasts

Invading colorectal cancer (CRC) cells have acquired the capacity to break free from their sister cells, infiltrate the stroma, and remodel the ...

London Research Institute, Cancer Research UK

&#x6211;&#x4EEC;&#x6700;&#x8FD1;&#x5B8C;&#x6210;&#x4E86; spiruchostatin A&#xFF0C;depsipeptide &#x4E0E;&#x7ED3;&#x6784;&#x76F8;&#x4F3C;&#xFF0C;&#x5230; FK228&#xFF0C;&#x4E00;&#x79CD;&#x7EC4;&#x86CB;&#x767D;&#x53BB;&#x4E59;&#x9170;&#x9176; (HDAC) &#x6291;&#x5236; (HDI) &#x76EE;&#x524D;&#x6B63;&#x5728;&#x8BC4;&#x4F30;&#x5728;&#x764C;&#x75C7;&#x7684;&#x4E34;&#x5E8A;&#x8BD5;&#x9A8C;&#x4E2D;&#x7684;&#x5929;&#x7136;&#x4EA7;&#x7269;&#x7684;&#x5168;&#x5408;&#x6210;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x62A5;&#x544A;&#x8BE6;&#x7EC6;&#x7684;&#x7279;&#x6027; spiruchostatin A.Spiruchostatin A &#x662F;&#x4E00;&#x79CD;&#x70C8;&#x6027;&#x7684; &#xFF08;sub nM&#xFF09; &#x6291;&#x5236;&#x5242;&#x7684;&#x4F53;&#x5916;&#x6D3B;&#x6027; I &#x7C7B; HDAC &#x6D3B;&#x6027;&#x4F53;&#x5916;&#xFF0C;&#x5E76;&#x4F5C;&#x4E3A;&#x524D;&#x4F53;&#x836F;&#x7269;&#xFF0C;&#x9700;&#x8981;&#x51CF;&#x5C11;&#x6D3B;&#x52A8;&#x91C7;&#x53D6;&#x884C;&#x52A8;&#x3002;Spiruchostatin A &#x662F;&#x751F;&#x957F;&#x7684;&#x4E00;&#x79CD;&#x70C8;&#x6027; &#xFF08;&#x4F4E; nM&#xFF09; &#x6291;&#x5236;&#x5242;&#x5404;&#x79CD;&#x764C;&#x7EC6;&#x80DE;&#x3002;Spiruchostatin A &#x8BF1;&#x53D1;&#x5185;&#x7EC4;&#x86CB;&#x767D; H3 &#x548C; H4&#xFF0C;&#x5177;&#x4F53;&#x8D56;&#x6C28;&#x9178;&#x7684;&#x4E59;&#x9170;&#x5316;&#x548C;&#x589E;&#x52A0;&#x8868;&#x8FBE;&#x7684; p21(cip1/waf1)&#xFF0C;&#x4F46;&#x786E;&#x5B9E;&#x4E0D;&#x4FC3;&#x4F7F; &alpha;-&#x5FAE;&#x7BA1;&#x86CB;&#x767D;&#x7684;&#x4E59;&#x9170;&#x5316;&#x3002;Spiruchostatin A &#x8FD8;&#x5728; MCF7 &#x4E73;&#x764C;&#x7EC6;&#x80DE;&#x8BF1;&#x5BFC;&#x7EC6;&#x80DE;&#x5468;&#x671F;&#x902E;&#x6355;&#x3001; &#x5206;&#x5316;&#x548C;&#x7EC6;&#x80DE;&#x6B7B;&#x4EA1;&#x3002;&#x50CF; FK228&#xFF0C;spiruchostatin A &#x662F;&#x8BF1;&#x5BFC;&#x56E0;&#x5B50;&#x548C; ABCB1 &#x836F;&#x7269;&#x5916;&#x6392;&#x6CF5;&#x7684;&#x886C;&#x5E95;&#x3002;&#x800C; spiruchostatin A &#x548C; FK228 &#x8BF1;&#x5BFC;&#x65F7;&#x65E5;&#x6301;&#x4E45;&#x7684;&#x7EC4;&#x86CB;&#x767D;&#x4E59;&#x9170;&#x5316;&#x4F5C;&#x7528;&#xFF0C;&#x809F; HDI &#x81F4;&#x77ED;&#x547D;&#x7EC4;&#x86CB;&#x767D;&#x4E59;&#x9170;&#x5316;&#x3002;&#x4F7F;&#x7528;&#x786E;&#x5B9A;&#x7684;&#x57FA;&#x56E0;&#x82AF;&#x7247;&#x5206;&#x6790;&#x7684;&#x4EBA;&#x7C7B;&#x53D1;&#x5C55;&#x6307;&#x6570;&#x76EE;&#x6807;&#x57FA;&#x56E0;&#x7684;&#x4E00;&#x4E2A;&#x5B50;&#x96C6;&#xFF0C;&#x6211;&#x4EEC;&#x8868;&#x660E;&#x8FD9;&#x4E9B;&#x5DEE;&#x5F02;&#x5728;&#x4EBA;&#x7C7B;&#x53D1;&#x5C55;&#x6307;&#x6570;&#x4E4B;&#x95F4;&#x7684;&#x7EC4;&#x86CB;&#x767D;&#x4E59;&#x9170;&#x5316;&#x52A8;&#x529B;&#x5B66;&#x4E0E;&#x52A8;&#x529B;&#x5B66;&#x7684;&#x611F;&#x5E94;&#x6216;&#x9547;&#x538B;&#x7684;&#x7279;&#x5B9A;&#x76EE;&#x6807;&#x57FA;&#x56E0;&#x7684;&#x5DEE;&#x5F02;&#x76F8;&#x5173;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x8868;&#x660E;&#xFF0C;spiruchostatin A &#x662F;&#x4E00;&#x79CD;&#x5F3A;&#x6548;&#x6291;&#x5236;&#x5242;&#x7C7B;&#x6211; HDACs &#x548C;&#x6297;&#x764C;&#x5242;&#x3002;&#x52A8;&#x529B;&#x5B66;&#x7814;&#x7A76;&#x7684;&#x4EBA;&#x7C7B;&#x53D1;&#x5C55;&#x5021;&#x8BAE;&#x884C;&#x52A8;&#x7684;&#x5DEE;&#x5F02;&#x53EF;&#x80FD;&#x662F;&#x91CD;&#x8981;&#x7684;&#x662F;&#x8FD9;&#x4E9B;&#x5316;&#x5408;&#x7269;&#x7684;&#x4E34;&#x5E8A;&#x5E94;&#x7528;&#x3002;

Depsipeptide &#x7EC4;&#x86CB;&#x767D;&#x53BB;&#x4E59;&#x9170;&#x9176;&#x6291;&#x5236;&#x5242; spiruchostatin A.&#x4F53;&#x5916;&#x6D3B;&#x6027;&#x7684;&#x7279;&#x6027;&#x63CF;&#x8FF0;

We recently completed the total synthesis of spiruchostatin A, a depsipeptide natural product with close structural similarities to FK228, a histone deacetylase (HDAC) inhibitor (HDI) currently being evaluated in clinical trials for cancer. Here we report a detailed characterisation of the in vitro activity of spiruchostatin A. Spiruchostatin A was a potent (sub-nM) inhibitor of class I HDAC activity in vitro and acted as a prodrug, requiring reduction for activity. Spiruchostatin A was a potent (low nM) inhibitor of the growth of various cancer cell lines. Spiruchostatin A-induced acetylation of specific lysine residues within histones H3 and H4, and increased the expression of p21(cip1/waf1), but did not induce acetylation of alpha-tubulin. Spiruchostatin A also induced cell cycle arrest, differentiation and cell death in MCF7 breast cancer cells. Like FK228, spiruchostatin A was both an inducer and substrate of the ABCB1 drug efflux pump. Whereas spiruchostatin A and FK228-induced protracted histone acetylation, hydroxamate HDI-induced short-lived histone acetylation. Using a subset of HDI-target genes identified by microarray analysis, we demonstrated that these differences in kinetics of histone acetylation between HDI correlated with differences in the kinetics of induction or repression of specific target genes. Our results demonstrate that spiruchostatin A is a potent inhibitor of class I HDACs and anti-cancer agent. Differences in the kinetics of action of HDI may be important for the clinical application of these compounds.

Characterisation of the in vitro activity of the depsipeptide histone deacetylase inhibitor spiruchostatin A.

R&eacute;cemment, nous avons termin&eacute; la synth&egrave;se totale du spiruchostatin A, un produit naturel depsipeptide &eacute;troites similitudes structurelles &agrave; FK228, histone d&eacute;sac&eacute;tylase (HDAC) inhibiteur (IDH) en cours d&#39;&eacute;valuation dans des essais cliniques pour le cancer. Nous rapportons ici une caract&eacute;risation d&eacute;taill&eacute;e de l&#39;activit&eacute; in vitro de spiruchostatin A. Spiruchostatin A est un inhibiteur puissant (sub-nM) de l&#39;activit&eacute; in vitro et a agi comme une prodrogue, n&eacute;cessitant une r&eacute;duction d&#39;activit&eacute; HDAC de classe I. Spiruchostatin A est un inhibiteur puissant (faible nM) de la croissance de diverses lign&eacute;es cellulaires canc&eacute;reuses. L&#39;ac&eacute;tylation des r&eacute;sidus lysine sp&eacute;cifique au sein des histones H3 et H4, induite par le Spiruchostatin A augment&eacute; l&#39;expression de p21(cip1/waf1), et n&#39;a pas induit d&#39;ac&eacute;tylation des alpha-tubuline. Spiruchostatin A &eacute;galement provoqu&eacute; mort cellulaire, la diff&eacute;renciation et l&#39;arrestation de cycle de cellules en cellules de cancer du sein MCF7. Comme FK228, spiruchostatin A &eacute;tait un inducteur et le substrat de la pompe &agrave; efflux ABCB1 drogue. Consid&eacute;rant que spiruchostatin A et l&#39;ac&eacute;tylation des histones prolong&eacute;e induite par le FK228, l&#39;ac&eacute;tylation des histones de courte dur&eacute;e induite par l&#39;IDH hydroxamate. &Agrave; l&#39;aide d&#39;un sous-ensemble de g&egrave;nes HDI-cibles identifi&eacute;es par analyse microarray, nous avons d&eacute;montr&eacute; que ces diff&eacute;rences de cin&eacute;tique de l&#39;ac&eacute;tylation des histones entre HDI en corr&eacute;lation avec les diff&eacute;rences dans la cin&eacute;tique de l&#39;induction ou r&eacute;pression des g&egrave;nes cibles sp&eacute;cifiques. Nos r&eacute;sultats d&eacute;montrent que spiruchostatin A est un puissant inhibiteur de la classe j&#39;ai HDACs et agent anticanc&eacute;reux. Diff&eacute;rences dans la cin&eacute;tique d&#39;action du HDI peuvent &ecirc;tre importants pour l&#39;application clinique de ces compos&eacute;s.

Caract&eacute;risation de l&#39;activit&eacute; in vitro de la depsipeptide histone deacetylase inhibiteur spiruchostatin A.

Wir vor kurzem die Totalsynthese von Spiruchostatin A, ein Depsipeptide Naturprodukt mit enge strukturelle &Auml;hnlichkeiten zu FK228, ein Histon-Deacetylase (HDAC)-Inhibitor (HDI) derzeit in klinischen Studien f&uuml;r Krebs ausgewertet. Hier berichten wir eine detaillierte Charakterisierung der in-vitro-Aktivit&auml;t von Spiruchostatin A. Spiruchostatin A ein potenter (Sub-nM)-Inhibitor der war Klasse I HDAC-Aktivit&auml;t in vitro und gehandelt als ein Prodrug, Erm&auml;&szlig;igung f&uuml;r Aktivit&auml;t erfordern. Spiruchostatin A war ein potenter (niedrige nM)-Inhibitor, der das Wachstum der verschiedenen Krebs-Zelllinien. Spiruchostatin A-induzierte Acetylierung der spezifischen Lysin R&uuml;ckst&auml;nde innerhalb Histone H3 und H4, und erh&ouml;ht den Ausdruck des p21(cip1/waf1), aber nicht induzieren Acetylierung des Alpha-Tubulin. Spiruchostatin A induzierten Zellzyklus Verhaftung, Differenzierung und Zelle Tod auch in Brustkrebs-Zellen MCF7. Wie FK228 war Spiruchostatin A Induktor sowohl Substrat der ABCB1 Droge Efflux Pumpe. W&auml;hrend die Spiruchostatin A und FK228-induzierte langwierigen Histon Acetylierung, Hydroxamate HDI-induzierte kurzlebige Histon-Acetylierung. Verwenden eine Teilmenge der HDI-Zielgene von Microarray-Analyse identifiziert, wir gezeigt, dass diese Unterschiede in der Kinetik der Histon-Acetylierung zwischen HDI mit Unterschieden in der Kinetik der Induktion oder Repression spezifische Zielgene korreliert. Unsere Ergebnisse zeigen, dass Spiruchostatin A ist ein potenter Inhibitor der Klasse I HDACs und Krebswirkstoff. Unterschiede in der Kinetik der Aktion des HDI sein f&uuml;r die klinische Anwendung dieser Verbindungen wichtig.

Charakterisierung der in-vitro-Aktivit&auml;t von der Depsipeptide Histon Deacetylase Inhibitor Spiruchostatin A.

Abbiamo recentemente completato la sintesi totale di spiruchostatin A, un prodotto naturale depsipeptide con somiglianze strutturali vicine a FK228, un inibitore di deacetylase (HDAC) dell&#39;istone (HDI) attualmente in corso di valutazione in studi clinici per il cancro. Qui riportiamo una caratterizzazione dettagliata dell&#39;attivit&agrave; in vitro di spiruchostatin A. Spiruchostatin A era un inibitore potente (sub-nM) di attivit&agrave; HDAC in vitro e ha agito come un profarmaco, che richiedono la riduzione di attivit&agrave; di classe I. A Spiruchostatin era un inibitore potente (bassa nM) la crescita di varie linee di cellule di cancro. Spiruchostatin A-induced acetilazione dei residui di lisina specifici all&#39;interno di istoni H3 e H4 e aumenta l&#39;espressione di p21(cip1/waf1), ma non ha indotto l&#39;acetilazione dell&#39;alfa-tubulina. Spiruchostatin A anche indotta dal ciclo delle cellule arresto e differenziazione delle cellule morte in cellule di cancro al seno MCF7. Come FK228, spiruchostatin A era un induttore e il substrato della pompa di efflusso del farmaco ABCB1. Considerando che la spiruchostatin A e acetilazione FK228-indotta dell&#39;istone prolungate, hydroxamate acetilazione dell&#39;istone di breve durata-HDI-indotta. Utilizzando un sottoinsieme dei geni HDI-target identificato da analisi di microarray, abbiamo dimostrato che queste differenze nella cinetica di acetilazione dell&#39;istone tra HDI correlata con differenze nella cinetica di induzione o repressione di geni bersaglio specifico. I nostri risultati dimostrano che spiruchostatin A &egrave; un potente inibitore della classe I HDACs e agente anti-cancro. Differenze nella cinetica di azione dell&#39;HDI possono essere importante per l&#39;applicazione clinica di questi composti.

Caratterizzazione dell&#39;attivit&agrave; in vitro del depsipeptide istone deacetylase dell&#39;inibitore spiruchostatin A.

&#x6211;&#x3005; &#x306F;&#x6700;&#x8FD1;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x30C7;&#x30D7;&#x30B7;&#x30DA;&#x30D7;&#x30C1;&#x30C9;&#x81EA;&#x7136;&#x88FD;&#x54C1; FK228&#x3001;&#x304C;&#x3093;&#x306E;&#x81E8;&#x5E8A;&#x8A66;&#x9A13;&#x306B;&#x304A;&#x3051;&#x308B;&#x73FE;&#x5728;&#x8A55;&#x4FA1;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x8131;&#x30A2;&#x30BB;&#x30C1;&#x30EB;&#x5316;&#x9175;&#x7D20; &#xFF08;HDAC&#xFF09; &#x963B;&#x5BB3; (HDI) &#x306B;&#x8FD1;&#x3044;&#x69CB;&#x9020;&#x985E;&#x4F3C;&#x6027;&#x3092;&#x6301;&#x3064;&#x306E;&#x5168;&#x5408;&#x6210;&#x3092;&#x5B8C;&#x4E86;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x8A73;&#x7D30;&#x306A;&#x7279;&#x6027;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A. &#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x3067;&#x3057;&#x305F;&#x5F37;&#x529B;&#x306A; (&#x30B5;&#x30D6; nM) &#x963B;&#x5BB3;&#x5264;&#x306E; in vitro &#x6D3B;&#x6027;&#x306E;&#x5831;&#x544A;&#x30AF;&#x30E9;&#x30B9; I HDAC &#x6D3B;&#x52D5; in vitro &#x304A;&#x3088;&#x3073;&#x524A;&#x6E1B;&#x3092;&#x5FC5;&#x8981;&#x3068;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x6D3B;&#x52D5;&#x306F;&#x30D7;&#x30ED;&#x30C9;&#x30E9;&#x30C3;&#x30B0;&#x3068;&#x3057;&#x3066;&#x884C;&#x52D5;&#x3057;&#x307E;&#x3059;&#x3002;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x306F;&#x5F37;&#x529B;&#x306A; (&#x4F4E; nM) &#x963B;&#x5BB3;&#x5264;&#x306E;&#x69D8;&#x3005; &#x306A;&#x764C;&#x7D30;&#x80DE;&#x306E;&#x6210;&#x9577;&#x3060;&#x3063;&#x305F;&#x3002;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x306B;&#x3088;&#x308B;&#x30A2;&#x30BB;&#x30C1;&#x30EB;&#x5316;&#x30D2;&#x30B9;&#x30C8;&#x30F3; H3&#x3001;H4&#x3001;&#x5185;&#x306E;&#x7279;&#x5B9A;&#x306E;&#x30EA;&#x30B8;&#x30F3;&#x6B8B;&#x57FA;&#x306E; p21(cip1/waf1) &#x306E;&#x767A;&#x73FE;&#x304C;&#x5897;&#x52A0;&#x3057;&#x305F;&#x304C;&#x3001;&alpha;-&#x30C1;&#x30E5;&#x30FC;&#x30D6;&#x30EA;&#x30F3;&#x306E;&#x30A2;&#x30BB;&#x30C1;&#x30EB;&#x5316;&#x8A98;&#x5C0E;&#x3055;&#x308C;&#x306A;&#x304B;&#x3063;&#x305F;&#x3002;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x306F; MCF7 &#x4E73;&#x764C;&#x7D30;&#x80DE;&#x306B;&#x3082;&#x7D30;&#x80DE;&#x5468;&#x671F;&#x306E;&#x902E;&#x6355;&#x3001;&#x5206;&#x5316;&#x3001;&#x7D30;&#x80DE;&#x6B7B;&#x3092;&#x8A98;&#x5C0E;&#x3057;&#x307E;&#x3059;&#x3002;FK228 &#x306E;&#x3088;&#x3046;&#x306A;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x306F;&#x4E21;&#x65B9;&#x3001;&#x30A4;&#x30F3;&#x30C7;&#x30E5;&#x30FC;&#x30B5;&#x3068;&#x57FA;&#x677F; ABCB1 &#x85AC;&#x5264;&#x6392;&#x51FA;&#x30DD;&#x30F3;&#x30D7;&#x306E;&#x3060;&#x3063;&#x305F;&#x3002;&#x4E00;&#x65B9;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x3068; FK228 &#x306B;&#x3088;&#x308B;&#x9577;&#x671F;&#x306E;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x306E;&#x30A2;&#x30BB;&#x30C1;&#x30EB;&#x5316;&#x3001;hydroxamate HDI &#x8A98;&#x8D77;&#x77ED;&#x547D;&#x306E;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x306E;&#x30A2;&#x30BB;&#x30C1;&#x30EB;&#x5316;&#x3002;HDI &#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x30DE;&#x30A4;&#x30AF;&#x30ED; &#x30A2;&#x30EC;&#x30A4;&#x5206;&#x6790;&#x306B;&#x3088;&#x3063;&#x3066;&#x7279;&#x5B9A;&#x306E;&#x30B5;&#x30D6;&#x30BB;&#x30C3;&#x30C8;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x3001;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x306E;&#x30A2;&#x30BB;&#x30C1;&#x30EB;&#x5316; HDI &#x306E;&#x9593;&#x306E;&#x901F;&#x5EA6;&#x306E;&#x9055;&#x3044;&#x304C;&#x8A98;&#x5C0E;&#x306E;&#x901F;&#x5EA6;&#x3084;&#x7279;&#x5B9A;&#x306E;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x6291;&#x5727;&#x306E;&#x9055;&#x3044;&#x3068;&#x76F8;&#x95A2;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005; &#x306E;&#x7D50;&#x679C;&#x306F;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; A &#x3092;&#x793A;&#x3059;&#x79C1;&#x306F;&#x3042;&#x308A;&#x3001;&#x6297;&#x60AA;&#x6027;&#x816B;&#x760D;&#x5264;&#x30AF;&#x30E9;&#x30B9;&#x306E;&#x5F37;&#x529B;&#x306A;&#x963B;&#x5BB3;&#x5264;&#x3067;&#x3059;&#x3002;HDI &#x306E;&#x4F5C;&#x7528;&#x306E;&#x901F;&#x5EA6;&#x8AD6;&#x306E;&#x9055;&#x3044;&#x306F;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x5316;&#x5408;&#x7269;&#x306E;&#x81E8;&#x5E8A;&#x5FDC;&#x7528;&#x306B;&#x91CD;&#x8981;&#x304B;&#x3082;&#x3057;&#x308C;&#x306A;&#x3044;&#x3002;

&#x30C7;&#x30D7;&#x30B7;&#x30DA;&#x30D7;&#x30C1;&#x30C9; &#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x8131;&#x30A2;&#x30BB;&#x30C1;&#x30EB;&#x5316;&#x9175;&#x7D20;&#x963B;&#x5BB3;&#x7269;&#x8CEA;&#x30B9;&#x30D4;&#x30EB;&#x30B3;&#x30B9;&#x30BF;&#x30C1;&#x30F3; a. &#x306E; in vitro &#x6D3B;&#x6027;&#x306E;&#x7279;&#x5FB4;

&#xC6B0;&#xB9AC;&#xB294; &#xCD5C;&#xADFC; spiruchostatin A, FK228, &#xD604;&#xC7AC; &#xC554;&#xC5D0; &#xB300; &#xD55C; &#xC784;&#xC0C1; &#xC2DC;&#xD5D8;&#xC5D0;&#xC11C; &#xD3C9;&#xAC00; &#xB418; &#xACE0; histone deacetylase (HDAC) &#xC5B5;&#xC81C;&#xC81C; (HDI)&#xC5D0; &#xAC00;&#xAE4C;&#xC6B4; &#xAD6C;&#xC870;&#xC801; &#xC720;&#xC0AC;&#xC131;&#xACFC; depsipeptide &#xC790;&#xC5F0; &#xC81C;&#xD488;&#xC758; &#xCD1D; &#xD569;&#xC131; &#xC644;&#xB8CC;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC; &#xBCF4;&#xACE0; spiruchostatin A. Spiruchostatin A&#xB294;&#xC758; &#xC801; &#xC815;&#xB7C9; (&#xD558;&#xC704; nM) &#xC5B5;&#xC81C;&#xC81C;&#xC758; &#xCCB4; &#xC678; &#xD65C;&#xB3D9;&#xC758; &#xC138;&#xBD80; &#xBB18;&#xC0AC;&#xAC00; &#xD074;&#xB798;&#xC2A4; I HDAC &#xD65C;&#xB3D9; &#xCCB4; &#xC678; &#xBC0F; &#xD65C;&#xB3D9;&#xC5D0; &#xB300; &#xD55C; &#xAC10;&#xC18C;&#xB97C; &#xD544;&#xC694;&#xB85C; prodrug&#xC73C;&#xB85C; &#xD589;&#xB3D9;. Spiruchostatin A&#xB294; &#xB2E4;&#xC591; &#xD55C; &#xC554; &#xC138;&#xD3EC; &#xB77C;&#xC778;&#xC758; &#xC131;&#xC7A5;&#xC758; &#xAC15;&#xB825;&#xD55C; (&#xB0AE;&#xC740; nM) &#xC5B5;&#xC81C;&#xC81C;. Spiruchostatin A &#xC720;&#xB3C4; acetylation &#xD788;&#xC2A4;&#xD1A4; H3 &#xBC0F; H4, &#xB0B4; &#xD2B9;&#xC815; lysine &#xC794;&#xB958;&#xBB3C;&#xC758; p21(cip1/waf1)&#xC758; &#xC2DD;&#xC744; &#xC99D;&#xAC00; &#xD558;&#xC9C0;&#xB9CC; &#xC54C;&#xD30C; tubulin acetylation &#xC720;&#xB3C4; &#xD558;&#xC9C0; &#xC54A;&#xC558;&#xB2E4;. Spiruchostatin A&#xB294; &#xB610;&#xD55C; MCF7 &#xC720;&#xBC29;&#xC554; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xC138;&#xD3EC; &#xC8FC;&#xAE30; &#xAC80;&#xAC70;, &#xBD84;&#xD654;&#xC640; &#xC138;&#xD3EC; &#xC8FD;&#xC74C;&#xC744; &#xC720;&#xB3C4;. FK228, &#xCC98;&#xB7FC; spiruchostatin A&#xB294; &#xC720;&#xB3C4; &#xBC0F; ABCB1 &#xB9C8;&#xC57D; &#xACBD;&#xACFC; &#xD38C;&#xD504;&#xC758; &#xAE30;&#xD310; &#xC774;&#xC5C8;&#xB2E4;. &#xBC18;&#xBA74; spiruchostatin A&#xC640; FK228 &#xC720;&#xB3C4; &#xC7A5;&#xAE30;&#xD654; &#xD788;&#xC2A4;&#xD1A4; acetylation, hydroxamate &#xB2E8;&#xAE30; &#xD788;&#xC2A4;&#xD1A4; HDI &#xC720;&#xB3C4; acetylation. HDI &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790; microarray &#xBD84;&#xC11D;&#xC5D0; &#xC758;&#xD574; &#xC2DD;&#xBCC4; &#xB41C; &#xD558;&#xC704; &#xC9D1;&#xD569;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC;, &#xC6B0;&#xB9AC;&#xB294; HDI &#xC0AC;&#xC774; histone acetylation&#xC758; &#xC18D;&#xB3C4; &#xB860;&#xC5D0;&#xC11C; &#xC774;&#xB7EC;&#xD55C; &#xCC28;&#xC774;&#xC758; &#xC720;&#xB3C4; &#xC18D;&#xB3C4; &#xB860; &#xB610;&#xB294; &#xD2B9;&#xC815; &#xB300;&#xC0C1; &#xC720;&#xC804;&#xC790;&#xC758; &#xC5B5;&#xC81C;&#xC758; &#xCC28;&#xC774;&#xC640; &#xC0C1;&#xAD00; &#xC2DC;&#xC5F0;. &#xC6B0;&#xB9AC;&#xC758; &#xACB0;&#xACFC; spiruchostatin A &#xC785;&#xC99D; &#xB0B4;&#xAC00; HDACs &#xBC0F; &#xC548;&#xD2F0;-&#xC554; &#xC5D0;&#xC774;&#xC804;&#xD2B8; &#xD074;&#xB798;&#xC2A4;&#xC758; &#xAC15;&#xB825;&#xD55C; &#xC5B5;&#xC81C;&#xC81C; &#xC774;&#xB2E4;. HDI&#xC758; &#xB3D9;&#xC791; &#xC18D;&#xB3C4;&#xC758; &#xCC28;&#xC774; &#xC774;&#xB7EC;&#xD55C; &#xD654;&#xD569;&#xBB3C;&#xC758; &#xC784;&#xC0C1; &#xC801;&#xC6A9;&#xC5D0; &#xB300; &#xD55C; &#xC911;&#xC694; &#xD55C; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;.

Depsipeptide &#xD788;&#xC2A4;&#xD1A4; deacetylase &#xC5B5;&#xC81C;&#xC81C; spiruchostatin 1. &#xCCB4; &#xC678; &#xD65C;&#xB3D9;&#xC758; &#xD2B9;&#xC131;

Recientemente hemos completado la s&iacute;ntesis total de spiruchostatin A, un producto natural de depsip&eacute;ptido con semejanzas estructurales cercanas a FK228, un inhibidor de la deacetilasa (HDAC) de la histona (HDI) actualmente est&aacute; evaluando en ensayos cl&iacute;nicos para el c&aacute;ncer. Presentamos una caracterizaci&oacute;n detallada de la actividad in vitro de spiruchostatin A. Spiruchostatin A es un inhibidor potente (sub-nM) de clase I actividad HDAC in vitro y actuaba como un prof&aacute;rmaco, que requieren reducci&oacute;n de la actividad. Spiruchostatin A es un inhibidor potente (baja nM) del crecimiento de varias l&iacute;neas celulares de c&aacute;ncer. Inducida por Spiruchostatin A la acetilaci&oacute;n de residuos de lisina espec&iacute;fica dentro de las histonas H3 y H4 y aument&oacute; la expresi&oacute;n de p21(cip1/waf1), pero no indujo la acetilaci&oacute;n de alfa-tubulina. Spiruchostatin A tambi&eacute;n indujo ciclo celular detenci&oacute;n, diferenciaci&oacute;n y muerte celular en c&eacute;lulas de c&aacute;ncer de mama MCF7. Como FK228, spiruchostatin A fue un inductor y sustrato de la bomba de eflujo ABCB1 drogas. Mientras que spiruchostatin A y la acetilaci&oacute;n de histonas prolongada inducida por FK228, hydroxamate la acetilaci&oacute;n de histonas corta vida inducida por el IDH. Utilizando un subconjunto de genes de HDI-Diana identificado por an&aacute;lisis de microarrays, demostramos que estas diferencias en la cin&eacute;tica de la acetilaci&oacute;n de histonas entre IDH correlacionaron con diferencias en la cin&eacute;tica de inducci&oacute;n o represi&oacute;n de genes espec&iacute;ficos. Nuestros resultados demuestran que spiruchostatin A es un inhibidor potente de la clase I HDAC y agente anticanceroso. Diferencias en la cin&eacute;tica de acci&oacute;n del IDH pueden ser importantes para la aplicaci&oacute;n cl&iacute;nica de estos compuestos.

Caracterizaci&oacute;n de la actividad in vitro de la depsip&eacute;ptido Histona deacetilasa inhibidor spiruchostatin A.

The folding of chromatin into topologically constrained loop domains is essential for genomic function. We have identified genomic anchors that define the organization of chromatin loop domains across the human major histocompatibility complex (MHC). This locus contains critical genes for immunity and is associated with more diseases than any other region of the genome. Classical MHC genes are expressed in a cell type-specific pattern and can be induced by cytokines such as interferon-gamma (IFNG). Transcriptional activation of the MHC was associated with a reconfiguration of chromatin architecture resulting from the formation of additional genomic anchors. These findings suggest that the dynamic arrangement of genomic anchors and loops plays a role in transcriptional regulation.

Reconfiguration of genomic anchors upon transcriptional activation of the human major histocompatibility complex.

To develop a model of X-linked inhibitor of apoptosis (XIAP) down regulation in colorectal cancer cell lines. This may be used to determine whether combination strategies have clinical potential.

Stable XIAP knockdown clones of HCT116 colon cancer cells are more sensitive to TRAIL, taxanes and irradiation in vitro.

The histone chaperone Vps75 forms a complex with, and stimulates the activity of, the histone acetyltransferase Rtt109. However, Vps75 can also be isolated on its own and might therefore possess Rtt109-independent functions. Analysis of epistatic miniarray profiles showed that VPS75 genetically interacts with factors involved in transcription regulation whereas RTT109 clusters with genes linked to DNA replication/repair. Additional genetic and biochemical experiments revealed a close relationship between Vps75 and RNA polymerase II. Furthermore, Vps75 is recruited to activated genes in an Rtt109-independent manner, and its genome-wide association with genes correlates with transcription rate. Expression microarray analysis identified a number of genes whose normal expression depends on VPS75. Interestingly, histone H2B dynamics at some of these genes are consistent with a role for Vps75 in histone H2A/H2B eviction/deposition during transcription. Indeed, reconstitution of nucleosome disassembly using the ATP-dependent chromatin remodeler Rsc and Vps75 revealed that these proteins can cooperate to remove H2A/H2B dimers from nucleosomes. These results indicate a role for Vps75 in nucleosome dynamics during transcription, and importantly, this function appears to be largely independent of Rtt109.

An rtt109-independent role for vps75 in transcription-associated nucleosome dynamics.

Understanding how the immune system in patients with cancer interacts with malignant cells is critical for the development of successful immunotherapeutic strategies. We studied peripheral blood from newly diagnosed patients with acute myeloid leukemia (AML) to assess the impact of this disease on the patients' T cells. The absolute number of peripheral blood T cells is increased in AML compared with healthy controls. An increase in the absolute number of CD3+56+ cells was also noted. Gene expression profiling on T cells from AML patients compared with healthy donors demonstrated global differences in transcription suggesting aberrant T-cell activation patterns. These gene expression changes differ from those observed in chronic lymphocytic leukemia (CLL), indicating the heterogeneous means by which different tumors evade the host immune response. However, in common with CLL, differentially regulated genes involved in actin cytoskeletal formation were identified, and therefore the ability of T cells from AML patients to form immunologic synapses was assessed. Although AML T cells could form conjugates with autologous blasts, their ability to form immune synapses and recruit phosphotyrosine signaling molecules to the synapse was significantly impaired. These findings identify T-cell dysfunction in AML that may contribute to the failure of a host immune response against leukemic blasts.

Peripheral blood T cells in acute myeloid leukemia (AML) patients at diagnosis have abnormal phenotype and genotype and form defective immune synapses with AML blasts.

Cells generate new organelles when stimulated by extracellular factors to grow and divide; however, little is known about how growth and mitogenic signalling pathways regulate organelle biogenesis. Using mitochondria as a model organelle, we have investigated this problem in primary Schwann cells, for which distinct factors act solely as mitogens (neuregulin) or as promoters of cell growth (insulin-like growth factor 1; IGF1). We find that neuregulin and IGF1 act synergistically to increase mitochondrial biogenesis and mitochondrial DNA replication, resulting in increased mitochondrial density in these cells. Moreover, constitutive oncogenic Ras signalling results in a further increase in mitochondrial density. This synergistic effect is seen at the global transcriptional level, requires both the ERK and phosphoinositide 3-kinase (PI3K) signalling pathways and is mediated by the transcription factor ERRalpha. Interestingly, the effect is independent of Akt-TOR signalling, a major regulator of cell growth in these cells. This separation of the pathways that drive mitochondrial biogenesis and cell growth provides a mechanism for the modulation of mitochondrial density according to the metabolic requirements of the cell.

Extracellular growth factors and mitogens cooperate to drive mitochondrial biogenesis.

mRNA &#x672C;&#x5730;&#x5316;&#x662F;&#x4E00;&#x4E2A;&#x8FDB;&#x5316;&#x7684;&#x5B88;&#x6052;&#x7684;&#x673A;&#x5236;&#x7684;&#x57FA;&#x7840;&#x5EFA;&#x7ACB;&#x7EC6;&#x80DE;&#x6781;&#x6027;&#x548C;&#x4E13;&#x95E8;&#x7684;&#x7EC6;&#x80DE;&#x529F;&#x80FD;&#x3002;&#x82E5;&#x8981;&#x6807;&#x8BC6;&#x672C;&#x5730;&#x5316;&#x53D1;&#x5C55;&#x795E;&#x7ECF;&#x5143;&#x4E9A;&#x7EC6;&#x80DE;&#x8F66;&#x53A2;&#x5185;&#x7684;&#x5206;&#x6CCC;&#xFF0C;&#x6211;&#x4EEC;&#x91C7;&#x53D6;&#x539F;&#x59CB;&#x7684;&#x4EA4;&#x611F;&#x795E;&#x7ECF;&#x5143;&#x7684;&#x6761;&#x5757;&#x5206;&#x5272;&#x7684;&#x6587;&#x5316;&#x4E0E;&#x5E8F;&#x8D2F;&#x5206;&#x6790;&#x57FA;&#x56E0;&#x8868;&#x8FBE; (SAGE) &#x76F8;&#x7ED3;&#x5408;&#x3002;&#x610F;&#x5916;&#x7684;&#x662F;&#xFF0C;&#x795E;&#x7ECF;&#x8F74;&#x7A81;&#x4E2D;&#x6700;&#x4E30;&#x5BCC;&#x7684;&#x6210;&#x7EE9;&#x5355;&#x662F; mRNA &#x808C;&#x9187;&#x5076;&#x8054;-1 (Impa1)&#xFF0C;&#x89C4;&#x5B9A;&#x4E86;&#x808C;&#x9187;&#x5468;&#x671F;&#x548C;&#x795E;&#x7ECF;&#x5143;&#x4E2D;&#x9502;&#x7684;&#x4E3B;&#x8981;&#x76EE;&#x6807;&#x7684;&#x5173;&#x952E;&#x9176;&#x3002;&#x5185; Impa1 mRNA 3&#39; &#x975E;&#x7FFB;&#x8BD1;&#x533A;&#x7684;&#x65B0;&#x5B9A;&#x4F4D;&#x5143;&#x7D20;&#x7684;&#x5177;&#x4F53;&#x76EE;&#x6807; Impa1 &#x8BB0;&#x5F55;&#xFF0C;&#x4F7F;&#x4EA4;&#x611F;&#x795E;&#x7ECF;&#x795E;&#x7ECF;&#x5143;&#x8F74;&#x7A81;&#x548C;&#x795E;&#x7ECF;&#x751F;&#x957F;&#x56E0;&#x5B50; (NGF) &#x56DE;&#x5E94;&#x53D7;&#x89C4;&#x7BA1;&#x672C;&#x5730; IMPA1 &#x7FFB;&#x8BD1;&#x3002;&#x9009;&#x62E9;&#x6027;&#x6C89;&#x9ED8;&#x7684; IMPA1 &#x5408;&#x6210;&#x7684;&#x795E;&#x7ECF;&#x8F74;&#x7A81;&#x51CF;&#x5C11;&#x6838; CREB &#x6D3B;&#x5316;&#x548C;&#x81F4;&#x5F25;&#x6F2B;&#x6027;&#x8F74;&#x7D22;&#x53D8;&#x6027;&#x3002;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x63D0;&#x4F9B;&#x6D1E;&#x5BDF; mRNA &#x5728;&#x795E;&#x7ECF;&#x8F74;&#x7A81;&#x8FD0;&#x8F93;&#x548C;&#x63ED;&#x793A;&#x65B0;&#x7684;&#x795E;&#x7ECF;&#x751F;&#x957F;&#x56E0;&#x5B50;&#x54CD;&#x5E94;&#x672C;&#x5730;&#x5316;&#x5143;&#x7D20;&#x6307;&#x793A;&#x76EE;&#x6807;&#x548C;&#x672C;&#x5730;&#x7FFB;&#x8BD1;&#x7684;&#x8F74;&#x7A81;&#x7684;&#x8C08;&#x8BDD;&#x5185;&#x5BB9;&#x3002;

&#x795E;&#x7ECF;&#x751F;&#x957F;&#x56E0;&#x5B50;&#x53CD;&#x5E94;&#x7684;&#x5143;&#x7D20;&#x7684;&#x76EE;&#x6807;&#x5230;&#x540C;&#x60C5;&#x795E;&#x7ECF;&#x5143;&#x8F74;&#x7A81;&#x808C;&#x9187;&#x5076;&#x8054;-1 mRNA&#x3002;

mRNA localization is an evolutionary conserved mechanism that underlies the establishment of cellular polarity and specialized cell functions. To identify mRNAs localized in subcellular compartments of developing neurons, we took an original approach that combines compartmentalized cultures of rat sympathetic neurons and sequential analysis of gene expression (SAGE). Unexpectedly, the most abundant transcript in axons was mRNA for myo-inositol monophosphatase-1 (Impa1), a key enzyme that regulates the inositol cycle and the main target of lithium in neurons. A novel localization element within the 3' untranslated region of Impa1 mRNA specifically targeted Impa1 transcript to sympathetic neuron axons and regulated local IMPA1 translation in response to nerve growth factor (NGF). Selective silencing of IMPA1 synthesis in axons decreased nuclear CREB activation and induced axonal degeneration. These results provide insights into mRNA transport in axons and reveal a new NGF-responsive localization element that directs the targeting and local translation of an axonal transcript.

An NGF-responsive element targets myo-inositol monophosphatase-1 mRNA to sympathetic neuron axons.

localisation des ARNm est un m&#xE9;canisme &#xE9;volutif conserv&#xE9; qui sous-tend l&#x26;#39;&#xE9;tablissement de la polarit&#xE9; cellulaire et les fonctions des cellules sp&#xE9;cialis&#xE9;es. Pour identifier les ARNm localis&#xE9;s dans des compartiments subcellulaires de neurones en d&#xE9;veloppement, nous avons pris une approche originale qui combine les cultures compartiment&#xE9;es de rat neurones sympathiques et de l&#x26;#39;analyse s&#xE9;quentielle de l&#x26;#39;expression g&#xE9;nique (SAGE). De fa&#xE7;on inattendue, la transcription la plus abondante dans les axones &#xE9;tait ARNm de myo-inositol monophosphatase-1 (Impa1), une enzyme cl&#xE9; qui r&#xE9;gule le cycle de l&#x26;#39;inositol et la principale cible de lithium dans les neurones. Un &#xE9;l&#xE9;ment de localisation roman dans la r&#xE9;gion 3 &#x26;#39;non traduite de l&#x26;#39;ARNm cibl&#xE9; Impa1 Impa1 transcription axones des neurones sympathiques et r&#xE9;glement&#xE9; locale IMPA1 traduction en r&#xE9;ponse au facteur de croissance nerveuse (NGF). Silence s&#xE9;lectif de la synth&#xE8;se dans les axones IMPA1 diminu&#xE9; nucl&#xE9;aire activation induite par la prot&#xE9;ine CREB et une d&#xE9;g&#xE9;n&#xE9;rescence axonale. Ces r&#xE9;sultats donnent un aper&#xE7;u de transport d&#x26;#39;ARNm dans les axones et de r&#xE9;v&#xE9;ler un &#xE9;l&#xE9;ment de localisation nouvelle NGF-sensible qui dirige la traduction de ciblage et de locaux d&#x26;#39;une transcription axonale.

Un &#xE9;l&#xE9;ment sensible NGF Cibles myo-inositol-1 monophosphatase ARNm axones des neurones sympathiques

mRNA Lokalisierung ist ein evolution&auml;r konservierte Mechanismus, der Errichtung einer zellul&auml;ren Polarit&auml;t und spezialisierte Zelle Funktionen zugrunde liegt. Ermittlung der mRNAs lokalisiert in subzellul&auml;ren F&auml;cher Neuronen zu entwickeln, haben wir einen originellen Ansatz, der Abteilen Kulturen der Ratte sympathischen Neuronen und sequentielle Analyse der Genexpression (SAGE) kombiniert. Unerwartet war die am h&auml;ufigsten vorkommenden Abschrift in Axonen mRNA f&uuml;r Myo-Inosit Monophosphatase-1 (Impa1), ein Schl&uuml;sselenzym, das der Inosit-Zyklus und das Hauptziel der Lithium in Neuronen reguliert. Eine neuartige Lokalisierung-Element innerhalb der 3&#39; un&uuml;bersetzte Region Impa1 mRNA gezielt speziell Impa1 Abschrift sympathischen Neuronen Axone und regulierten lokalen IMPA1 &Uuml;bersetzung als Reaktion auf Nerv Wachstumfaktor (NGF). Selektive Unterdr&uuml;ckender IMPA1 Synthese in Axonen verringerte sich um nukleare CREB-Aktivierung und induzierte axonale Degeneration. Diese Ergebnisse vermitteln Einblicke in mRNA-Transport in Axonen und offenbaren ein neues NGF-responsive Lokalisierung-Element, das die Ausrichtung und lokale &Uuml;bersetzung eine axonale Transkript leitet.

Ein NGF-responsive Element richtet sich an Myo-Inosit Monophosphatase-1 mRNA zu sympathischen Neuronen Axone.

localizzazione di mRNA &egrave; un meccanismo evolutivo conservato che sottende l&#39;istituzione della polarit&agrave; cellulare e le funzioni delle cellule specializzate. Per identificare i mRNAs localizzato in compartimenti subcellulari dei neuroni in via di sviluppo, abbiamo preso un approccio originale che unisce culture compartimenti stagni dei neuroni simpatici rat e sequenziale analisi di espressione genica (SAGE). Inaspettatamente, la trascrizione pi&ugrave; abbondante in assoni era mRNA per myo-inositolo monophosphatase-1 (Impa1), un enzima chiave che regola il ciclo di inositolo e il principale bersaglio di litio in neuroni. Un elemento di localizzazione romanzo all&#39;interno della regione non tradotta 3&#39; del mRNA di Impa1 mirati specificamente Impa1 trascrizione al simpatico neurone assoni e regolamentati locali IMPA1 traduzione in risposta a nerve growth factor (NGF). Silenziamento selettivo della sintesi di IMPA1 in assoni diminuita attivazione CREB nucleare e indotta degenerazione assonale. Questi risultati forniscono informazioni sul trasporto di mRNA in assoni e rivelano un nuovo elemento di localizzazione NGF-reattivo che dirige la traduzione di targeting e locale di una trascrizione assonale.

Un elemento reattivo NGF obiettivi myo-inositolo monophosphatase-1 mRNA di assoni neurone simpatico.

mRNA&#x306E;&#x5C40;&#x5728;&#x306F;&#x3001;&#x7D30;&#x80DE;&#x6975;&#x6027;&#x3068;&#x7279;&#x6B8A;&#x306A;&#x7D30;&#x80DE;&#x6A5F;&#x80FD;&#x306E;&#x78BA;&#x7ACB;&#x306E;&#x57FA;&#x790E;&#x3068;&#x306A;&#x308B;&#x9032;&#x5316;&#x306E;&#x4FDD;&#x5B58;&#x3055;&#x308C;&#x305F;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3067;&#x3059;&#x3002;&#x958B;&#x767A;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x7D30;&#x80DE;&#x5185;&#x30B3;&#x30F3;&#x30D1;&#x30FC;&#x30C8;&#x30E1;&#x30F3;&#x30C8;&#x306B;&#x5C40;&#x5728;&#x3059;&#x308B;mRNA&#x3092;&#x540C;&#x5B9A;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3001;&#x6211;&#x3005;&#x306F;&#x3001;&#x30E9;&#x30C3;&#x30C8;&#x306E;&#x4EA4;&#x611F;&#x795E;&#x7D4C;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x3068;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x306E;&#x9023;&#x7D9A;&#x5206;&#x6790;&#xFF08;SAGE&#xFF09;&#x306E;&#x533A;&#x5206;&#x306E;&#x6587;&#x5316;&#x3092;&#x7D44;&#x307F;&#x5408;&#x308F;&#x305B;&#x305F;&#x72EC;&#x5275;&#x7684;&#x306A;&#x30A2;&#x30D7;&#x30ED;&#x30FC;&#x30C1;&#x3092;&#x53D6;&#x3063;&#x305F;&#x3002;&#x4E88;&#x671F;&#x305B;&#x305A;&#x3001;&#x8EF8;&#x7D22;&#x306E;&#x4E2D;&#x3067;&#x6700;&#x3082;&#x8C4A;&#x5BCC;&#x306A;&#x8EE2;&#x5199;&#x7523;&#x7269;&#x306F;&#x3001;&#x30DF;&#x30AA;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;monophosphatase-1&#xFF08;Impa1&#xFF09;&#x3001;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;&#x30B5;&#x30A4;&#x30AF;&#x30EB;&#x3068;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306B;&#x304A;&#x3051;&#x308B;&#x30EA;&#x30C1;&#x30A6;&#x30E0;&#x306E;&#x30E1;&#x30A4;&#x30F3;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x9375;&#x3068;&#x306A;&#x308B;&#x9175;&#x7D20;&#x306E;mRNA&#x3067;&#x3042;&#x3063;&#x305F;&#x3002; Impa1 mRNA&#x306E;3 &#x26;#39;&#x975E;&#x7FFB;&#x8A33;&#x9818;&#x57DF;&#x5185;&#x306E;&#x5C0F;&#x8AAC;&#x306E;&#x30ED;&#x30FC;&#x30AB;&#x30E9;&#x30A4;&#x30BA;&#x8981;&#x7D20;&#x306F;&#x3001;&#x7279;&#x306B;&#x795E;&#x7D4C;&#x6210;&#x9577;&#x56E0;&#x5B50;&#xFF08;NGF&#xFF09;&#x306B;&#x5FDC;&#x7B54;&#x3057;&#x3066;&#x3001;&#x4EA4;&#x611F;&#x795E;&#x7D4C;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x8EF8;&#x7D22;&#x3068;&#x898F;&#x5236;&#x30ED;&#x30FC;&#x30AB;&#x30EB;IMPA1&#x7FFB;&#x8A33;&#x306B;Impa1&#x8EE2;&#x5199;&#x7523;&#x7269;&#x3092;&#x5BFE;&#x8C61;&#x3068;&#x3057;&#x305F;&#x3002;&#x8EF8;&#x7D22;&#x306E;IMPA1&#x5408;&#x6210;&#x306E;&#x9078;&#x629E;&#x7684;&#x30B5;&#x30A4;&#x30EC;&#x30F3;&#x30B7;&#x30F3;&#x30B0;&#x306F;&#x3001;&#x6838;&#x306E;CREB&#x306E;&#x6D3B;&#x6027;&#x5316;&#x200B;&#x200B;&#x3068;&#x8A98;&#x5C0E;&#x8EF8;&#x7D22;&#x5909;&#x6027;&#x3092;&#x6E1B;&#x5C11;&#x3055;&#x305B;&#x305F;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D50;&#x679C;&#x306F;&#x3001;&#x8EF8;&#x7D22;&#x306E;mRNA&#x306E;&#x8F38;&#x9001;&#x3078;&#x306E;&#x6D1E;&#x5BDF;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x3001;&#x8EF8;&#x7D22;&#x306E;&#x8EE2;&#x5199;&#x306E;&#x6A19;&#x7684;&#x3068;&#x5C40;&#x6240;&#x7684;&#x7FFB;&#x8A33;&#x3092;&#x6307;&#x793A;&#x3059;&#x308B;&#x65B0;&#x3057;&#x3044;NGF&#x5FDC;&#x7B54;&#x30ED;&#x30FC;&#x30AB;&#x30E9;&#x30A4;&#x30BA;&#x8981;&#x7D20;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;&#x3002;

NGF&#x5FDC;&#x7B54;&#x30A8;&#x30EC;&#x30E1;&#x30F3;&#x30C8;&#x306F;&#x4EA4;&#x611F;&#x795E;&#x7D4C;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x306E;&#x8EF8;&#x7D22;&#x306B;&#x30DF;&#x30AA;&#x30A4;&#x30CE;&#x30B7;&#x30C8;&#x30FC;&#x30EB;Monophosphatase-1 mRNA&#x3092;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;

mRNA &#xC9C0;&#xC5ED;&#xD654; &#xC140;&#xB8F0;&#xB7EC; &#xADF9;&#xC131; &#xBC0F; &#xD2B9;&#xC218; &#xD55C; &#xC140; &#xD568;&#xC218; &#xC124;&#xB9BD;&#xC758; &#xAE30;&#xBC18;&#xC774; &#xB418;&#xB294; &#xC9C4;&#xD654; &#xBCF4;&#xC874;&#xB41C; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC785;&#xB2C8;&#xB2E4;. &#xB274;&#xB7F0; &#xAC1C;&#xBC1C;&#xC758; subcellular &#xAD6C;&#xD68D;&#xC5D0; &#xC9C0;&#xC5ED;&#xD654; mRNAs &#xC2DD;&#xBCC4; &#xC950; &#xAD50;&#xAC10; &#xC2E0;&#xACBD;&#xC758; &#xBD84;&#xB958; &#xBB38;&#xD654;&#xC640; &#xC720;&#xC804;&#xC790; &#xBC1C;&#xD604; (&#xC138;&#xC774; &#xC9C0;)&#xC758; &#xC21C;&#xCC28; &#xBD84;&#xC11D;&#xC744; &#xACB0;&#xD569; &#xD558; &#xC5EC; &#xC6D0;&#xB798;&#xC758; &#xC811;&#xADFC; &#xBC29;&#xC2DD;&#xC744; &#xD588;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC608;&#xAE30;&#xCE58; &#xC54A;&#xAC8C;, axons&#xC5D0;&#xC11C; &#xAC00;&#xC7A5; &#xD48D;&#xBD80;&#xD55C; &#xC0AC;&#xBCF8;&#xC740; myo-&#xC774;&#xB178; &#xC2DC; &#xD1A8; monophosphatase-1 (Impa1) &#xC870;&#xC808; inositol &#xC0AC;&#xC774;&#xD074;&#xACFC; &#xB274;&#xB7F0;&#xC5D0;&#xC11C; &#xB9AC;&#xD2AC;&#xC758; &#xC8FC;&#xC694; &#xB300;&#xC0C1; &#xC8FC;&#xC694; &#xD6A8;&#xC18C; Mrna&#xB97C; &#xD588;&#xB2E4;. Impa1 Mrna&#xC758; 3&#39; &#xB9DE;&#xC774; &#xC601;&#xC5ED; &#xB0B4;&#xC5D0;&#xC11C; &#xC0C8;&#xB85C;&#xC6B4; &#xC9C0;&#xC5ED;&#xD654; &#xC694;&#xC18C; &#xD2B9;&#xD788; &#xAD50;&#xAC10; &#xC2E0;&#xACBD; &#xCD95; &#xC0AD;&#xACFC; &#xC2E0;&#xACBD; &#xC131;&#xC7A5; &#xC778;&#xC790; (NGF)&#xC5D0; &#xB300; &#xD55C; &#xC751;&#xB2F5; &#xB808; &#xADE4; &#xB808;&#xC774;&#xD2B8; &#xB41C; &#xB85C;&#xCEEC; IMPA1 &#xBC88;&#xC5ED;&#xC744; Impa1 &#xB300; &#xBCF8;&#xC744; &#xB300;&#xC0C1;. IMPA1 &#xD569;&#xC131; &#xCD95; &#xC0AD;&#xC758; &#xC120;&#xD0DD;&#xC801; &#xC785;&#xC744; &#xD575; CREB &#xD65C;&#xC131;&#xD654;&#xB97C; &#xAC10;&#xC18C; &#xD558; &#xACE0; axonal &#xBCC0;&#xC131; &#xC720;&#xBC1C;. &#xC774;&#xB7EC;&#xD55C; &#xACB0;&#xACFC; &#xCD95; &#xC0AD;&#xC758; mRNA &#xAD50;&#xD1B5;&#xC5D0; &#xB300; &#xD55C; &#xD1B5;&#xCC30;&#xB825;&#xC744; &#xC81C;&#xACF5; &#xD558; &#xACE0; axonal &#xC131;&#xC801; &#xB300;&#xC0C1; &#xBC0F; &#xB85C;&#xCEEC; &#xBC88;&#xC5ED;&#xC744; &#xC9C0;&#xD718;&#xD558; &#xACE0; &#xC0C8;&#xB85C;&#xC6B4; NGF &#xC751;&#xB2F5; &#xC9C0;&#xC5ED;&#xD654; &#xC694;&#xC18C;&#xB97C; &#xACF5;&#xAC1C;.

NGF &#xBC18;&#xC751; &#xC694;&#xC18C; myo-&#xC774;&#xB178; &#xC2DC; &#xD1A8; monophosphatase-1 &#xAD50;&#xAC10; &#xC2E0;&#xACBD; &#xCD95; &#xC0AD;&#xC744; Mrna&#xB97C; &#xB300;&#xC0C1;.

la localizaci&#xF3;n del ARNm, es un mecanismo evolutivo conservado que subyace a la creaci&#xF3;n de la polaridad celular y las funciones celulares especializadas. Para identificar los ARNm localizadas en compartimentos subcelulares de las neuronas en desarrollo, tomamos un enfoque original que combina las culturas en compartimientos de las ratas las neuronas simp&#xE1;ticas y an&#xE1;lisis secuencial de expresi&#xF3;n g&#xE9;nica (SAGE). Inesperadamente, la transcripci&#xF3;n m&#xE1;s abundante en los axones era ARNm de myo-inositol monofosfatasa-1 (Impa1), una enzima clave que regula el ciclo del inositol y el objetivo principal de litio en neuronas. Un elemento de localizaci&#xF3;n de la novela dentro de la regi&#xF3;n 3 &#x26;#39;no traducida del ARNm Impa1 espec&#xED;ficamente Impa1 transcripci&#xF3;n a axones de las neuronas simp&#xE1;ticas y regulado de traducci&#xF3;n local de IMPA1 en respuesta al factor de crecimiento nervioso (NGF). El silenciamiento selectivo de la s&#xED;ntesis de IMPA1 en los axones disminuy&#xF3; la activaci&#xF3;n de CREB nuclear y la degeneraci&#xF3;n axonal inducida. Estos resultados permiten conocer el transporte de ARNm en los axones y revelan un nuevo NGF elemento de respuesta de localizaci&#xF3;n que dirige la traducci&#xF3;n local de focalizaci&#xF3;n y una transcripci&#xF3;n axonal.

Un elemento que responde a NGF Objetivos mio-inositol-1 mRNA monofosfatasa de axones de las neuronas simp&#xE1;ticas

Proinflammatory cytokines are frequently observed in the tumor microenvironment, and chronic inflammation is involved in cancer initiation and progression. We show that cytokine signaling through the receptor subunit GP130-IL6ST and the kinase JAK1 generates actomyosin contractility through Rho-kinase dependent signaling. This pathway generates contractile force in stromal fibroblasts to remodel the extracellular matrix to create tracks for collective migration of squamous carcinoma cells and provides the high levels of actomyosin contractility required for migration of individual melanoma cells in the rounded, "amoeboid" mode. Thus, cytokine signaling can generate actomyosin contractility in both stroma and tumor cells. Strikingly, actomyosin contractility itself positively modulates activity of the transcription factor STAT3 downstream of JAK1, demonstrating positive feedback within the signaling network.

ROCK and JAK1 signaling cooperate to control actomyosin contractility in tumor cells and stroma.

We identified XAB1 in a proteomic screen for factors that interact with human RNA polymerase II (RNAPII). Because XAB1 has a conserved Saccharomyces cerevisiae homologue called Npa3, yeast genetics and biochemical analysis were used to dissect the significance of the interaction. Degron-dependent Npa3 depletion resulted in genome-wide transcription decreases, correlating with a loss of RNAPII from genes as measured by chromatin immunoprecipitation. Surprisingly, however, transcription in vitro was unaffected by Npa3, suggesting that it affects a process that is not required for transcription in yeast extracts. Indeed, Npa3 depletion in vivo affects nuclear localization of RNAPII; the polymerase accumulates in the cytoplasm. Npa3 is a member of the GPN-LOOP family of GTPases. Npa3 mutants that either cannot bind GTP or that bind but cannot hydrolyze it are inviable and unable to support nuclear transport of RNAPII. Surprisingly, we were unable to detect interactions between Npa3 and proteins in the classical importin α/β pathway for nuclear import. Interestingly, Npa3-RNAPII binding is significantly increased by the addition of GTP or its slowly hydrolyzable analogue guanosine 5'-3-O-(thio)triphosphate (GTPγS). Moreover, the Npa3 mutant that binds GTP, but cannot hydrolyze it, binds RNAPII even in the absence of added GTP, whereas the mutant that cannot bind GTP is unable to bind the polymerase. Together, our data suggest that Npa3 defines an unconventional pathway for nuclear import of RNAPII, which involves GTP-dependent binding of Npa3 to the polymerase.

GTP-dependent binding and nuclear transport of RNA polymerase II by Npa3 protein.

Transport of cargoes by kinesin-1 is essential for many cellular processes. Nevertheless, the number of proteins known to recruit kinesin-1 via its cargo binding light chain (KLC) is still quite small. We also know relatively little about the molecular features that define kinesin-1 binding. We now show that a bipartite tryptophan-based kinesin-1 binding motif, originally identified in Calsyntenin is present in A36, a vaccinia integral membrane protein. This bipartite motif in A36 is required for kinesin-1-dependent transport of the virus to the cell periphery. Bioinformatic analysis reveals that related bipartite tryptophan-based motifs are present in over 450 human proteins. Using vaccinia as a surrogate cargo, we show that regions of proteins containing this motif can function to recruit KLC and promote virus transport in the absence of A36. These proteins interact with the kinesin light chain outside the context of infection and have distinct preferences for KLC1 and KLC2. Our observations demonstrate that KLC binding can be conferred by a common set of features that are found in a wide range of proteins associated with diverse cellular functions and human diseases.

A kinesin-1 binding motif in vaccinia virus that is widespread throughout the human genome.

The radical response of peripheral nerves to injury (Wallerian degeneration) is the cornerstone of nerve repair. We show that activation of the transcription factor c-Jun in Schwann cells is a global regulator of Wallerian degeneration. c-Jun governs major aspects of the injury response, determines the expression of trophic factors, adhesion molecules, the formation of regeneration tracks and myelin clearance and controls the distinctive regenerative potential of peripheral nerves. A key function of c-Jun is the activation of a repair program in Schwann cells and the creation of a cell specialized to support regeneration. We show that absence of c-Jun results in the formation of a dysfunctional repair cell, striking failure of functional recovery, and neuronal death. We conclude that a single glial transcription factor is essential for restoration of damaged nerves, acting to control the transdifferentiation of myelin and Remak Schwann cells to dedicated repair cells in damaged tissue.

c-Jun reprograms Schwann cells of injured nerves to generate a repair cell essential for regeneration.

Angiogenesis is essential for solid tumour growth, whilst the molecular profiles of tumour blood vessels have been reported to be different between cancer types. Although presently available anti-angiogenic strategies are providing some promise for the treatment of some cancers it is perhaps not surprisingly that, none of the anti-angiogenic agents available work on all tumours. Thus, the discovery of novel anti-angiogenic targets, relevant to individual cancer types, is required. Using Affymetrix microarray analysis of laser-captured, CD31-positive blood vessels we have identified 63 genes that are upregulated significantly (5-72 fold) in angiogenic blood vessels associated with human invasive ductal carcinoma (IDC) of the breast as compared with blood vessels in normal human breast. We tested the angiogenic capacity of a subset of these genes. Genes were selected based on either their known cellular functions, their enriched expression in endothelial cells and/or their sensitivity to anti-VEGF treatment; all features implicating their involvement in angiogenesis. For example, RRM2, a ribonucleotide reductase involved in DNA synthesis, was upregulated 32-fold in IDC-associated blood vessels; ATF1, a nuclear activating transcription factor involved in cellular growth and survival was upregulated 23-fold in IDC-associated blood vessels and HEX-B, a hexosaminidase involved in the breakdown of GM2 gangliosides, was upregulated 8-fold in IDC-associated blood vessels. Furthermore, in silico analysis confirmed that AFT1 and HEX-B also were enriched in endothelial cells when compared with non-endothelial cells. None of these genes have been reported previously to be involved in neovascularisation. However, our data establish that siRNA depletion of Rrm2, Atf1 or Hex-B had significant anti-angiogenic effects in VEGF-stimulated ex vivo mouse aortic ring assays. Overall, our results provide proof-of-principle that our approach can identify a cohort of potentially novel anti-angiogenic targets that are likley to be, but not exclusivley, relevant to breast cancer.

Gene expression analysis in human breast cancer associated blood vessels.

&#x7EC6;&#x80DE;&#x6838;&#x7CD6;&#x4F53;&#x86CB;&#x767D; L29 (RPL29) &#x88AB;&#x8BA4;&#x4E3A;&#x662F;&#x91CD;&#x8981;&#x7684;&#x86CB;&#x767D;&#x8D28;&#x5408;&#x6210;&#xFF0C;&#x4F46;&#x4E4B;&#x524D;&#x4ECE;&#x672A;&#x89C1;&#x5176;&#x5728;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x8FC7;&#x7A0B;&#x7684;&#x529F;&#x80FD;&#x3002;&#x6211;&#x4EEC;&#x5DF2;&#x7ECF;&#x8868;&#x660E;&#x4EE5;&#x524D;&#x7F3A;&#x4E4F; &beta;3 &#x6574;&#x5408;&#x7D20;&#x5C0F;&#x9F20;&#x652F;&#x6301;&#x589E;&#x5F3A;&#x7684;&#x80BF;&#x7624;&#x8840;&#x7BA1;&#x751F;&#x6210;&#xFF0C;&#x56E0;&#x6B64;&#xFF0C;&#x5220;&#x9664;&#x7684;&#x5185;&#x76AE;&#x7EC6;&#x80DE; &alpha;v&beta;3 &#x53EF;&#x4EE5;&#x63D0;&#x4F9B;&#x4E00;&#x79CD;&#x65B9;&#x6CD5;&#x7684;&#x80BF;&#x7624;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x7684;&#x65B0;&#x578B;&#x76D1;&#x7BA1;&#x673A;&#x6784;&#x53D1;&#x73B0;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x5C06;&#x4ECB;&#x7ECD; RPL29 &beta;3 &#x7A7A;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x7684;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x548C;&#x86CB;&#x767D;&#x8D28;&#x4E00;&#x7EA7;&#x8868;&#x8FBE;&#x660E;&#x663E;&#x4E0A;&#x8C03;&#x3002;&#x6269;&#x589E;&#xFF0C;&#x6211;&#x4EEC;&#x663E;&#x793A; VEGF &#x523A;&#x6FC0;&#x5FAE;&#x8840;&#x7BA1;&#x53D1;&#x82BD;&#x663E;&#x8457;&#x51CF;&#x5C11; Rpl29 &#x6742;&#x5408;&#x6027;&#x548C; Rpl29 null &#x4E3B;&#x52A8;&#x8109;&#x73AF;&#x68C0;&#x6D4B;&#x76F8;&#x6BD4;&#x91CE;&#x751F;&#x7C7B;&#x578B;&#x7684;&#x63A7;&#x4EF6;&#x4E2D;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6211;&#x4EEC;&#x8FD8;&#x63D0;&#x4F9B; RPL29 &#x53EF;&#x8C03;&#x8282;&#x80BF;&#x7624;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x7684;&#x4F53;&#x5185;&#x8BC1;&#x636E;&#x3002;&#x80BF;&#x7624;&#x8840;&#x7BA1;&#x5BC6;&#x5EA6;&#x7684;&#x76AE;&#x4E0B;&#x79CD;&#x690D;&#x7684; Lewis &#x80BA;&#x764C; Rpl29 &#x7A81;&#x53D8;&#x5C0F;&#x9F20;&#x663E;&#x8457;&#x51CF;&#x5C11;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6D88;&#x8017;&#x7684; Rpl29 &#x5229;&#x7528; RNA &#x5E72;&#x6270;&#x6280;&#x672F;&#x6291;&#x5236; VEGF &#x8BF1;&#x5BFC;&#x4E3B;&#x52A8;&#x8109;&#x73AF;&#x53D1;&#x82BD;&#xFF0C;&#x8868;&#x660E;&#x6297; RPL29 &#x6218;&#x7565;&#x53EF;&#x80FD;&#x5177;&#x6709;&#x6297;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x7684;&#x6F5C;&#x529B;&#x3002;&#x603B;&#x4F53;&#x800C;&#x8A00;&#xFF0C;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x786E;&#x5B9A;&#x8FD9;&#x4E00;&#x635F;&#x5931;&#x6216;&#x635F;&#x8017;&#x7684; RPL29 &#x53EF;&#x4EE5;&#x51CF;&#x5C11;&#x4F53;&#x5185;&#x548C;&#x4F53;&#x5916;&#x7684;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x3002;

&#x5185;&#x6E90;&#x6027;&#x6838;&#x7CD6;&#x4F53;&#x86CB;&#x767D; L29 (RPL29)&#xFF1A; &#x5728;&#x5C0F;&#x9F20;&#x4E2D;&#x7684;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x65B0;&#x53D1;&#x73B0;&#x7684;&#x8C03;&#x8282;&#x5668;&#x3002;

Cellular ribosomal protein L29 (RPL29) is known to be important in protein synthesis, but its function during angiogenesis has never been described before. We have shown previously that mice lacking Î²3-integrins support enhanced tumour angiogenesis and, therefore, deletion of endothelial Î±vÎ²3 can provide a method for discovery of novel regulators of tumour angiogenesis. Here, we describe significant upregulation of RPL29 in Î²3-null endothelial cells at both the mRNA and protein level. Ex vivo, we show that VEGF-stimulated microvessel sprouting was reduced significantly in Rpl29-heterozygous and Rpl29-null aortic ring assays compared with wild-type controls. Moreover, we provide in vivo evidence that RPL29 can regulate tumour angiogenesis. Tumour blood vessel density in subcutaneously grown Lewis lung carcinomas was reduced significantly in Rpl29-mutant mice. Additionally, depletion of Rpl29 using RNA interference inhibited VEGF-induced aortic ring sprouting, suggesting that anti-RPL29 strategies might have anti-angiogenic potential. Overall, our results identify that loss or depletion of RPL29 can reduce angiogenesis in vivo and ex vivo.

Endogenous ribosomal protein L29 (RPL29): a newly identified regulator of angiogenesis in mice.

Prot&eacute;ines ribosomiques cellulaires L29 (RPL29) est connue pour &ecirc;tre importante dans la synth&egrave;se des prot&eacute;ines, mais sa fonction au cours de l&#39;angiogen&egrave;se n&#39;a jamais &eacute;t&eacute; d&eacute;crite avant. Nous avons d&eacute;j&agrave; montr&eacute; que les souris d&eacute;pourvues de &beta;3-int&eacute;grines soutiennent l&#39;angiogen&egrave;se tumorale am&eacute;lior&eacute;e et, donc, la suppression de &alpha;v&beta;3 endoth&eacute;liale peut fournir une m&eacute;thode pour la d&eacute;couverte de nouveaux r&eacute;gulateurs de l&#39;angiogen&egrave;se tumorale. Nous d&eacute;crivons ici upregulation significative de RPL29 dans les cellules endoth&eacute;liales &beta;3 nulle au niveau de l&#39;ARNm et de prot&eacute;ines. Ex vivo, nous montrons que stimul&eacute;e par le VEGF microvaisseaux de germination a &eacute;t&eacute; significativement r&eacute;duite dans les essais d&#39;anneau aortique h&eacute;t&eacute;rozygote-Rpl29 et Rpl29-null par rapport aux t&eacute;moins de type sauvage. En outre, nous fournissons des preuves in vivo que RPL29 peut r&eacute;glementer l&#39;angiogen&egrave;se tumorale. Densit&eacute; des vaisseaux sanguins tumoraux dans cultiv&eacute; par voie sous-cutan&eacute;e Lewis carcinomes du poumon a &eacute;t&eacute; significativement r&eacute;duite chez les souris mutantes Rpl29. En outre, appauvrissement de la couche de Rpl29 &agrave; l&#39;aide de l&#39;interf&eacute;rence ARN inhibe anneau aortique induite par le VEGF la germination, ce qui sugg&egrave;re que les strat&eacute;gies anti-RPL29 pourraient avoir potentiel anti-angiog&eacute;nique. Dans l&#39;ensemble, nos r&eacute;sultats indiquent que la perte ou &eacute;puisement des RPL29 peut r&eacute;duire l&#39;angiogen&egrave;se in vivo et ex vivo.

Prot&eacute;ine ribosomique endog&egrave;ne L29 (RPL29): un r&eacute;gulateur nouvellement identifi&eacute; de l&#39;angiogen&egrave;se chez les souris.

Zellul&auml;re ribosomal Protein L29 (RPL29) ist bekannt, dass die Proteinsynthese wichtig sein, aber seine Funktion w&auml;hrend der Angiogenese ist nie vor beschrieben worden. Wir haben bereits gezeigt, dass mit M&auml;usen fehlt &beta;3-Integrine erweiterte Tumor-Angiogenese unterst&uuml;tzen und daher L&ouml;schung der endothelial &alpha;v&beta;3 eine Methode zur Entdeckung neuartiger Regler der Tumor-Angiogenese bieten. Hier beschreiben wir erhebliche Hochregulation von RPL29 in Endothelzellen &beta;3-Null auf der mRNA und Protein-Ebene. Ex Vivo zeigen wir, dass Keime von VEGF-stimuliert Microvessel in Rpl29-heterozygote und Rpl29-Null Aorten-Ring-Assays, die im Vergleich zu Wildtyp Steuerelemente deutlich reduziert wurde. Dar&uuml;ber hinaus bieten wir in-vivo Beweis, dass RPL29 Tumor-Angiogenese regulieren k&ouml;nnen. Tumor-Blutgef&auml;&szlig;-Dichte in subkutan gewachsen Lewis Lunge Karzinome in Rpl29-Mutanten M&auml;use deutlich reduziert. Zus&auml;tzlich hemmte Ersch&ouml;pfung der Rpl29 mithilfe der RNA-Interferenz, VEGF-induzierte Aorten Ring sprie&szlig;en, darauf hindeutet, dass Anti-RPL29 Strategien Anti-angiogenen Potential haben k&ouml;nnte. Insgesamt unsere Ergebnisse identifizieren dieser Verlust oder Ersch&ouml;pfung der RPL29 verringern Angiogenesis in vivo und ex Vivo.

Endogene ribosomal Protein L29 (RPL29): eine neu identifizierte Regulator der Angiogenese bei M&auml;usen.

Proteina ribosomiale cellulare L29 (RPL29) &egrave; conosciuto per essere importante nella sintesi proteica, ma la sua funzione durante l&#39;angiogenesi non &egrave; mai stata descritta prima. Abbiamo dimostrato in precedenza che topi privi di &beta;3 integrine supportano l&#39;angiogenesi tumorale avanzata e, quindi, eliminazione di &alpha;v&beta;3 endoteliale pu&ograve; fornire un metodo per la scoperta di nuovi regolatori dell&#39;angiogenesi tumorale. Qui, descriviamo significativo sovraregolazione di RPL29 in cellule endoteliali &beta;3-null a livello del mRNA e la proteina. Ex vivo, mostriamo che VEGF-stimolato microvessel spuntano &egrave; stato ridotto significativamente eterozigoti-Rpl29 e Rpl29-null dosaggi anello aortico rispetto ai controlli wild-type. Inoltre, forniamo prove in vivo che RPL29 pu&ograve; regolare l&#39;angiogenesi tumorale. Densit&agrave; dei vasi sanguigni tumorali in coltivato per via sottocutanea Lewis carcinomi del polmone &egrave; stato ridotto significativamente nel topo mutante Rpl29. Inoltre, deplezione di Rpl29 mediante RNA interference inibito indotta da VEGF aortica anello che spuntano, suggerendo strategie anti-RPL29 che potrebbero avere una potenziale anti-angiogenico. Nel complesso, i nostri risultati identificano tale perdita o esaurimento delle RPL29 in grado di ridurre l&#39;angiogenesi in vivo ed ex vivo.

Endogena proteina ribosomiale L29 (RPL29): un regolatore recentemente identificato dell&#39;angiogenesi nei topi.

&#x30BB;&#x30EB;&#x30E9;&#x30FC; &#x30EA;&#x30DC;&#x30BD;&#x30FC;&#x30E0;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA; L29 (RPL29) &#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306E;&#x5408;&#x6210;&#x306B;&#x91CD;&#x8981;&#x3067;&#x3042;&#x308B;&#x3068;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x305D;&#x306E;&#x95A2;&#x6570;&#x306E;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x306E;&#x4E2D;&#x306B;&#x306F;&#x3001;&#x6C7A;&#x3057;&#x3066;&#x524D;&#x306B;&#x8A18;&#x8F09;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306F;&#x4EE5;&#x524D; &beta;3 &#x30A4;&#x30F3;&#x30C6;&#x30B0;&#x30EA;&#x30F3;&#x306B;&#x6B20;&#x3051;&#x3066;&#x3044;&#x308B;&#x30DE;&#x30A6;&#x30B9; &#x30B5;&#x30DD;&#x30FC;&#x30C8;&#x5F37;&#x5316;&#x3055;&#x308C;&#x305F;&#x816B;&#x760D;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3068;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;&#x5185;&#x76AE; &alpha;v&beta;3 &#x306E;&#x524A;&#x9664;&#x816B;&#x760D;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x306E;&#x65B0;&#x3057;&#x3044;&#x8ABF;&#x7BC0;&#x306E;&#x767A;&#x898B;&#x306E;&#x305F;&#x3081;&#x306E;&#x30E1;&#x30BD;&#x30C3;&#x30C9;&#x3092;&#x63D0;&#x4F9B;&#x3067;&#x304D;&#x307E;&#x3059;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;RPL29 &beta;3 null &#x5185;&#x76AE;&#x7D30;&#x80DE; mRNA &#x304A;&#x3088;&#x3073;&#x86CB;&#x767D;&#x8CEA;&#x30EC;&#x30D9;&#x30EB;&#x3067;&#x306E;&#x91CD;&#x5927;&#x306A;&#x30A2;&#x30C3;&#x30D7;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306B;&#x3064;&#x3044;&#x3066;&#x8AAC;&#x660E;&#x3057;&#x307E;&#x3059;&#x3002;&#x524D;&#x306E;&#x30F4;&#x30A3;&#x30F4;&#x30A9;&#x91CE;&#x751F;&#x578B;&#x30B3;&#x30F3;&#x30C8;&#x30ED;&#x30FC;&#x30EB;&#x306B;&#x6BD4;&#x3079;&#x3066; Rpl29 &#x30D8;&#x30C6;&#x30ED;&#x63A5;&#x5408;&#x4F53;&#x3068; Rpl29 null &#x306E;&#x5927;&#x52D5;&#x8108;&#x30EA;&#x30F3;&#x30B0; &#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x5185;&#x306B;&#x5FAE;&#x5C0F;&#x8840;&#x7BA1;&#x306E; VEGF &#x523A;&#x6FC0;&#x767A;&#x82BD;&#x6709;&#x610F;&#x306B;&#x6E1B;&#x5C11;&#x3057;&#x305F;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x307E;&#x305F;&#x3001;RPL29 &#x816B;&#x760D;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3092;&#x8ABF;&#x7BC0;&#x3067;&#x304D;&#x308B; in vivo &#x3067;&#x306E;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;&#x816B;&#x760D;&#x8840;&#x7BA1;&#x5BC6;&#x5EA6;&#x3067;&#x306F;&#x76AE;&#x4E0B;&#x30EB;&#x30A4;&#x30B9;&#x80BA;&#x764C; Rpl29 &#x7A81;&#x7136;&#x5909;&#x7570;&#x4F53;&#x30DE;&#x30A6;&#x30B9;&#x3067;&#x5927;&#x5E45;&#x306B;&#x6E1B;&#x5C11;&#x3057;&#x305F;&#x6210;&#x9577;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;Rpl29 &#x306E; RNA &#x5E72;&#x6E09;&#x3092;&#x7528;&#x3044;&#x305F;&#x67AF;&#x6E07; VEGF &#x8A98;&#x767A;&#x5927;&#x52D5;&#x8108;&#x30EA;&#x30F3;&#x30B0;&#x767A;&#x82BD;&#x3001;&#x30A2;&#x30F3;&#x30C1; RPL29 &#x6226;&#x7565;&#x6297;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x306E;&#x6F5C;&#x5728;&#x6027;&#x3092;&#x6709;&#x3059;&#x308B;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x793A;&#x5506;&#x3092;&#x963B;&#x5BB3;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x5168;&#x4F53;&#x7684;&#x306B;&#x3001;&#x6211;&#x3005; &#x306E;&#x7D50;&#x679C;&#x306F;&#x305D;&#x306E;&#x640D;&#x5931;&#x3092;&#x8B58;&#x5225;&#x307E;&#x305F;&#x306F; RPL29 &#x306E;&#x67AF;&#x6E07;&#x751F;&#x4F53;&#x5185;&#x304A;&#x3088;&#x3073;&#x524D;&#x306E;&#x30F4;&#x30A3;&#x30F4;&#x30A9;&#x306E;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3092;&#x6E1B;&#x3089;&#x3059;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;

&#x5185;&#x56E0;&#x6027;&#x30EA;&#x30DC;&#x30BD;&#x30FC;&#x30E0;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA; L29 (RPL29): &#x30DE;&#x30A6;&#x30B9;&#x306B;&#x304A;&#x3051;&#x308B;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x306E;&#x65B0;&#x305F;&#x306B;&#x540C;&#x5B9A;&#x3055;&#x308C;&#x305F;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30BF;&#x3002;

&#xC140;&#xB8F0;&#xB7EC; ribosomal &#xB2E8;&#xBC31;&#xC9C8; L29 (RPL29) &#xB2E8;&#xBC31;&#xC9C8; &#xD569;&#xC131;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xAC83;&#xC73C;&#xB85C; &#xC54C;&#xB824;&#xC838; &#xC788;&#xC9C0;&#xB9CC; &#xC2E0;&#xC0DD; &#xB3D9;&#xC548; &#xAE30;&#xB2A5; &#xC804;&#xC5D0; &#xACB0;&#xCF54; &#xC124;&#xBA85; &#xB418;&#xC5B4; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &Beta;3 integrins &#xBD80;&#xC871; &#xD55C; &#xB9C8;&#xC6B0;&#xC2A4; &#xC9C0;&#xC6D0; &#xD5A5;&#xC0C1; &#xB41C; &#xC885;&#xC591; &#xD608;&#xAD00; &#xC2E0;&#xC0DD; &#xBC0F; &#xB0B4; &#xD53C; &alpha;v&beta;3 &#xC0AD;&#xC81C; &#xC885;&#xC591; &#xD608;&#xAD00; &#xC2E0;&#xC0DD;&#xC758; &#xC18C;&#xC124; &#xB808; &#xADE4; &#xB808;&#xC774; &#xD130;&#xC758; &#xBC1C;&#xACAC;&#xC5D0; &#xB300; &#xD55C; &#xBA54;&#xC11C;&#xB4DC;&#xB97C; &#xC81C;&#xACF5;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xB530;&#xB77C;&#xC11C;, &#xC6B0;&#xB9AC;&#xB294; &#xC774;&#xC804;&#xC5D0; &#xD45C;&#xC2DC; &#xB429;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xB294; &#xC0C1;&#xB2F9;&#xD55C; upregulation Mrna&#xC640; &#xB2E8;&#xBC31;&#xC9C8; &#xC218;&#xC900;&#xC5D0;&#xC11C; null&#xC774; &beta;3 &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; rpl29&#xC758; &#xC124;&#xBA85;. Ex vivo, &#xC6B0;&#xB9AC; microvessel VEGF &#xC790;&#xADF9; &#xB3CB; &#xC544; &#xC57C;&#xC0DD;-&#xD0C0;&#xC785; &#xCEE8;&#xD2B8;&#xB864;&#xC5D0; &#xBE44;&#xD574; Rpl29 heterozygous &#xBC0F; Rpl29 null &#xB300;&#xB3D9;&#xB9E5; &#xBC18;&#xC9C0; &#xBD84;&#xC11D; &#xC2E4;&#xD5D8;&#xC5D0;&#xC11C; &#xD06C;&#xAC8C; &#xAC10;&#xC18C; &#xD588;&#xB2E4; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xC6B0;&#xB9AC; RPL29 &#xC885;&#xC591; &#xD608;&#xAD00; &#xC2E0;&#xC0DD;&#xC744; &#xC870;&#xC808; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; vivo&#xC5D0;&#xC11C; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xC885;&#xC591; &#xD608;&#xAD00; &#xBC00;&#xB3C4; subcutaneously &#xD3D0; carcinomas Rpl29 &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC0DD;&#xC950;&#xC5D0;&#xC11C; &#xD06C;&#xAC8C; &#xAC10;&#xC18C; &#xD588;&#xB2E4; &#xB8E8;&#xC774;&#xC2A4; &#xC131;&#xC7A5;. &#xB610;&#xD55C;, &#xAC04;&#xC12D; RNA&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; rpl29&#xC758; &#xACE0;&#xAC08; &#xC800;&#xD574; &#xB300;&#xB3D9;&#xB9E5; &#xBC18;&#xC9C0; VEGF &#xC720;&#xB3C4; &#xB3CB; &#xC544;, &#xC548;&#xD2F0; RPL29 &#xC804;&#xB7B5; &#xC548;&#xD2F0; angiogenic &#xC7A0;&#xC7AC;&#xB825; &#xC788;&#xB294; &#xC81C;&#xC548;. &#xC804;&#xBC18;&#xC801;&#xC73C;&#xB85C;, &#xC6B0;&#xB9AC;&#xC758; &#xACB0;&#xACFC; &#xC2DD;&#xBCC4; &#xC190;&#xC2E4; &#xB610;&#xB294; rpl29&#xC758; &#xACE0;&#xAC08; &#xC2E0;&#xC0DD; &#xC0DD;&#xCCB4; &#xC870;&#xAC74; &#xBC0F; &#xBE44;&#xBCF4; &#xC804;&#xC744; &#xC904;&#xC77C; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xC0DD; ribosomal &#xB2E8;&#xBC31;&#xC9C8; L29 (RPL29): &#xC950;&#xC5D0; &#xC2E0;&#xC0DD;&#xC758; &#xC0C8;&#xB85C; &#xD655;&#xC778;&#xB41C; &#xB41C; &#xB808; &#xADE4; &#xB808;&#xC774; &#xD130;.

Prote&iacute;nas celulares L29 (RPL29) es conocido por ser importante en la s&iacute;ntesis de prote&iacute;nas, pero su funci&oacute;n durante la angiog&eacute;nesis nunca se ha descrito antes. Hemos demostrado previamente que ratones que carecen de integrinas &beta;3 angiog&eacute;nesis tumoral mayor y, por lo tanto, la eliminaci&oacute;n de &alpha;v&beta;3 endotelial puede proporcionar un m&eacute;todo para el descubrimiento de nuevos reguladores de la angiog&eacute;nesis tumoral. Aqu&iacute;, describimos upregulation significativo de RPL29 en c&eacute;lulas endoteliales &beta;3 nulo nivel tanto el mRNA y la prote&iacute;na. Ex vivo, mostramos que VEGF-estimulada microvasos brotaci&oacute;n se redujo significativamente en ensayos de anillo a&oacute;rtico Rpl29-heterozig&oacute;tica y Rpl29 nulo en comparaci&oacute;n con controles de tipo salvaje. Adem&aacute;s, Proporcionamos evidencia in vivo que RPL29 puede regular la angiog&eacute;nesis tumoral. Densidad de los vasos sangu&iacute;neos del tumor en forma subcut&aacute;nea crecido Lewis carcinomas pulmonares se redujo significativamente en ratones mutantes Rpl29. Adem&aacute;s, agotamiento de Rpl29 utilizando ARN de interferencia inhibe anillo a&oacute;rtico VEGF inducida por brotaci&oacute;n, sugiriendo que anti-RPL29 estrategias podr&iacute;an tener potencial anti-angiog&eacute;nica. En general, nuestros resultados identifican esa p&eacute;rdida o agotamiento de RPL29 puede reducir la angiog&eacute;nesis in vivo como ex vivo.

Prote&iacute;nas end&oacute;genas L29 (RPL29): un regulador nuevamente identificado de la angiog&eacute;nesis en ratones.

Although microRNAs (miRNA) have been broadly studied in cancer, comparatively less is understood about their role in progression. Here we report that miR-153 has a dual role during progression of colorectal cancer by enhancing cellular invasiveness and platinum-based chemotherapy resistance. miRNA profiling revealed that miR-153 was highly expressed in a cellular model of advanced stage colorectal cancer. Its upregulation was also noted in primary human colorectal cancer compared with normal colonic epithelium and in more advanced colorectal cancer stages compared with early stage disease. In colorectal cancer patients followed for 50 months, 21 of 30 patients with high levels of miR-153 had disease progression compared with others in this group with low levels of miR-153. Functional studies revealed that miR-153 upregulation increased colorectal cancer invasiveness and resistance to oxaliplatin and cisplatin both in vitro and in vivo. Mechanistic investigations indicated that miR-153 promoted invasiveness indirectly by inducing matrix metalloprotease enzyme 9 production, whereas drug resistance was mediated directly by inhibiting the Forkhead transcription factor Forkhead box O3a (FOXO3a). In support of the latter finding, we found that levels of miR-153 and FOXO3a were inversely correlated in matched human colorectal cancer specimens. Our findings establish key roles for miR-153 overexpression in colorectal cancer progression, rationalizing therapeutic strategies to target expression of this miRNA for colorectal cancer treatment.

miR-153 supports colorectal cancer progression via pleiotropic effects that enhance invasion and chemotherapeutic resistance.

Hematopoietic stem and progenitor cells (HSPCs) are exposed to low levels of oxygen in the bone marrow niche, and hypoxia-inducible factors (HIFs) are the main regulators of cellular responses to oxygen variation. Recent studies using conditional knockout mouse models have unveiled a major role for HIF-1α in the maintenance of murine HSCs; however, the role of HIF-2α is still unclear. Here, we show that knockdown of HIF-2α, and to a much lesser extent HIF-1α, impedes the long-term repopulating ability of human CD34(+) umbilical cord blood cells. HIF-2α-deficient HSPCs display increased production of reactive oxygen species (ROS), which subsequently stimulates endoplasmic reticulum (ER) stress and triggers apoptosis by activation of the unfolded-protein-response (UPR) pathway. HIF-2α deregulation also significantly decreased engraftment ability of human acute myeloid leukemia (AML) cells. Overall, our data demonstrate a key role for HIF-2α in the maintenance of human HSPCs and in the survival of primary AML cells.

Richard Mitter

Bioinformatics Unit

London Research Institute, Cancer Research UK

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

Richard Mitter

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

London Research Institute, Cancer Research UK

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts

Bioinformatics Unit