Reusable Single Cell for Iterative Epigenomic Analyses

Current single-cell epigenome analyses are designed for single use. The cell is discarded after a single use, preventing analysis of multiple epigenetic ...

National Institutes of Health

&#x5728;&#x5F00;&#x53D1;&#x671F;&#x95F4;&#xFF0C;&#x4EE5;&#x53CA;&#x5728;&#x6210;&#x719F;&#x7684;&#x54FA;&#x4E73;&#x52A8;&#x7269;&#x4E2D;&#x7684;&#x65B0;&#x8239;&#x5F62;&#x6210;&#x4E00;&#x822C;&#x6536;&#x76CA;&#x901A;&#x8FC7;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x3002;&#x867D;&#x7136;&#x5404;&#x79CD;&#x5206;&#x5B50;&#x548C;&#x4FE1;&#x53F7;&#x4F20;&#x5BFC;&#x901A;&#x8DEF;&#x5DF2;&#x77E5;&#x9020;&#x6210;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x53D1;&#x82BD;&#x548C;&#x91CD;&#x5851;&#x5728;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x8FC7;&#x7A0B;&#xFF0C;&#x8FD9;&#x4E00;&#x590D;&#x6742;&#x8FC7;&#x7A0B;&#x7684;&#x8BB8;&#x591A;&#x65B9;&#x9762;&#x4ECD;&#x7136;&#x4E0D;&#x660E;&#x539F;&#x56E0;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x663E;&#x793A;&#xFF0C;&#x8DE8;&#x819C;&#x7684; semaphorin6A (Sema6A) &#x8868;&#x793A;&#x5185;, &#x76AE;&#x7EC6;&#x80DE;&#xFF0C;&#x5E76;&#x4E14;&#x901A;&#x8FC7;&#x8C03;&#x8282;&#x8868;&#x8FBE;&#x53CA; VEGFR2&#xFF0C;&#x5373;&#x7EF4;&#x6301;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x6D3B;&#x6027;&#x7684;&#x81EA;&#x5206;&#x6CCC;&#x751F;&#x957F;&#x56E0;&#x5B50;&#x53D7;&#x4F53;&#x4FE1;&#x53F7;&#x7684;&#x4FE1;&#x53F7;&#x8C03;&#x8282;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x5B58;&#x6D3B;&#x548C;&#x751F;&#x957F;&#x3002;&#x6C89;&#x9ED8; Sema6A &#x4E3B;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x4FC3;&#x8FDB;&#x7EC6;&#x80DE;&#x6B7B;&#x4EA1;&#xFF0C;&#x4E0D;&#x7531;&#x5916;&#x6E90;&#x6551;&#x51FA; VEGF A &#x6216; FGF2&#xFF0C;&#x5F52;&#x56E0;&#x4E8E;&#x6765;&#x81EA;&#x5185;&#x6E90;&#x6027; VEGF &#x7684; prosurvival &#x4FE1;&#x53F7;&#x635F;&#x5931;&#x3002;&#x5C0F;&#x9F20;&#x7EC4;&#x7EC7;&#x7684;&#x5206;&#x6790;&#x8868;&#x660E; Sema6A &#x8868;&#x793A;&#x5728;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x548C;&#x91CD;&#x5851;&#x8239;&#x53EA;&#x3002;&#x5728;&#x906E;&#x65AD;&#x8239;&#x53EA;&#x589E;&#x52A0;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x6B7B;&#x4EA1;&#xFF0C;&#x4E0E;&#x76F8;&#x5173;&#x7684;&#x590D;&#x6742;&#x6027;&#x548C;&#x5F02;&#x5E38;&#x964D;&#x4F4E;&#x7684;&#x89C6;&#x7F51;&#x819C;&#x8840;&#x7BA1;&#x53D1;&#x5C55; Sema6A &#x7A7A;&#x7A81;&#x53D8;&#x5C0F;&#x9F20;&#x8868;&#x73B0;&#x51FA;&#x91CD;&#x5927;&#x7F3A;&#x9677;&#x3002;&#x6210;&#x4EBA; Sema6A null &#x5C0F;&#x9F20;&#x8868;&#x73B0;&#x51FA;&#x51CF;&#x5C11;&#x7684;&#x80BF;&#x7624;&#x3001; matrigel &#x548C;&#x8109;&#x7EDC;&#x819C;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x4E0E;&#x63A7;&#x4EF6;&#x76F8;&#x6BD4;&#x3002;Sema6A &#x5728;&#x4E2D;&#x67A2;&#x795E;&#x7ECF;&#x7CFB;&#x7EDF;&#x7684;&#x53D1;&#x5C55;&#x4E2D;&#x626E;&#x6F14;&#x91CD;&#x8981;&#x89D2;&#x8272;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x5C55;&#x793A;&#x5B83;&#x8FD8;&#x89C4;&#x5B9A;&#x4E86;&#x8840;&#x7BA1;&#x53D1;&#x80B2;&#x4E0E;&#x6210;&#x4EBA;&#x7684;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x3002;

Semaphorin 6A &#x901A;&#x8FC7;&#x8C03;&#x8282; VEGF &#x4FE1;&#x53F7;&#x8C03;&#x8282;&#x8840;&#x7BA1;&#x751F;&#x6210;&#x3002;

Formation of new vessels during development and in the mature mammal generally proceeds through angiogenesis. Although a variety of molecules and signaling pathways are known to underlie endothelial cell sprouting and remodeling during angiogenesis, many aspects of this complex process remain unexplained. Here we show that the transmembrane semaphorin6A (Sema6A) is expressed in endothelial cells, and regulates endothelial cell survival and growth by modulating the expression and signaling of VEGFR2, which is known to maintain endothelial cell viability by autocrine VEGFR signaling. The silencing of Sema6A in primary endothelial cells promotes cell death that is not rescued by exogenous VEGF-A or FGF2, attributable to the loss of prosurvival signaling from endogenous VEGF. Analyses of mouse tissues demonstrate that Sema6A is expressed in angiogenic and remodeling vessels. Mice with null mutations of Sema6A exhibit significant defects in hyaloid vessels complexity associated with increased endothelial cell death, and in retinal vessels development that is abnormally reduced. Adult Sema6A-null mice exhibit reduced tumor, matrigel, and choroidal angiogenesis compared with controls. Sema6A plays important roles in development of the nervous system. Here we show that it also regulates vascular development and adult angiogenesis.

Semaphorin 6A regulates angiogenesis by modulating VEGF signaling.

Formation de nouveaux vaisseaux au cours du d&eacute;veloppement et chez le mammif&egrave;re mature produit g&eacute;n&eacute;ralement par le biais de l&#39;angiogen&egrave;se. Bien qu&#39;une vari&eacute;t&eacute; de mol&eacute;cules et les voies de signalisation sont connus pour sous-tendre le bourgeonnement des cellules endoth&eacute;liales et remodelage au cours de l&#39;angiogen&egrave;se, les nombreux aspects de ce processus complexe demeurent inexpliqu&eacute;s. Ici, nous montrons que la semaphorin6A transmembranaires (Sema6A) est exprim&eacute; dans les cellules endoth&eacute;liales et r&eacute;gule la croissance et la survie des cellules endoth&eacute;liales par modulation de l&#39;expression et la signalisation de VEGFR2, qui est connu pour maintenir la viabilit&eacute; des cellules endoth&eacute;liales par autocrine VEGFR signalisation. Le silence des Sema6A dans les cellules endoth&eacute;liales primaires favorise la mort cellulaire qui n&#39;est pas sauv&eacute;e par exog&egrave;ne VEGF-A ou FGF2, attribuable &agrave; la perte de signalisation apoptotique de VEGF endog&egrave;ne. Analyses des tissus de souris d&eacute;montrent que Sema6A est exprim&eacute;e dans les vaisseaux angiog&eacute;niques et le remodelage. Souris avec des mutations nulles de Sema6A pr&eacute;sentent des d&eacute;fauts importants par navires hyalo&iuml;de complexit&eacute; associ&eacute;e &agrave; la mort de cellules endoth&eacute;liales et dans le d&eacute;veloppement de vaisseaux r&eacute;tiniens qui est anormalement r&eacute;duit. Chez la souris adulte Sema6A null pi&egrave;ce r&eacute;duction tumorale, matrigel et angiogen&egrave;se choro&iuml;dienne compar&eacute;e aux t&eacute;moins. Sema6A joue un r&ocirc;le important dans le d&eacute;veloppement du syst&egrave;me nerveux. Nous d&eacute;montrons qu&#39;il r&eacute;glemente &eacute;galement d&eacute;veloppement vasculaire et angiogen&egrave;se adulte.

S&eacute;maphorine 6 a r&eacute;glemente l&#39;angiogen&egrave;se en modulant la signalisation du VEGF.

Bildung neuer Schiffe w&auml;hrend der Entwicklung und in der Reife S&auml;ugetier durchl&auml;uft in der Regel Angiogenesis. Obwohl eine Vielzahl von Molek&uuml;len und Signalwege sind bekannterma&szlig;en Endothelzellen sprie&szlig;en und Umbau w&auml;hrend Angiogenesis zugrunde liegen, sind viele Aspekte dieses komplexen Prozesses ungekl&auml;rt. Hier zeigen wir, dass die Transmembran-semaphorin6A (Sema6A) wird ausgedr&uuml;ckt in Endothelzellen und regelt endothelial Zellen &uuml;berleben und Wachstum durch Modulierung des Ausdrucks und Signalisierung der VEGFR2, die durch das Autocrine signalisieren VEGFR, endothelial Zelle Entwicklungsf&auml;higkeit beizubehalten bekannt ist. Der Unterdr&uuml;ckung der Sema6A im prim&auml;ren Endothelzellen f&ouml;rdert Zelltod, die nicht durch exogene gerettet wird VEGF-A oder FGF2, auf den Verlust der prosurvival Signalisierung von endogenen VEGF zur&uuml;ckzuf&uuml;hren. Analysen der Maus Gewebe zeigen, dass Sema6A in der Angiogenese und Umbau von Schiffen ausgedr&uuml;ckt wird. M&auml;use mit null Mutationen des Sema6A weisen erhebliche M&auml;ngel im hyaloid Schiffe Komplexit&auml;t im Zusammenhang mit erh&ouml;hten endothelial Zellentod und retinale Gef&auml;&szlig;e-Entwicklung, die ungew&ouml;hnlich reduziert wird. Erwachsene Sema6A-Null-M&auml;use zeigen reduzierten Tumor, Matrigel und Choroidale Angiogenese im Vergleich zu Kontrollen. Sema6A spielt eine wichtige Rolle in der Entwicklung des Nervensystems. Hier zeigen wir, dass es auch vaskul&auml;re Entwicklung und Erwachsenen Angiogenese reguliert.

Semaphorine 6A regelt die Angiogenese durch Modulation VEGF signalisieren.

Formazione di nuovi vasi durante lo sviluppo e il mammifero maturo generalmente procede attraverso l&#39;angiogenesi. Anche se una variet&agrave; di molecole e vie di segnalazione sono alla base delle cellule endoteliali che spuntano e rimodellamento durante l&#39;angiogenesi, molti aspetti di questo complesso processo restano inspiegabili. Qui vi mostriamo che la semaphorin6A del transmembrane (Sema6A) &egrave; espressa nelle cellule endoteliali e regola la crescita e la sopravvivenza delle cellule endoteliali modulando l&#39;espressione e la segnalazione di VEGFR2, che &egrave; noto per mantenere la vitalit&agrave; delle cellule endoteliali con segnalazione di autocrine VEGFR. Il silenziamento di Sema6A in cellule endoteliali primarie promuove la morte delle cellule che non viene salvato da esogeno VEGF-A o FGF2, attribuibile alla perdita di prosurvival segnalazione dal VEGF endogeno. Analisi dei tessuti del mouse dimostrano che Sema6A &egrave; espresso nei vasi angiogenica e rimodellamento. Topi con mutazioni null di Sema6A presentano difetti significativi nella complessit&agrave; hyaloid vasi associati a morte aumentato delle cellule endoteliali e nello sviluppo di vasi della retina che &egrave; anormalmente ridotta. Topi adulti Sema6A-null esibiscono tumore ridotto e matrigel coroidale angiogenesi rispetto ai controlli. Sema6A svolge ruoli importanti nello sviluppo del sistema nervoso. Qui vi mostriamo che regola anche lo sviluppo vascolare e angiogenesi adulto.

Semaphorin 6A regola l&#39;angiogenesi modulando VEGF segnalazione.

&#x6210;&#x719F;&#x54FA;&#x4E73;&#x52D5;&#x7269;&#x3067;&#x3001;&#x958B;&#x767A;&#x6642;&#x306B;&#x65B0;&#x3057;&#x3044;&#x8840;&#x7BA1;&#x306E;&#x5F62;&#x6210;&#x306F;&#x3001;&#x4E00;&#x822C;&#x7684;&#x306B;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3092;&#x901A;&#x3058;&#x3066;&#x9032;&#x884C;&#x3057;&#x307E;&#x3059;&#x3002;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x306E;&#x840C;&#x82BD;&#x3068;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x4E2D;&#x306B;&#x6539;&#x9020;&#x306E;&#x6839;&#x5E95;&#x306B;&#x3042;&#x308B;&#x3001;&#x3055;&#x307E;&#x3056;&#x307E;&#x306A;&#x5206;&#x5B50;&#x3084;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x7D4C;&#x8DEF;&#x3092;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x3053;&#x306E;&#x8907;&#x96D1;&#x306A;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306E;&#x591A;&#x304F;&#x306E;&#x5074;&#x9762;&#x306F;&#x539F;&#x56E0;&#x4E0D;&#x660E;&#x6B8B;&#x308A;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x819C;&#x8CAB;&#x901A; semaphorin6A &#x8868;&#x793A; &#xFF08;Sema6A&#xFF09; &#x5185;&#x76AE;&#x7D30;&#x80DE;&#x3067;&#x767A;&#x73FE;&#x3057;&#x3001;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x306E;&#x751F;&#x5B58;&#x3068;&#x6210;&#x9577;&#x5F0F;&#x3092;&#x5909;&#x8ABF;&#x3057;&#x3066;&#x30AA;&#x30FC;&#x30C8;&#x30AF;&#x30EA;&#x30F3; VEGFR &#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x306B;&#x3088;&#x308B;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x751F;&#x5B58;&#x7387;&#x3092;&#x7DAD;&#x6301;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B; VEGFR2 &#x306E;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;&#x30D7;&#x30E9;&#x30A4;&#x30DE;&#x30EA;&#x306E;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x306B;&#x304A;&#x3051;&#x308B; Sema6A &#x306E;&#x30B5;&#x30A4;&#x30EC;&#x30F3;&#x30B7;&#x30F3;&#x30B0;&#x306B;&#x3088;&#x308B;&#x5916;&#x56E0;&#x6027;&#x306A;&#x3044;&#x304C;&#x6551;&#x51FA;&#x3055;&#x308C;&#x308B;&#x7D30;&#x80DE;&#x6B7B;&#x4FC3;&#x9032; VEGF A &#x307E;&#x305F;&#x306F; FGF2 &#x306E; prosurvival &#x304B;&#x3089;&#x306E;&#x5185;&#x56E0;&#x6027; VEGF &#x30B7;&#x30B0;&#x30CA;&#x30EA;&#x30F3;&#x30B0;&#x306E;&#x640D;&#x5931;&#x306B;&#x8D77;&#x56E0;&#x3057;&#x307E;&#x3059;&#x3002;&#x30DE;&#x30A6;&#x30B9;&#x81D3;&#x5668;&#x306E;&#x5206;&#x6790;&#x306F;&#x3001;Sema6A &#x304C;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3084;&#x6539;&#x9020;&#x306E;&#x8239;&#x8236;&#x3067;&#x8868;&#x3055;&#x308C;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;Sema6A &#x306E; null &#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x3092;&#x6301;&#x3064;&#x30DE;&#x30A6;&#x30B9;&#x5897;&#x52A0;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x6B7B;&#x3068;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x785D;&#x5B50;&#x4F53;&#x8840;&#x7BA1;&#x306E;&#x8907;&#x96D1;&#x3055;&#x3068;&#x7570;&#x5E38;&#x6E1B;&#x5C11;&#x7DB2;&#x819C;&#x8840;&#x7BA1;&#x306E;&#x958B;&#x767A;&#x306E;&#x91CD;&#x5927;&#x306A;&#x6B20;&#x9665;&#x3092;&#x5C55;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x5927;&#x4EBA;&#x306E; Sema6A null &#x30DE;&#x30A6;&#x30B9;&#x524A;&#x6E1B;&#x816B;&#x760D;&#x3001;&#x30DE;&#x30C8;&#x30EA;&#x30B2;&#x30EB;&#x3001;&#x304A;&#x3088;&#x3073;&#x8108;&#x7D61;&#x819C;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x5236;&#x5FA1;&#x3068;&#x6BD4;&#x8F03;&#x5C55;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;Sema6A &#x306F;&#x795E;&#x7D4C;&#x7CFB;&#x306E;&#x767A;&#x9054;&#x306B;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x3092;&#x679C;&#x305F;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x305D;&#x308C;&#x306F;&#x307E;&#x305F;&#x8840;&#x7BA1;&#x306E;&#x958B;&#x767A;&#x3068;&#x6210;&#x4EBA;&#x306E;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3092;&#x8ABF;&#x6574;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

&#x30BB;&#x30DE;&#x30D5;&#x30A9;&#x30EA;&#x30F3; 6 a VEGF &#x3092;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x3092;&#x5909;&#x8ABF;&#x306B;&#x8840;&#x7BA1;&#x65B0;&#x751F;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;

&#xAC1C;&#xBC1C; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xBC0F; &#xC131;&#xC219;&#xD55C; &#xD3EC;&#xC720;&#xB958;&#xC5D0; &#xC0C8;&#xB85C;&#xC6B4; &#xD608;&#xAD00;&#xC758; &#xD615;&#xC131;&#xC740; &#xC77C;&#xBC18;&#xC801;&#xC73C;&#xB85C; &#xC2E0;&#xC0DD; &#xD608;&#xAD00;&#xC744; &#xD1B5;&#xD574; &#xC9C4;&#xD589; &#xB429;&#xB2C8;&#xB2E4;. &#xB2E4;&#xC591; &#xD55C; &#xBD84;&#xC790;&#xC640; &#xC2E0;&#xD638; &#xACBD;&#xB85C; &#xAE30;&#xCD08; &#xB0B4; &#xD53C; &#xC138;&#xD3EC; &#xB3CB; &#xC544; &#xBC0F; &#xC2E0;&#xC0DD; &#xC911; &#xB9AC; &#xBAA8;&#xB378;&#xB9C1; &#xC54C;&#xB824;&#xC838; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;, &#xD558;&#xC9C0;&#xB9CC;&#xC774; &#xBCF5;&#xC7A1; &#xD55C; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC758; &#xC5EC;&#xB7EC; &#xCE21;&#xBA74; &#xC124;&#xBA85;&#xD560; &#xC218; &#xC5C6;&#xB294; &#xB0A8;&#xC544; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;&#xB294; &#xB9C9; &#xD6A1;&#xB2E8; semaphorin6A &#xBCF4;&#xC5EC; (Sema6A)&#xC740; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC5D0; &#xD45C;&#xD604; &#xB418; &#xACE0; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC758; &#xC0DD;&#xC874;&#xACFC; &#xC131;&#xC7A5; &#xC2DD; &#xBCC0;&#xC870; &#xBC0F; autocrine VEGFR &#xC2E0;&#xD638;&#xC5D0; &#xC758;&#xD574; &#xB0B4; &#xD53C; &#xC138;&#xD3EC; &#xC0DD;&#xC874; &#xB2A5;&#xB825;&#xC744; &#xC720;&#xC9C0; &#xD558;&#xB294; &#xAC83;&#xC73C;&#xB85C; &#xC54C;&#xB824;&#xC838; &#xC788;&#xB294; vegfr2&#xC758; &#xC2E0;&#xD638;&#xC5D0; &#xC758;&#xD574; &#xC870;&#xC808;. Exogenous &#xC758;&#xD574; &#xAD6C;&#xCD9C; &#xD558;&#xC9C0; &#xC138;&#xD3EC;&#xC758; &#xC8FD;&#xC74C;&#xC744; &#xCD09;&#xC9C4; &#xAE30;&#xBCF8; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; sema6a&#xC758; &#xC785;&#xC744; VEGF &#xB610;&#xB294; FGF2 &#xC0DD; VEGF&#xC5D0;&#xC11C; prosurvival &#xC2E0;&#xD638; &#xC190;&#xC2E4;&#xC5D0; &#xAE30;&#xC778; &#xD569;&#xB2C8;&#xB2E4;. &#xB9C8;&#xC6B0;&#xC2A4; &#xC870;&#xC9C1;&#xC758; &#xBD84;&#xC11D; Sema6A angiogenic &#xBC0F; &#xB9AC; &#xBAA8;&#xB378;&#xB9C1; &#xD608;&#xAD00;&#xC5D0; &#xD45C;&#xD604; &#xD558;&#xB294; &#xBC29;&#xBC95;&#xC744; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. Sema6a&#xC758; null &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;&#xC640; &#xC950; hyaloid &#xC120;&#xBC15; &#xBCF5;&#xC7A1;&#xC131; &#xC99D;&#xAC00; &#xB0B4; &#xD53C; &#xC138;&#xD3EC;&#xC758; &#xC8FD;&#xC74C;&#xC744; &#xC5F0;&#xAD00; &#xBC0F; &#xBE44;&#xC815;&#xC0C1;&#xC801;&#xC73C;&#xB85C; &#xAC10;&#xC18C; &#xB9DD;&#xB9C9; &#xD608;&#xAD00; &#xAC1C;&#xBC1C;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xACB0;&#xD568; &#xC804;&#xC2DC;. &#xC131;&#xC778; Sema6A null &#xC950; &#xC804;&#xC2DC; &#xAC10;&#xC18C; &#xC885;&#xC591;, matrigel, &#xADF8;&#xB9AC;&#xACE0; &#xCEE8;&#xD2B8;&#xB864;&#xC5D0; &#xBE44;&#xD574; &#xC548; &#xC2E0;&#xC0DD;. Sema6A &#xC2E0;&#xACBD; &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xAC1C;&#xBC1C;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xC5ED;&#xD560;&#xC744; &#xD55C;&#xB2E4;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC;&#xB294; &#xADF8;&#xAC83;&#xC740; &#xB610;&#xD55C; &#xD608;&#xAD00; &#xAC1C;&#xBC1C; &#xBC0F; &#xC131;&#xC778; &#xC2E0;&#xC0DD; &#xC870;&#xC808; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;.

Semaphorin 6A VEGF &#xC2E0;&#xD638;&#xB97C; &#xBCC0;&#xC870; &#xD558; &#xC5EC; &#xC2E0;&#xC0DD;&#xC744; &#xC870;&#xC808;.

Formaci&oacute;n de nuevos vasos durante el desarrollo y en los mam&iacute;feros adultos generalmente procede a trav&eacute;s de la angiog&eacute;nesis. Aunque una variedad de mol&eacute;culas y v&iacute;as de se&ntilde;alizaci&oacute;n se sabe que subyacen la brotaci&oacute;n de la c&eacute;lula endotelial y remodelado durante la angiog&eacute;nesis, muchos aspectos de este complejo proceso siguen siendo inexplicables. Aqu&iacute; demostramos que la semaphorin6A transmembrana (Sema6A) se expresa en las c&eacute;lulas endoteliales y regula el crecimiento y la supervivencia de c&eacute;lulas endoteliales por modulaci&oacute;n de la expresi&oacute;n y se&ntilde;alizaci&oacute;n de VEGFR2, que se conoce para mantener la viabilidad de las c&eacute;lulas endoteliales por autocrina VEGFR se&ntilde;alizaci&oacute;n. El silenciamiento de Sema6A en las c&eacute;lulas endoteliales primarias promueve la muerte celular que no es rescatada por ex&oacute;genos VEGF-A o FGF2, atribuibles a la p&eacute;rdida de se&ntilde;alizaci&oacute;n favorecen de VEGF end&oacute;geno. An&aacute;lisis de tejidos de rat&oacute;n demuestran que Sema6A se expresa en los vasos angiog&eacute;nicos y remodelaci&oacute;n. Ratones con mutaciones nulas de Sema6A exhiben defectos importantes en complejidad de vasos hialoide asociadas con la muerte de la c&eacute;lula endotelial creciente y en el desarrollo de vasos retinianos que anormalmente se reduce. Ratones Sema6A nulo adultos exhiben tumor reducido matrigel y angiog&eacute;nesis coroides comparada con controles. Sema6A juega un papel importante en el desarrollo del sistema nervioso. Aqu&iacute; mostramos que tambi&eacute;n regula desarrollo vascular y angiog&eacute;nesis para adultos.

Semaforina 6A regula la angiog&eacute;nesis modulando se&ntilde;alizaci&oacute;n de VEGF.

Myeloid cells that orchestrate malignant progression in the tumor microenvironment offer targets for a generalized strategy to attack solid tumors. Through an analysis of tumor microenvironments, we explored an experimental model of lung cancer that uncovered a network of Dll4/Notch/TGF-β1 signals that links myeloid cells to cancer progression. Myeloid cells attracted to the tumor microenvironment by the tumor-derived cytokines CCL2 and M-CSF expressed increased levels of the Notch ligand Dll4, thereby activating Notch signaling in the tumor cells and amplifying tumor-intrinsic Notch activation. Heightened Dll4/Notch signaling in tumor cells magnified TGF-β-induced pSMAD2/3 signaling and was required to sustain TGF-β-induced tumor cell growth. Conversely, Notch blockade reduced TGF-β signaling and limited lung carcinoma tumor progression. Corroborating these findings, by interrogating RNAseq results from tumor and adjacent normal tissue in clinical specimens of human head and neck squamous carcinoma, we found evidence that TGF-β/Notch crosstalk contributed to progression. In summary, the myeloid cell-carcinoma signaling network we describe uncovers novel mechanistic links between the tumor microenvironment and tumor growth, highlighting new opportunities to target tumors where this network is active.

Tumor-infiltrating myeloid cells activate Dll4/Notch/TGF-β signaling to drive malignant progression.

Cell signals integral to the tumor microenvironment influence cancer progression. Tumor-associated myeloid cells secrete pro-tumorigenic agents including, but not limited to, the potent cytokine transforming growth factor β (TGFβ). We have discovered a network of extrinsic signals including delta-like 4 (Dll4), Notch and TGFβ, linking malignant cells and tumor-infiltrating myeloid cells, a nexus portending a clinically-relevant anticancer treatment.

Notch and TGFβ: Functional partners facilitating tumor progression.

Angiogenesis produces primitive vascular networks that need pruning to yield hierarchically organized and functional vessels. Despite the critical importance of vessel pruning to vessel patterning and function, the mechanisms regulating this process are not clear. Here we show that EphrinB2, a well-known player in angiogenesis, is an essential regulator of endothelial cell death and vessel pruning. This regulation depends upon phosphotyrosine-EphrinB2 signalling repressing c-jun N-terminal kinase 3 activity via STAT1. JNK3 activation causes endothelial cell death. In the absence of JNK3, hyaloid vessel physiological pruning is impaired, associated with abnormal persistence of hyaloid vessels, defective retinal vasculature and microphthalmia. This syndrome closely resembles human persistent hyperplastic primary vitreus (PHPV), attributed to failed involution of hyaloid vessels. Our results provide evidence that EphrinB2/STAT1/JNK3 signalling is essential for vessel pruning, and that defects in this pathway may contribute to PHPV. 

EphrinB2 controls vessel pruning through STAT1-JNK3 signalling.

Interleukin-23 (IL-23), a heterodimeric cytokine composed of the unique p19 peptide (IL-23p19) and a peptide called IL-12p40, which is shared with IL-12, is implicated in Crohn's disease, rheumatoid arthritis, psoriasis, and other immune-mediated inflammatory diseases. Endothelial cells produce the IL-23p19 peptide in the absence of the IL-12p40 chain and thus do not make heterodimeric IL-23. We found that intercellular IL-23p19 increased the cell surface abundances of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) on endothelial cells, which enhanced the attachment of leukocytes and increased their transendothelial migration. Intracellular p19 associated with the cytokine receptor subunit gp130 and stimulated the gp130-dependent activation of signal transducer and activator of transcription 3 (STAT3) signaling. Proinflammatory factors promoted the generation of IL-23p19 in endothelial cells. The adventitial capillaries of inflamed temporal arteries in patients with giant-cell arteritis (GCA) had endothelial p19 protein associated with gp130, but did not contain the IL-12p40 chain. Because adventitial capillaries are essential for the entry of inflammatory cells into arterial walls, these data suggest that p19 may contribute to GCA disease and could represent a therapeutic target. Our results provide evidence that IL-23p19 is a previously unrecognized endothelial proinflammatory peptide that promotes leukocyte transendothelial migration, advancing our current understanding of the complexities of inflammatory responses. 

Identification of IL-23p19 as an endothelial proinflammatory peptide that promotes gp130-STAT3 signaling.

Adult-type intraembryonic hematopoiesis arises from specialized endothelial cells of the dorsal aorta (DA). Despite the critical importance of this specialized endothelium for establishment of hematopoietic stem cells and adult hematopoietic lineages, the mechanisms regulating its emergence are incompletely understood. We show that EphrinB2, a principal regulator of endothelial cell function, controls the development of endothelium producing adult-type hematopoiesis. The absence of EphrinB2 impairs DA-derived hematopoiesis. Transmembrane EphrinB2 and its EphB4 receptor interact in the emerging DA, which transiently harbors EphrinB2(+) and EphB4(+) endothelial cells, thereby providing an opportunity for bi-directional cell-to-cell signaling to control the emergence of the hemogenic endothelium. Embryonic Stem (ES) cell-derived EphrinB2(+) cells are enriched with hemogenic endothelial precursors. EphrinB2 silencing impairs ES generation of hematopoietic cells but not generation of endothelial cells. The identification of EphrinB2 as an essential regulator of adult hematopoiesis provides important insight in the regulation of early hematopoietic commitment.

EphrinB2 regulates the emergence of a hemogenic endothelium from the aorta.

Angiogenesis identifies the process of endothelial cell sprouting and remodeling leading to the formation of new and functional blood vessels. Vascular expansion during development and in the adult mammal provides nutrients and oxygen to areas with increased need. Although many molecules and pathways have been identified as regulators of angiogenesis, aspects of this complex process remain unclear. Particularly undefined are the signals that orchestrate vessel survival and pruning once new blood vessels have sprouted. These poorly characterized aspects of angiogenesis need exploration. This chapter describes the experiments and methods enabling the characterization of Semaphorin 6A as a critical regulator of endothelial cell survival and vessel function.

Characterization of Semaphorin 6A-Mediated Effects on Angiogenesis Through Regulation of VEGF Signaling.

Primary effusion lymphoma (PEL) is a Kaposi's sarcoma herpes virus (KSHV)-induced lymphoma that typically arises in body cavities of HIV-infected patients. PEL cells are often co-infected with Epstein-Barr virus (EBV). "PEL-like" lymphoma is a KSHV-unrelated lymphoma that arises in body cavities of HIV-negative patients. "PEL-like" lymphoma is sometimes EBV positive. The derivation of PEL/"PEL-like" cells is unclear.

Evidence for a Mesothelial Origin of Body Cavity Effusion Lymphomas.

Novel insights into endothelial cell malignancies.

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), has resulted in > 500,000 deaths worldwide, including > 125,000 deaths in the U.S. since its emergence in late December 2019 and June 2020. Neither curative anti-viral drugs nor a protective vaccine is currently available for the treatment and prevention of COVID-19. Recently, new clinical syndromes associated with coagulopathy and vasculopathy have emerged as a cause of sudden death and other serious clinical manifestations in younger patients infected with SARS-CoV-2 infection. Angiotensin converting enzyme 2 (ACE2), the receptor for SARS-CoV-2 and other coronaviruses, is a transmembrane protein expressed by lung alveolar epithelial cells, enterocytes, and vascular endothelial cells, whose physiologic role is to induce the maturation of angiotensin I to generate angiotensin 1-7, a peptide hormone that controls vasoconstriction and blood pressure. In this review, we provide the general context of the molecular and cellular mechanisms of SARS-CoV-2 infection with a focus on endothelial cells, describe the vasculopathy and coagulopathy syndromes in patients with SARS-CoV-2, and outline current understanding of the underlying mechanistic aspects.

Vasculopathy and Coagulopathy Associated with SARS-CoV-2 Infection.

Gene expression in individual cells is epigenetically regulated by DNA modifications, histone modifications, transcription factors, and other DNA-binding proteins. It has been shown that multiple histone modifications can predict gene expression and reflect future responses of bulk cells to extracellular cues. However, the predictive ability of epigenomic analysis is still limited for mechanistic research at a single cell level. To overcome this limitation, it would be useful to acquire reliable signals from multiple epigenetic marks in the same single cell. Here, we propose a new approach and a new method for analysis of several components of the epigenome in the same single cell. The new method allows reanalysis of the same single cell. We found that reanalysis of the same single cell is feasible, provides confirmation of the epigenetic signals, and allows application of statistical analysis to identify reproduced reads using data sets generated only from the single cell. Reanalysis of the same single cell is also useful to acquire multiple epigenetic marks from the same single cells. The method can acquire at least five epigenetic marks: H3K27ac, H3K27me3, mediator complex subunit 1, a DNA modification, and a DNA-interacting protein. We can predict active signaling pathways in K562 single cells using the epigenetic data and confirm that the predicted results strongly correlate with actual active signaling pathways identified by RNA-seq results. These results suggest that the new method provides mechanistic insights for cellular phenotypes through multilayered epigenome analysis in the same single cells.

Iterative epigenomic analyses in the same single cell.

The tyrosine phosphatase SHP2 is oncogenic in cancers driven by receptor-tyrosine-kinases, and SHP2 inhibition reduces tumor growth. Here, we report that SHP2 is an essential promoter of endothelial cell survival and growth in the remodeling tumor vasculature. Using genetic and chemical approaches to inhibit SHP2 activity in endothelial cells, we show that SHP2 inhibits pro-apoptotic STAT3 and stimulates proliferative ERK1/2 signaling. Systemic SHP2 inhibition in mice bearing tumor types selected for SHP2-independent tumor cell growth promotes degeneration of the tumor vasculature and blood extravasation; reduces tumor vascularity and blood perfusion; and increases tumor necrosis. Reduction of tumor growth ensues, independent of SHP2 targeting in the tumor cells, blocking immune checkpoints, or recruiting macrophages. We also show that inhibiting the Angiopoietin/TIE2/AKT cascade magnifies the vascular and anti-tumor effects of SHP2 inhibition by blocking tumor endothelial AKT signaling, not a target of SHP2. Since the SHP2 and Ang2/TIE2 pathways are active in vascular endothelial cells of human melanoma and colon carcinoma, SHP2 inhibitors alone or with Ang2/TIE2 inhibitors hold promise to effectively target the tumor endothelium.

Targeting the SHP2 phosphatase promotes vascular damage and inhibition of tumor growth.

Genetic and epigenetic lesions within hematopoietic cell populations drive diverse hematological malignancies. Myelodysplastic syndromes (MDS) are a group of myeloid neoplasms affecting the hematopoietic stem cells characterized by recurrent genetic abnormalities, myelodysplasia (a pathological definition of abnormal bone marrow structure), ineffective hematopoiesis resulting in blood cytopenia, and a propensity to evolve into acute myelogenous leukemia. Although there is evidence that the accumulation of a set of genetic mutations is an essential event in MDS, there is an increased appreciation of the contribution of specific microenvironments, niches, in the pathogenesis of MDS and response to treatment. In physiologic hematopoiesis, niches are critical functional units that maintain hematopoietic stem and progenitor cells and regulate their maturation into mature blood cells. In MDS and other hematological malignancies, altered bone marrow niches can promote the survival and expansion of mutant hematopoietic clones and provide a shield from therapy. In this review, we focus on our understanding of the composition and function of hematopoietic niches and their role in the evolution of myeloid malignancies, with an emphasis on MDS.

Hidetaka Ohnuki

Laboratory of Cellular Oncology,
Center for Cancer Research,
National Cancer Institute,
Laboratory of Cellular Oncology, Center for Cancer Research, National Cancer Institute

National Institutes of Health

Reusable Single Cell for Iterative Epigenomic Analyses

Hidetaka Ohnuki

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Laboratory of Cellular Oncology,Center for Cancer Research,National Cancer Institute,Laboratory of Cellular Oncology, Center for Cancer Research, National Cancer Institute

National Institutes of Health

Reusable Single Cell for Iterative Epigenomic Analyses

Laboratory of Cellular Oncology,
Center for Cancer Research,
National Cancer Institute,
Laboratory of Cellular Oncology, Center for Cancer Research, National Cancer Institute