bioprinting-cellularized-constructs-using-tissue-specific-hydrogel

Bioprinting Cellularized Constructs Using a Tissue-specific Hydrogel Bioink

Bioprinting has emerged as a versatile biofabrication approach for creating tissue engineered organ constructs. These constructs have potential use as ...

Wake Forest Univeristy Health Sciences

&#x87BA;&#x65CB;&#x87BA;&#x65CB;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x5BF9;&#x8BB8;&#x591A;&#x751F;&#x7269;&#x4FE1;&#x53F7;&#x4E0E;&#x7CFB;&#x7EDF;&#x81F3;&#x5173;&#x91CD;&#x8981;&#xFF0C;&#x5728;&#x76F4;&#x7ACB;&#x6216; heteromultimerization &#x7684;&#x4FE1;&#x53F7;&#x5206;&#x5B50;&#x4EE5;&#x53CA;&#x75C5;&#x6BD2;&#x8FDB;&#x5165;&#x5BBF;&#x4E3B;&#x4E2D;&#x627E;&#x5230;&#x3002;&#x5728;&#x827E;&#x6ECB;&#x75C5;&#x6BD2;&#xFF0C;&#x671F;&#x95F4;&#x611F;&#x67D3;&#x75C5;&#x6BD2;&#x4E0E;&#x5BBF;&#x4E3B;&#x819C;&#x878D;&#x5408;&#x88AB;&#x4ECB;&#x5BFC;&#x5F62;&#x6210;&#x7684;&#x516D;&#x4E2A;&#x87BA;&#x65CB;&#x675F; (6HBs) &#x4ECE; gp41&#xFF0C;homotrimers &#x6709;&#x75C5;&#x6BD2;&#x8FDB;&#x5165;&#x62EE;&#x6297;&#x5242;&#x4F5C;&#x4E3A;&#x6D3E;&#x751F;&#x7684;&#x5408;&#x6210;&#x80BD;&#x6570;&#x6240;&#x4ECE;&#x4E2D;&#x3002;&#x4F7F;&#x7528;&#x9175;&#x6BCD;&#x8868;&#x9762;&#x4E24;&#x4E2A;&#x6DF7;&#x5408; &#xFF08;YS2H&#xFF09; &#x7CFB;&#x7EDF;&#xFF0C;&#x4E00;&#x4E2A;&#x5E73;&#x53F0;&#xFF0C;&#x65E8;&#x5728;&#x68C0;&#x6D4B;&#x86CB;&#x767D;&#x8D28;-&#x86CB;&#x767D;&#x8D28;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x53D1;&#x751F;&#x901A;&#x8FC7;&#x5206;&#x6CCC;&#x7684;&#x9014;&#x5F84;&#xFF0C;&#x6211;&#x4EEC;&#x91CD;&#x7EC4;&#x9175;&#x6BCD;&#x8868;&#x9762;&#x4E0A;&#x770B;&#xFF0C;6HB &#x914D;&#x5408;&#x7269;&#x5B9A;&#x91CF;&#x8861;&#x91CF;&#x7684;&#x5E73;&#x8861;&#x548C;&#x52A8;&#x529B;&#x5B66;&#x5E38;&#x6570;&#x7684;&#x53EF;&#x6EB6;&#x6027; 6HB&#xFF0C;&#x548C;&#x5212;&#x5B9A;&#x7684;&#x6B8B;&#x7559;&#x5F71;&#x54CD;&#x540C;&#x6E90;&#x5BE1;&#x805A;&#x548C;&#x5F02;&#x8D28;&#x5BE1;&#x805A;&#x5377;&#x7EBF;&#x5708;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3002;&#x56E0;&#x6B64;&#xFF0C;&#x6211;&#x4EEC;&#x76EE;&#x524D; YS2H &#x4F5C;&#x4E3A;&#x4E00;&#x4E2A;&#x5E73;&#x53F0;&#x7684;&#x7B80;&#x6613;&#x8868;&#x5F81;&#x4E0E;&#x827E;&#x6ECB;&#x75C5;&#x6BD2;&#x548C;&#x4F9D;&#x9760;&#x819C;&#x878D;&#x5408;&#x7684;&#x611F;&#x67D3;&#xFF0C;&#x4EE5;&#x53CA;&#x89E6;&#x53D1;&#x7684;&#x5F02;&#x8D28;&#x5BE1;&#x805A;&#x5377;&#x66F2;&#x7684;&#x8702;&#x7A9D;&#x4FE1;&#x53F7;&#x4E8B;&#x4EF6;&#x7684;&#x8BB8;&#x591A;&#x5176;&#x4ED6;&#x5305;&#x819C;&#x7684;&#x75C5;&#x6BD2;&#x7684;&#x6291;&#x5236;&#x4F5C;&#x7528;&#x62EE;&#x6297;&#x591A;&#x80BD;&#x7684;&#x8BBE;&#x8BA1;&#x3002;

&#x5355;&#x5143;&#x683C;&#x7684;&#x827E;&#x6ECB;&#x75C5;&#x6BD2;&#x8868;&#x9762;&#x5927;&#x4F1A; gp41 &#x516D;&#x87BA;&#x65CB;&#x675F;&#x4E3A; Facile&#xFF0C;&#x5F02;&#x8D28;&#x5BE1;&#x805A;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x7684;&#x5B9A;&#x91CF;&#x6D4B;&#x91CF;&#x7684;&#x3002;

Helix-helix interactions are fundamental to many biological signals and systems and are found in homo- or heteromultimerization of signaling molecules as well as in the process of virus entry into the host. In HIV, virus-host membrane fusion during infection is mediated by the formation of six-helix bundles (6HBs) from homotrimers of gp41, from which a number of synthetic peptides have been derived as antagonists of virus entry. Using a yeast surface two-hybrid (YS2H) system, a platform designed to detect protein-protein interactions occurring through a secretory pathway, we reconstituted 6HB complexes on the yeast surface, quantitatively measured the equilibrium and kinetic constants of soluble 6HB, and delineated the residues influencing homo-oligomeric and hetero-oligomeric coiled-coil interactions. Hence, we present YS2H as a platform for the facile characterization and design of antagonistic peptides for inhibition of HIV and many other enveloped viruses relying on membrane fusion for infection, as well as cellular signaling events triggered by hetero-oligomeric coiled coils.

Cell Surface Assembly of HIV gp41 Six-Helix Bundles for Facile, Quantitative Measurements of Hetero-oligomeric Interactions.

H&eacute;lice-h&eacute;lice interactions sont fondamentales pour de nombreux syst&egrave;mes et signaux biologiques et sont retrouvent dans homo - ou heteromultimerization des mol&eacute;cules de signalisation, ainsi que dans le processus d&#39;entr&eacute;e du virus dans l&#39;h&ocirc;te. En mati&egrave;re de VIH, fusion membranaire virus-h&ocirc;te au cours de l&#39;infection est favoris&eacute;e par la formation de faisceaux de six-h&eacute;lice (6HBs) de homotrimers de la gp41, d&#39;o&ugrave; un certain nombre de peptides synth&eacute;tiques ont &eacute;t&eacute; d&eacute;riv&eacute; en tant qu&#39;antagonistes de l&#39;entr&eacute;e du virus. &Agrave; l&#39;aide d&#39;un surface deux syst&egrave;me hybride de levure (YS2H), une plateforme con&ccedil;ue pour d&eacute;tecter des interactions prot&eacute;ine-prot&eacute;ine, dus &agrave; une voie de s&eacute;cr&eacute;tion, que nous avons reconstitu&eacute; 6HB complexes sur la surface de la levure, mesur&eacute;e quantitativement l&#39;&eacute;quilibre et les constantes cin&eacute;tiques de 6HB soluble et d&eacute;limit&eacute; les r&eacute;sidus influen&ccedil;ant homo-oligom&egrave;res et h&eacute;t&eacute;ro-oligom&eacute;riques interactions coiled-coil. C&#39;est pourquoi nous pr&eacute;sentons YS2H comme une plate-forme pour la caract&eacute;risation facile et la conception des peptides antagonistes pour l&#39;inhibition du VIH et de nombreux autres virus envelopp&eacute;s en s&#39;appuyant sur la fusion membranaire pour l&#39;infection, ainsi que des &eacute;v&eacute;nements de signalisation cellulaires d&eacute;clench&eacute;s par des bobines d&#39;enroulement h&eacute;t&eacute;ro-oligom&egrave;res.

Cell Surface Assembl&eacute;e du VIH gp41 Six-Helix Bundles pour Facile, des mesures quantitatives des Interactions h&eacute;t&eacute;ro-oligom&egrave;res.

Helix-Helix-Wechselwirkungen sind sind grundlegend f&uuml;r viele biologische Signale und Systeme und in Homo- oder Heteromultimerization signalisieren der Molek&uuml;le sowie im Virus-Eintrag in der Host-Prozess. HIV ist Virus-Host Membrane Fusion w&auml;hrend der Infektion durch die Bildung von sechs-Helix-B&uuml;ndel (6HBs) vermittelte aus Homotrimers von gp41, von der haben eine Reihe von synthetischen Peptide als Antagonisten f&uuml;r Virusangriffe abgeleitet wurde. Mit einem Hefe Oberfl&auml;che zwei-Mischling (YS2H) System, eine Plattform f&uuml;r das Aufsp&uuml;ren von Protein-Protein-Interaktionen, die durch einen sekretorischen Weg, wir 6HB-komplexe an der Oberfl&auml;che Hefe rekonstituiert quantitativ gemessen, das Gleichgewicht und die kinetischen Konstanten der l&ouml;slichen 6HB und abgegrenzt, die R&uuml;ckst&auml;nde, die Beeinflussung der Homo-Oligomere und Hetero-Oligomere aufgerollte Spule-Interaktionen. Daher stellen wir YS2H als Plattform f&uuml;r die oberfl&auml;chlichen Charakterisierung und Gestaltung der antagonistischen Peptide f&uuml;r die Hemmung von HIV und viele andere umh&uuml;llten Viren unter Berufung auf Membrane Fusion f&uuml;r Infektion sowie zellulare signalisierende Ereignisse, die ausgel&ouml;st durch Hetero-Oligomere gewickelten Spulen.

Sechs-Helix-Bundles von Zelle Oberfl&auml;che Montage von HIV gp41 Facile, Quantitative Messungen der Hetero-Oligomere Interaktionen.

Elica-elica interazioni fondamentali per molti segnali biologici e sistemi e si trovano in homo - o heteromultimerization di molecole di segnalazione, cos&igrave; come nel processo di entrata del virus nell&#39;ospite. In HIV, fusione membrana virus-ospite durante l&#39;infezione &egrave; mediato dalla formazione di sei-helix Bundle (6HBs) da homotrimers di gp41, da cui un certo numero di peptidi sintetici derivato come antagonisti di entrata del virus. Utilizzando un superficie due-sistema ibrido del lievito (YS2H), una piattaforma progettata per rilevare le interazioni della proteina-proteina, che si verificano attraverso un pathway secretiva, che abbiamo ricostituito 6HB complessi sulla superficie del lievito, misurarono quantitativamente l&#39;equilibrio e le costanti cinetiche di 6HB solubile e delineati i residui che influenzano homo-oligomerica ed etero-oligomerica interazioni coiled-coil. Quindi, vi presentiamo YS2H come una piattaforma per la facile caratterizzazione e progettazione di peptidi antagonisti per inibizione dell&#39;HIV e molti altri virus avvolto basandosi sulla fusione di membrana per infezione, cos&igrave; come eventi di segnalazione cellulari innescati da etero-oligomerica bobine arrotolati.

Cella superficie Assembly of HIV gp41 sei-Helix bundle per Facile, le misure Quantitative di etero-oligomerica interazioni.

&#x30D8;&#x30EA;&#x30C3;&#x30AF;&#x30B9; &#x30D8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x9593;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306F;&#x3001;&#x591A;&#x304F;&#x306E;&#x751F;&#x4F53;&#x4FE1;&#x53F7;&#x3068;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306B;&#x57FA;&#x672C;&#x3067;&#x3042;&#x308A;&#x30DB;&#x30E2;&#x307E;&#x305F;&#x306F; heteromultimerization &#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x5206;&#x5B50;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x3001;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306E;&#x30A8;&#x30F3;&#x30C8;&#x30EA;&#x306B;&#x30DB;&#x30B9;&#x30C8;&#x306E;&#x904E;&#x7A0B;&#x3067;&#x767A;&#x898B;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;HIV &#x306E;&#x30A6;&#x30A4;&#x30EB;&#x30B9;-&#x5BBF;&#x4E3B;&#x819C;&#x878D;&#x5408;&#x611F;&#x67D3;&#x6642;&#x306B; 6 &#x30D8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x30D0;&#x30F3;&#x30C9;&#x30EB; (6HBs) &#x306E;&#x5F62;&#x6210;&#x306B;&#x3088;&#x3063;&#x3066;&#x3092;&#x304B;&#x3089;&#x30A6;&#x30A4;&#x30EB;&#x30B9; &#x30A8;&#x30F3;&#x30C8;&#x30EA;&#x306E;&#x62EE;&#x6297;&#x85AC;&#x3068;&#x3057;&#x3066;&#x591A;&#x6570;&#x306E;&#x5408;&#x6210;&#x30DA;&#x30D7;&#x30C1;&#x30C9;&#x3092;&#x6D3E;&#x751F;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B; gp41 &#x306E; homotrimers &#x304B;&#x3089;&#x4EF2;&#x4ECB;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x9175;&#x6BCD;&#x8868;&#x9762; 2 &#x30CF;&#x30A4;&#x30D6;&#x30EA;&#x30C3;&#x30C9; (YS2H) &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3001;&#x6211;&#x3005; 6HB &#x932F;&#x4F53;&#x9175;&#x6BCD;&#x8868;&#x9762;&#x518D;&#x69CB;&#x6210;&#x306F;&#x5206;&#x6CCC;&#x7D4C;&#x8DEF;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x767A;&#x751F;&#x3057;&#x3066;&#x86CB;&#x767D;&#x8CEA;&#x86CB;&#x767D;&#x8CEA;&#x306E;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3092;&#x691C;&#x51FA;&#x3059;&#x308B;&#x3088;&#x3046;&#x306B;&#x8A2D;&#x8A08;&#x30D7;&#x30E9;&#x30C3;&#x30C8;&#x30D5;&#x30A9;&#x30FC;&#x30E0;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x5E73;&#x8861;&#x3068;&#x53CD;&#x5FDC;&#x306B;&#x304A;&#x3051;&#x308B;&#x901F;&#x5EA6;&#x5B9A;&#x6570;&#x53EF;&#x6EB6;&#x6027; 6HB &#x306E;&#x5B9A;&#x91CF;&#x7684;&#x6E2C;&#x5B9A;&#x3057;&#x3001;&#x30DB;&#x30E2; &#x30AA;&#x30EA;&#x30B4;&#x30DE;&#x30FC;&#x306B;&#x5F71;&#x97FF;&#x3092;&#x53CA;&#x307C;&#x3059;&#x6B8B;&#x57FA;&#x3068;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x30B3;&#x30A4;&#x30EB;&#x30C9; &#x30B3;&#x30A4;&#x30EB; &#x30D8;&#x30C6;&#x30ED; &#x30AA;&#x30EA;&#x30B4;&#x30DE;&#x30FC;&#x7DDA;&#x5F15;&#x304D;&#x3002;&#x305D;&#x308C;&#x6545;&#x306B;&#x3001;&#x5B89;&#x6613;&#x306A;&#x30AD;&#x30E3;&#x30E9;&#x30AF;&#x30BF;&#x30EA;&#x30BC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3068;&#x62EE;&#x6297;&#x30DA;&#x30D7;&#x30C1;&#x30C9;&#x306E; HIV &#x30FB;&#x819C;&#x878D;&#x5408;&#x30D8;&#x30C6;&#x30ED; &#x30AA;&#x30EA;&#x30B4;&#x30DE;&#x30FC;&#x306E;&#x30B3;&#x30A4;&#x30EB;&#x30C9; &#x30B3;&#x30A4;&#x30EB;&#x306B;&#x3088;&#x3063;&#x3066;&#x30C8;&#x30EA;&#x30AC;&#x30FC;&#x3055;&#x308C;&#x308B;&#x7D30;&#x80DE;&#x306E;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x30A4;&#x30D9;&#x30F3;&#x30C8;&#x3068;&#x540C;&#x69D8;&#x306B;&#x611F;&#x67D3;&#x75C7;&#x3001;&#x305F;&#x3081;&#x306B;&#x983C;&#x308B;&#x591A;&#x304F;&#x306E;&#x4ED6;&#x306E;&#x30A8;&#x30F3;&#x30D9;&#x30ED;&#x30FC;&#x30D7; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x6291;&#x5236;&#x306E;&#x305F;&#x3081;&#x306E;&#x8A2D;&#x8A08;&#x306E;&#x305F;&#x3081;&#x306E;&#x30D7;&#x30E9;&#x30C3;&#x30C8;&#x30D5;&#x30A9;&#x30FC;&#x30E0;&#x3068;&#x3057;&#x3066; YS2H &#x3092;&#x63D0;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

&#x8868;&#x9762; HIV &#x30A2;&#x30BB;&#x30F3;&#x30D6;&#x30EA; gp41 6 &#x30D8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x30D0;&#x30F3;&#x30C9;&#x30EB; &#x30D8;&#x30C6;&#x30ED; &#x30AA;&#x30EA;&#x30B4;&#x30DE;&#x30FC;&#x306E;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x5B9A;&#x91CF;&#x7684;&#x6E2C;&#x5B9A;&#x30D5;&#x30A1;&#x30B7;&#x30EB;&#x306E;&#x30BB;&#x30EB;&#x3057;&#x307E;&#x3059;&#x3002;

&#xD5EC;&#xB9AD;&#xC2A4; &#xB098;&#xC120; &#xC0C1;&#xD638; &#xC791;&#xC6A9; &#xB9CE;&#xC740; &#xC0DD;&#xBB3C; &#xD559;&#xC801; &#xC2E0;&#xD638; &#xBC0F; &#xC2DC;&#xC2A4;&#xD15C; &#xAE30;&#xBCF8; &#xC774;&#xBA70; &#xD638;&#xBAA8;-&#xB610;&#xB294; &#xD638;&#xC2A4;&#xD2B8;&#xC5D0; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xD56D;&#xBAA9; &#xACFC;&#xC815; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; &#xC2E0;&#xD638; &#xBD84;&#xC790;&#xC758; heteromultimerization&#xC5D0;&#xC11C; &#xCC3E;&#xC744; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5D0;&#xC774;&#xC988;, &#xAC10;&#xC5FC; &#xC2DC; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xD638;&#xC2A4;&#xD2B8; &#xB9C9; &#xD4E8;&#xC804; &#xC911;&#xC7AC; 6 &#xB098;&#xC120; &#xBC88;&#xB4E4; (6HBs)&#xC758; &#xD615;&#xC131;&#xC5D0; &#xC758;&#xD574; gp41, homotrimers&#xC5D0;&#xC11C;&#xB294; &#xB2E4;&#xC591; &#xD55C; &#xD569;&#xC131; &#xD3A9; &#xD2F0; &#xB4DC;&#xB294; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xD56D;&#xBAA9;&#xC758; &#xAE38; &#xD56D; &#xADFC;&#xC73C;&#xB85C; &#xD30C;&#xC0DD; &#xB41C; &#xAC83;&#xC785;&#xB2C8;&#xB2E4;. &#xD6A8; &#xBAA8; &#xD45C;&#xBA74; 2-&#xD558;&#xC774;&#xBE0C;&#xB9AC;&#xB4DC; (YS2H) &#xC2DC;&#xC2A4;&#xD15C;, &#xC6B0;&#xB9AC; &#xD6A8; &#xBAA8; &#xD45C;&#xBA74;&#xC5D0; 6HB &#xB2E8;&#xC9C0; &#xC7AC;&#xAD6C;&#xC131; &#xBD84; &#xD1B5;&#xB85C; &#xD1B5;&#xD574; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xB2E8;&#xBC31;&#xC9C8; &#xB2E8;&#xBC31;&#xC9C8; &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC744; &#xAC10;&#xC9C0; &#xD558;&#xB3C4;&#xB85D; &#xC124;&#xACC4; &#xB41C; &#xD50C;&#xB7AB;&#xD3FC;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC591;&#xC801; &#xADE0;&#xD615; &#xBC0F; &#xC6A9; &#xD574; 6hb&#xC758; &#xD0A4;&#xB124;&#xD2F1; &#xC0C1;&#xC218;&#xB97C; &#xCE21;&#xC815; &#xD558; &#xACE0; &#xD638;&#xBAA8; oligomeric&#xC5D0; &#xC601;&#xD5A5;&#xC744; &#xBBF8;&#xCE58;&#xB294; &#xC794;&#xB958;&#xBB3C; &#xBC0F; &#xC774;&#xC885; oligomeric &#xCF54;&#xC77C; &#xCF54;&#xC77C; &#xC0C1;&#xD638; &#xC791;&#xC6A9; delineated. &#xB530;&#xB77C;&#xC11C;, &#xC120;&#xBB3C;&#xC774; &#xC190;&#xC26C;&#xC6B4; &#xD2B9;&#xC131;&#xD654; &#xBC0F; HIV&#xC640; hetero oligomeric &#xCF54;&#xC77C;&#xB41C; &#xCF54;&#xC77C;&#xC5D0; &#xC758;&#xD574; &#xBC1C;&#xC0DD; &#xB41C; &#xC140;&#xB8F0;&#xB7EC; &#xC2E0;&#xD638; &#xC774;&#xBCA4;&#xD2B8; &#xBC0F; &#xAC10;&#xC5FC;&#xC5D0; &#xB300; &#xD55C; &#xB9C9; &#xC735;&#xD569;&#xC5D0; &#xC758;&#xC874; &#xD558;&#xB294; &#xB2E4;&#xB978; &#xB9CE;&#xC740; &#xC5D4;&#xBC8C;&#xB85C;&#xD504;&#xB41C; &#xBC14;&#xC774;&#xB7EC;&#xC2A4;&#xC758; &#xC5B5;&#xC81C;&#xC5D0; &#xB300; &#xD55C; &#xB300;&#xB9BD; &#xD3A9; &#xD2F0; &#xB4DC;&#xC758; &#xB514;&#xC790;&#xC778;&#xC744; &#xC704;&#xD55C; &#xD50C;&#xB7AB;&#xD3FC;&#xC73C;&#xB85C; YS2H.

&#xC140; &#xD45C;&#xBA74; &#xC5B4;&#xC148;&#xBE14;&#xB9AC; &#xC5D0;&#xC774;&#xC988; gp41 6 &#xB098;&#xC120; &#xBC88;&#xB4E4; Facile, Hetero oligomeric &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC758; &#xC815;&#xB7C9;&#xC801; &#xCE21;&#xC815;&#xC5D0; &#xB300; &#xD55C;.

H&eacute;lice h&eacute;lice interacciones son fundamentales para muchas se&ntilde;ales biol&oacute;gicas y sistemas y se encuentran en Homo o heteromultimerization de las mol&eacute;culas de se&ntilde;alizaci&oacute;n, as&iacute; como en el proceso de entrada de virus en el host. En VIH, fusi&oacute;n de membrana virus-hu&eacute;sped durante la infecci&oacute;n est&aacute; mediada por la formaci&oacute;n de la h&eacute;lice seis paquetes (6HBs) de homotrimers de gp41, de que un n&uacute;mero de p&eacute;ptidos sint&eacute;ticos derivado como antagonistas de entrada de virus. Utilizando un levadura superficial (YS2H) sistema del dos-h&iacute;brido, una plataforma dise&ntilde;ada para detectar interacciones prote&iacute;na-prote&iacute;na que ocurre a trav&eacute;s de una v&iacute;a secretora, que hemos reconstituido 6HB complejos en la superficie de la levadura, medici&oacute;n cuantitativamente el equilibrio y constantes cin&eacute;ticas de 6HB solubles y deline&oacute; los residuos que influyen en los homo y hetero-oligom&eacute;ricas interacciones coiled-coil. Por lo tanto, presentamos YS2H como una plataforma para la f&aacute;cil caracterizaci&oacute;n y dise&ntilde;o de p&eacute;ptidos antag&oacute;nicos para la inhibici&oacute;n del VIH y muchos otros virus envueltos, confiando en la fusi&oacute;n de la membrana para la infecci&oacute;n, as&iacute; como eventos de se&ntilde;alizaci&oacute;n celulares activados por hetero-oligom&eacute;ricas bobinas en espiral.

Celular montaje de superficie del VIH gp41 h&eacute;lice seis paquetes para Facile, mediciones cuantitativas de interacciones Hetero-oligom&eacute;ricas.

Current in vitro models for tumor growth and metastasis are poor facsimiles of in vivo cancer physiology and thus, are not optimal for anti-cancer drug development. Three dimensional (3D) tissue organoid systems, which utilize human cells in a tailored microenvironment, have the potential to recapitulate in vivo conditions and address the drawbacks of current tissue culture dish 2D models. In this study, we created liver-based cell organoids in a rotating wall vessel bioreactor. The organoids were further inoculated with colon carcinoma cells in order to create liver-tumor organoids for in vitro modeling of liver metastasis. Immunofluorescent staining revealed notable phenotypic differences between tumor cells in 2D and inside the organoids. In 2D they displayed an epithelial phenotype, and only after transition to the organoids did the cells present with a mesenchymal phenotype. The cell surface marker expression results suggested that WNT pathway might be involved in the phenotypic changes observed between cells in 2D and organoid conditions, and may lead to changes in cell proliferation. Manipulating the WNT pathway with an agonist and antagonist showed significant changes in sensitivity to the anti-proliferative drug 5-fluoruracil. Collectively, the results show the potential of in vitro 3D liver-tumor organoids to serve as a model for metastasis growth and for testing the response of tumor cells to current and newly discovered drugs.

Liver-Tumor Hybrid Organoids for Modeling Tumor Growth and Drug Response In Vitro.

Advancement of bioprinting technology is limited by the availability of materials that both facilitate bioprinting logistics as well as support cell viability and function by providing tissue-specific cues. Herein we describe a modular hyaluronic acid (HA) and gelatin-based hydrogel toolbox comprised of a 2-crosslinker, 2-stage polymerization technique, and the capability to provide tissue specific biochemically and mechanically accurate signals to cells within biofabricated tissue constructs. First, we prepared and characterized several tissue-derived decellularized extracellular matrix-based solutions, which contain complex combinations of growth factors, collagens, glycosaminoglycans, and elastin. These solutions can be incorporated into bioinks to provide the important biochemical cues of different tissue types. Second, we employed combinations of PEG-based crosslinkers with varying molecular weights, geometries (linear, 4-arm, and 8-arm), and functional groups to yield hydrogel bioinks that supported extrusion bioprinting and the capability to achieve final construct shear stiffness values ranging from approximately 100 Pa to 20 kPa. Lastly, we integrated these hydrogel bioinks with a 3-D bioprinting platform, and validated their use by bioprinting primary liver spheroids in a liver-specific bioink to create in vitro liver constructs with high cell viability and measurable functional albumin and urea output. This hydrogel bioink system has the potential to be a versatile tool for biofabrication of a wide range of tissue construct types.

A hydrogel bioink toolkit for mimicking native tissue biochemical and mechanical properties in bioprinted tissue constructs.

Current animal and 2-D cell culture models employed in metastasis research and drug discovery remain poor mimics of human cancer physiology. Here we describe a "metastasis-on-a-chip" system allowing real time tracking of fluorescent colon cancer cells migrating from hydrogel-fabricated gut constructs to downstream liver constructs within a circulatory fluidic device system that responds to environmental manipulation and drug treatment. Devices consist of two chambers in which gut and liver constructs are housed independently, but are connected in series via circulating fluid flow. Constructs were biofabricated with a hyaluronic acid-based hydrogel system, capable of a variety of customizations, inside of which representative host tissue cells were suspended and metastatic colon carcinoma tumor foci were created. The host tissue of the constructs expressed normal epithelial markers, which the tumor foci failed to express. Instead, tumor regions lost membrane-bound adhesion markers, and expressed mesenchymal and proliferative markers, suggesting a metastatic phenotype. Metastatic tumor foci grew in size, eventually disseminating from the intestine construct and entering circulation, subsequently reaching in the liver construct, thus mimicking some of the migratory events observed during metastasis. Lastly, we demonstrated the ability to manipulate the system, including chemically modulating the hydrogel system mechanical properties and administering chemotherapeutic agents, and evaluated the effects of these parameters on invasive tumor migration. These results describe the capability of this early stage metastasis-on-a-chip system to model several important characteristics of human metastasis, thereby demonstrating the potential of the platform for making meaningful advances in cancer investigation and drug discovery. This article is protected by copyright. All rights reserved.

Article	Year
Cell Surface Assembly of HIV gp41 Six-Helix Bundles for Facile, Quantitative Measurements of Hetero-oligomeric Interactions. Journal of the American Chemical Society\| PubMed ID: 22888993	2012
Liver-Tumor Hybrid Organoids for Modeling Tumor Growth and Drug Response In Vitro. Annals of biomedical engineering\| PubMed ID: 25777294	2015
A hydrogel bioink toolkit for mimicking native tissue biochemical and mechanical properties in bioprinted tissue constructs. Acta biomaterialia\| PubMed ID: 26210285	2015
A reductionist metastasis-on-a-chip platform for in vitro tumor progression modeling and drug screening. Biotechnology and bioengineering\| PubMed ID: 26888480	2016

Mahesh Devarasetty

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