Mouse Models for Graft Arteriosclerosis

Graft arteriosclerois (GA),  also called allograft vasculopathy, is a pathologic lesion that develops over  months to years in transplanted organs ...

Yale University School of Medicine 

Xenotransplantation from pig to human being is viewed as a potential solution for the acute organ shortage. However, consequent xenorejection induced by Gal alpha 1,3 Gal (Gal, Gal antigen) prevents xenotransplantation from clinical application. Thus, the most attracting attempt to prevent xenorejection is the elimination of Gal. Our study suggested that compared with the human alpha 1,2 fucosyltransferase (FT) gene and porcine antisense alpha 1,3 galactosyltransferase gene, sequence-specific siRNA targeting Gal were capable of suppressing Gal expression markedly, and therefore, significantly inhibiting xenoreactivity and the complement activation with human serum in PIEC cells. We also demonstrated the concordant inhibitory effect of siRNA and human FT gene on Gal and corresponding functions, which implied a practical significance of combined transgenic strategy. The successful application of vector-based dsRNA-GT may extend the list of available modalities in the abrogation of xenorejection in xenotransplantation.

Effect of RNA interference on Gal alpha 1,3 Gal expression in PIEC cells.

X&eacute;notransplantation du cochon &agrave; l&#39;&ecirc;tre humain est consid&eacute;r&eacute;e comme une solution potentielle &agrave; la p&eacute;nurie d&#39;organe aigu&euml;. Cependant, xenorejection cons&eacute;quente induite par le Gal alpha 1,3 liste d&#39;adresses globale (Gal, Gal antig&egrave;ne) emp&ecirc;che la x&eacute;notransplantation d&#39;application clinique. Ainsi, la tentative plus attire pour emp&ecirc;cher xenorejection est l&#39;&eacute;limination de la liste d&#39;adresses globale. Notre &eacute;tude a sugg&eacute;r&eacute; que par rapport au g&egrave;ne de fucosyltransf&eacute;rase (FT) alpha humain 1,2 et g&egrave;ne porcin antisens alpha 1,3 galactosyltransf&eacute;rase s&eacute;quence sp&eacute;cifique siARN ciblant les Gal &eacute;taient capables de suppression de Gal expression nettement et, par cons&eacute;quent, sensiblement inhibant la xenoreactivity et l&#39;activation du compl&eacute;ment avec le s&eacute;rum humain dans les cellules PIEC. Nous avons aussi d&eacute;montr&eacute; l&#39;effet inhibiteur concordante des siARN et le g&egrave;ne humain de FT sur la liste d&#39;adresses globale et les fonctions correspondantes, qui impliquait une importance concr&egrave;te de cette strat&eacute;gie combin&eacute;e de transg&eacute;nique. L&#39;application r&eacute;ussie de dsRNA-GT vectoriel peut &eacute;tendre la liste des modalit&eacute;s disponibles dans l&#39;abrogation de xenorejection dans la x&eacute;notransplantation.

Effet de l&#39;interf&eacute;rence d&#39;ARN sur Gal alpha 1,3 Gal expression dans les cellules PIEC.

Lo xenotrapianto da maiale all&#39;essere umano &egrave; visto come una potenziale soluzione per la carenza di organo acuta. Tuttavia, la conseguente xenorejection indotta da Gal alfa 1,3 elenco indirizzi globale (Gal, Gal antigene) impedisce lo xenotrapianto da applicazione clinica. Cos&igrave;, il tentativo di attirare pi&ugrave; per evitare che xenorejection &egrave; l&#39;eliminazione del Gal. Il nostro studio ha suggerito che confrontato con il gene fucosiltransferasi (FT) Alfa 1,2 e gene galattosiltransferasi porcine antisenso alfa 1,3 sequenza-specifici siRNA targeting Gal sono stati in grado di sopprimere il Gal espressione marcatamente e pertanto, inibendo significativamente xenoreactivity e attivazione del complemento con siero umano in cellule PIEC. Abbiamo anche dimostrato l&#39;effetto inibitorio concordante di siRNA e gene umano FT su Gal e le funzioni corrispondenti, che implicava un significato pratico della strategia combinata transgenico. L&#39;applicazione di successo di dsRNA-GT vettoriale possa estendere l&#39;elenco delle modalit&agrave; disponibili in abrogazione del xenorejection in xenotrapianti.

Effetto di interferenza del RNA su Gal alfa 1,3 Gal espressione in cellule PIEC.

Xenotransplantation from pigs to human beings is viewed as a potential solution for the acute organ shortage. However, consequent xenorejection induced by Gal alpha 1,3 Gal (a Gal, Gal antigen) prevents xenotransplantation from clinical application. Thus, the most attracting attempt to prevent xenorejection is the elimination of Gal. Our study suggested that compared with the human alpha 1,2 fucosyltransferase (FT) gene and the porcine antisense alpha 1,3 galactosyltransferase gene, sequence-specific siRNA targeting Gal was capable of suppressing Gal expression markedly, and therefore, significantly inhibiting xenoreactivity and the complement activation with human serum in PIEC cells. We also demonstrated the concordant inhibitory effect of siRNA and the human FT gene on Gal and corresponding functions, which implied a practical significance of combined transgenic strategy. The successful application of vector-based dsRNA-GT may extend the list of available modalities in the abrogation of xenorejection in xenotransplantation.

X&eacute;notransplantation provenant de porcs &agrave; l&#39;&ecirc;tre humain est consid&eacute;r&eacute;e comme une solution potentielle &agrave; la p&eacute;nurie d&#39;organe aigu&euml;. Cependant, xenorejection cons&eacute;quente induite par le Gal alpha 1,3 Gal (un Gal, Gal antig&egrave;ne) emp&ecirc;che la x&eacute;notransplantation d&#39;application clinique. Ainsi, la tentative plus attire pour emp&ecirc;cher xenorejection est l&#39;&eacute;limination de la liste d&#39;adresses globale. Notre &eacute;tude a sugg&eacute;r&eacute; que par rapport au g&egrave;ne de fucosyltransf&eacute;rase (FT) humaine alpha 1, 2, et le g&egrave;ne de la galactosyltransf&eacute;rase porcine antisens alpha 1, 3, s&eacute;quence sp&eacute;cifique siARN ciblant Gal &eacute;tait capable de suppression de Gal expression nettement et, par cons&eacute;quent, sensiblement inhibant la xenoreactivity et l&#39;activation du compl&eacute;ment avec le s&eacute;rum humain dans les cellules PIEC. Nous avons aussi d&eacute;montr&eacute; l&#39;effet inhibiteur concordante des siARN et le g&egrave;ne humain de FT sur la liste d&#39;adresses globale et les fonctions correspondantes, qui impliquait une importance concr&egrave;te de cette strat&eacute;gie combin&eacute;e de transg&eacute;nique. L&#39;application r&eacute;ussie de dsRNA-GT vectoriel peut &eacute;tendre la liste des modalit&eacute;s disponibles dans l&#39;abrogation de xenorejection dans la x&eacute;notransplantation.

Lo xenotrapianto da maiali agli esseri umani &egrave; visto come una potenziale soluzione per la carenza di organo acuta. Tuttavia, la conseguente xenorejection indotta da Gal alfa 1,3 Gal (un Gal, Gal antigene) impedisce lo xenotrapianto da applicazione clinica. Cos&igrave;, il tentativo di attirare pi&ugrave; per evitare che xenorejection &egrave; l&#39;eliminazione del Gal. Il nostro studio ha suggerito che rispetto al gene umano alfa 1,2 fucosiltransferasi (FT) e il gene galattosiltransferasi porcine antisenso alfa 1,3, sequenza-specifici siRNA targeting Gal &egrave; stato in grado di sopprimere il Gal espressione marcatamente e pertanto, inibendo significativamente xenoreactivity e attivazione del complemento con siero umano in cellule PIEC. Abbiamo anche dimostrato l&#39;effetto inibitorio concordante di siRNA e il gene umano FT su Gal e le funzioni corrispondenti, che implicava un significato pratico della strategia combinata transgenico. L&#39;applicazione di successo di dsRNA-GT vettoriale possa estendere l&#39;elenco delle modalit&agrave; disponibili in abrogazione del xenorejection in xenotrapianti.

Focal cerebral ischemia causes apoptosis in neural cells during the postischemia period. TNF is critically involved in such neuronal apoptosis mediated by caspase pathways. A20 can inhibit TNF-induced apoptosis in many cell types. However, little work has been carried out in central nervous system. In the present study, gene transfer of A20 resulted in reduction of infarct volume and improvement of neurological deficit in ischemia rats. Results of flow cytometry, TUNEL and DNA fragmentation assay all indicated A20 could inhibit TNF-induced apoptosis both in primary rat hippocampal neurons and SH-SY5Y cells. Moreover, we found A20 targeted the TNF apoptotic pathway by inhibiting proteolytic cleavage of caspase 8 and 3 in SH-SY5Y cells. These data demonstrated A20 could effectively protect neurons from postischemic apoptosis and may function partly on death receptor caspase pathway. Gene transfer of A20 may be a promising approach to gene therapy for cerebral ischemia in the future.

Neuroprotective effect of A20 on TNF-induced postischemic apoptosis.

L&#39;isch&eacute;mie c&eacute;r&eacute;brale provoque l&#39;apoptose dans les cellules neurales au cours de la p&eacute;riode postischemia. TNF est critique impliqu&eacute; dans telle l&#39;apoptose neuronale m&eacute;di&eacute;e par des voies de caspase. A20 peut inhiber l&#39;apoptose induite par le TNF dans de nombreux types de cellules. Cependant, peu de travaux a &eacute;t&eacute; effectu&eacute;e dans le syst&egrave;me nerveux central. Dans la pr&eacute;sente &eacute;tude, transfert de g&egrave;nes de l&#39;A20 a entra&icirc;n&eacute; dans la r&eacute;duction du volume de l&#39;infarctus et l&#39;am&eacute;lioration du d&eacute;ficit neurologique chez les rats de l&#39;isch&eacute;mie. R&eacute;sultats de cytom&eacute;trie en flux, essai de fragmentation de TUNEL et ADN tous indiqu&eacute; qu&#39;a20 capables d&#39;inhiber l&#39;apoptose induite par le TNF en neurones hippocampiques primaires de rat et de cellules SH-SY5Y. En outre, nous avons trouv&eacute; qu&#39;a20 cibl&eacute; la voie apoptotique TNF en inhibant le clivage prot&eacute;olytique de la caspase 8 et 3 dans les cellules SH-SY5Y. Ces r&eacute;sultats ont d&eacute;montr&eacute; A20 pourrait efficacement prot&eacute;ger les neurones contre l&#39;apoptose post-isch&eacute;mique et peuvent fonctionner en partie sur la voie de caspase mort du r&eacute;cepteur. Transfert de g&egrave;nes de l&#39;A20 peut &ecirc;tre une approche prometteuse &agrave; la th&eacute;rapie g&eacute;nique pour isch&eacute;mie c&eacute;r&eacute;brale &agrave; l&#39;avenir.

Effet neuroprotecteur de A20 sur post-isch&eacute;mique relatif &agrave; l&#39;apoptose induite par le TNF.

Ischemia cerebrale focale provoca apoptosi nelle cellule neurali durante il periodo di postischemia. TNF &egrave; criticamente coinvolti in tale apoptosi neuronale mediata dalla caspasi percorsi. A20 &egrave; in grado di inibire apoptosi indotta da TNF in molti tipi di cellule. Tuttavia, poco lavoro &egrave; stata effettuata nel sistema nervoso centrale. Nel presente studio, trasferimento del gene della A20 ha portato riduzione del volume di infarto e miglioramento del deficit neurologico nei ratti di ischemia. Risultati di citometria a fluiscono, DNA e TUNEL assay frammentazione tutti indicato che A20 potrebbe inibire l&#39;apoptosi indotta da TNF sia nei neuroni hippocampal primario ratto e cellule SH-SY5Y. Inoltre, abbiamo trovato che A20 mirati il pathway apoptotico TNF inibendo il clivaggio proteolitico della caspase 8 e 3 nelle cellule SH-SY5Y. Questi dati hanno dimostrato A20 efficacemente potrebbe proteggere i neuroni dall&#39;apoptosi postischemic e pu&ograve; funzionare in parte su via caspase recettore di morte. Trasferimento del gene della A20 pu&ograve; essere un approccio promettente per la terapia genica per ischemia cerebrale in futuro.

Effetto neuroprotettivo di A20 sull&#39;apoptosi postischemic indotta da TNF.

We have previously shown that ASK1-interacting protein 1 (AIP1) transduces tumor necrosis factor-induced ASK1-JNK signaling. Because endoplasmic reticulum (ER) stress activates ASK1-JNK signaling cascade, we investigated the role of AIP1 in ER stress-induced signaling. We created AIP1-deficient mice (AIP1-KO) from which mouse embryonic fibroblasts and vascular endothelial cells were isolated. AIP1-KO cells show dramatic reductions in ER stress-induced, but not oxidative stress-induced, ASK1-JNK activation and cell apoptosis. The ER stress-induced IRE1-JNK/XBP-1 axis, but not the PERK-CHOP1 axis, is blunted in AIP1-KO cells. ER stress induced formation of an AIP1-IRE1 complex, and the PH domain of AIP1 is critical for the IRE1 interaction. Furthermore, reconstitution of AIP1-KO cells with AIP1 wild type, not an AIP1 mutant with a deletion of the PH domain (AIP1-DeltaPH), restores ER stress-induced IRE1-JNK/XBP-1 signaling. AIP1-IRE1 association facilitates IRE1 dimerization, a critical step for activation of IRE1 signaling. More importantly, AIP1-KO mice show impaired ER stress-induced IRE1-dependent signaling in vivo. We conclude that AIP1 is essential for transducing the IRE1-mediated ER stress response.

AIP1 is critical in transducing IRE1-mediated endoplasmic reticulum stress response.

Nous avons d&eacute;j&agrave; montr&eacute; que ASK1-interacting protein 1 (AIP1) transduit induite par le facteur de n&eacute;crose tumorale ASK1-JNK signalisation. Parce que le stress du r&eacute;ticulum endoplasmique (ER) active la cascade de signalisation de ASK1-JNK, nous avons &eacute;tudi&eacute; le r&ocirc;le de AIP1 dans ER stress induit par la signalisation. Nous avons cr&eacute;&eacute; des souris d&eacute;ficientes en AIP1 (AIP1-KO) de quelle souris fibroblastes embryonnaires et les cellules endoth&eacute;liales vasculaires ont &eacute;t&eacute; isol&eacute;s. AIP1-KO cellules montrent des r&eacute;ductions spectaculaires dans ER induite par le stress, mais pas oxydative provoqu&eacute;e par le stress, activation ASK1-JNK et l&#39;apoptose des cellules. L&#39;axe de IRE1-JNK/XBP-1 ER induite par le stress, mais pas l&#39;axe PERK-CHOP1, est affaiblie dans les cellules AIP1-KO. ER stress induit la formation d&#39;un complexe AIP1-IRE1 et le domaine de PH de AIP1 est critique pour l&#39;interaction IRE1. En outre, la reconstitution des cellules AIP1-KO avec AIP1 type sauvage, pas un mutant AIP1 avec une suppression du domaine PH (AIP1-DeltaPH), restaure ER stress induit par la signalisation des IRE1-JNK/XBP-1. Association AIP1-IRE1 facilite IRE1 dim&eacute;risation, une &eacute;tape essentielle pour l&#39;activation de la signalisation IRE1. Plus important encore, souris AIP1-KO montrent avec facult&eacute;s affaiblies ER induite par le stress IRE1-d&eacute;pendante de signalisation in vivo. Nous concluons que AIP1 est essentielle pour la transduction de la r&eacute;ponse au stress ER IRE1-m&eacute;diation.

AIP1 est essentiel dans la transduction de la r&eacute;action de stress IRE1 m&eacute;di&eacute;e par le r&eacute;ticulum endoplasmique.

Abbiamo precedentemente dimostrato che ASK1-interacting protein 1 (AIP1) transduces indotta dal fattore di necrosi tumorale ASK1-JNK segnalazione. Perch&eacute; lo stress del reticolo endoplasmatico (ER) attiva la cascata di segnalazione ASK1-JNK, abbiamo studiato il ruolo di AIP1 in ER stress-induced segnalazione. Abbiamo creato il mouse AIP1-carenti (AIP1-KO) dal quale mouse fibroblasti embrionali e cellule endoteliali vascolari sono state isolate. AIP1-KO cellule mostrano una riduzione drammatica in ER stress-indotta, ma non ossidativo stress-induced, ASK1-JNK attivazione e apoptosi delle cellule. L&#39;ER stress-induced IRE1-JNK/XBP-1 asse, ma non l&#39;asse PERK-CHOP1, &egrave; attenuato in cellule AIP1-KO. ER stress induced formazione di un complesso AIP1-IRE1, e il dominio di PH di AIP1 &egrave; critico per l&#39;interazione di IRE1. Inoltre, la ricostituzione delle cellule AIP1-KO con AIP1 di tipo selvatico, non un mutante di AIP1 con la cancellazione del dominio PH (AIP1-DeltaPH), ripristina ER stress-induced IRE1-JNK/XBP-1 segnalazione. Associazione AIP1-IRE1 facilita la dimerizzazione di IRE1, un passaggio fondamentale per l&#39;attivazione di IRE1 segnalazione. Ancora pi&ugrave; importante, AIP1-KO topi mostrano alterata ER stress-induced IRE1-dipendente dalla segnalazione in vivo. Concludiamo che AIP1 &egrave; essenziale per transducing la risposta allo stress mediata da IRE1 ER.

AIP1 &egrave; un fattore critico nel transducing reticolo endoplasmatico IRE1-mediata risposta allo stress.

Previously we have shown that AIP1 (apoptosis signal-regulating kinase [ASK]1-interacting protein 1), a novel member of the Ras-GAP protein family, facilitates dephosphorylation of ASK1 at pSer967 and subsequently 14-3-3 release from ASK1, leading to enhanced ASK1-JNK signaling. However, the phosphatase(s) responsible for ASK1 dephosphorylation at pSer967 has not been identified. In the present study, we identified protein phosphatase (PP)2A as a potential phosphatase in vascular endothelial cells (ECs). Tumor necrosis factor (TNF)-induced dephosphorylation of ASK1 pSer967 in ECs was blocked by PP2A inhibitor okadaic acid. Overexpression of PP2A catalytic subunit induced dephosphorylation of ASK1 pSer967 and activation of c-Jun N-terminal kinase (JNK). In contrast, a catalytic inactive form of PP2A or PP2A small interfering RNA blunted TNF-induced dephosphorylation of ASK1 pSer967 and activation of JNK without effects on NF-kappaB activation. Whereas AIP1, via its C2 domain, binds to ASK1, PP2A binds to the GAP domain of AIP1. Endogenous AIP1-PP2A complex can be detected in the resting state, and TNF induces a complex formation of AIP1-PP2A with ASK1. Furthermore, TNF-induced association of PP2A with ASK1 was diminished in AIP1-knockdown ECs, suggesting a critical role of AIP1 in recruiting PP2A to ASK1. TNF-signaling molecules TRAF2 and RIP1, known to be in complex with AIP1 and activate AIP1 by phosphorylating AIP1 at Ser604, are critical for TNF-induced ASK1 dephosphorylation. Finally, PP2A and AIP1 cooperatively induce activation of ASK1-JNK signaling and EC apoptosis, as demonstrated by both overexpression and small interfering RNA knockdown approaches. Taken together, our data support a critical role of PP2A-AIP1 complex in TNF-induced activation of ASK1-JNK apoptotic signaling.

AIP1 recruits phosphatase PP2A to ASK1 in tumor necrosis factor-induced ASK1-JNK activation.

Nous l&#39;avons montr&eacute; pr&eacute;c&eacute;demment que AIP1 (apoptosis signal r&eacute;glementant kinase [demande] 1-interacting protein 1), nouveau membre de la famille des prot&eacute;ines Ras-GAP, facilite la d&eacute;phosphorylation du ASK1 au pSer967 et par la suite communiqu&eacute; de 14-3-3 de ASK1, conduisant &agrave; la signalisation am&eacute;lior&eacute;e de ASK1-JNK. Toutefois, le responsable de la d&eacute;phosphorylation de ASK1 &agrave; pSer967 de phosphatase(s) n&#39;a pas &eacute;t&eacute; identifi&eacute;. Dans la pr&eacute;sente &eacute;tude, nous avons identifi&eacute; 2 a de la prot&eacute;ine phosphatase (PP) comme une phosphatase potentielle dans les cellules endoth&eacute;liales vasculaires (ECs). Facteur de n&eacute;crose tumorale (TNF)-induite par d&eacute;phosphorylation du ASK1 pSer967 en ECs a &eacute;t&eacute; bloqu&eacute;e par l&#39;acide okada&iuml;que PP2A inhibiteur. La surexpression de la sous-unit&eacute; catalytique PP2A induite par d&eacute;phosphorylation du ASK1 pSer967 et activation de la kinase c-Jun N-terminal (JNK). En revanche, une forme inactive catalytique PP2A ou petits ARN interf&eacute;rents PP2A &eacute;mouss&eacute; induite par le TNF d&eacute;phosphorylation de ASK1 pSer967 et l&#39;activation des JNK sans effet sur l&#39;activation de NF-kappaB. Consid&eacute;rant que par l&#39;interm&eacute;diaire de son domaine C2, AIP1, se lie &agrave; ASK1, PP2A se lie &agrave; la lacune de AIP1. Endog&egrave;ne AIP1-PP2A complexe peut &ecirc;tre d&eacute;tect&eacute; dans l&#39;&eacute;tat de repos, et TNF induit la formation d&#39;une complexe de AIP1-PP2A avec ASK1. En outre, induite par le TNF association de PP2A avec ASK1 a &eacute;t&eacute; diminu&eacute;e chez ECs AIP1-knockdown, sugg&eacute;rant un r&ocirc;le critique de AIP1 dans recrutement PP2A &agrave; ASK1. Mol&eacute;cules de signalisation TNF TRAF2 et RIP1, connus pour &ecirc;tre en complexe avec AIP1 et activer AIP1 par phosphorylation AIP1 &agrave; Ser604, sont cruciales pour la d&eacute;phosphorylation de l&#39;ASK1 induite par le TNF. Enfin, PP2A et AIP1 en collaboration induisent l&#39;activation de signalisation ASK1-JNK et EC l&#39;apoptose, comme l&#39;a d&eacute;montr&eacute; par surexpression et petites approches knockdown interf&eacute;rentes de RNA. Pris ensemble, les donn&eacute;es supportent un r&ocirc;le critique du complexe PP2A-AIP1 dans l&#39;activation induite par le TNF de signalisation apoptotique de ASK1-JNK.

AIP1 recrute phosphatase PP2A &agrave; ASK1 dans l&#39;activation des JNK-ASK1 induite par le facteur de n&eacute;crose tumorale.

In precedenza abbiamo dimostrato che AIP1 (apoptosi regolazione del segnale chinasi [ASK] 1-interazione proteina 1), un romanzo membro della famiglia della proteina Ras-GAP, facilita la defosforilazione del ASK1 a pSer967 e, successivamente, 14-3-3 sblocco da ASK1, portando a maggiore ASK1-JNK segnalazione. Tuttavia, non &egrave; stato identificato il responsabile della defosforilazione ASK1 a pSer967 phosphatase(s). Nel presente studio, abbiamo individuato le proteine fosfatasi (PP) 2A come una potenziale fosfatasi in cellule endoteliali vascolari (ECs). Fattore di necrosi tumorale (TNF)-indotta defosforilazione del ASK1 pSer967 in ECs &egrave; stato bloccato da acido okadaico di PP2A inibitore. Sovraespressione della subunit&agrave; catalitica di PP2A indotta da attivazione di c-Jun N-terminal chinasi (JNK) e defosforilazione del ASK1 pSer967. Al contrario, una forma inattiva catalitica di PP2A o piccolo RNA interferente PP2A blunted indotta da TNF defosforilazione del ASK1 pSer967 e attivazione di JNK senza effetti sull&#39;attivazione di NF-kappaB. Considerando che AIP1, attraverso il dominio C2, si lega al ASK1, PP2A si lega al dominio di AIP1 GAP. Complesso AIP1-PP2A endogena pu&ograve; essere rilevata nello stato di riposo, e TNF induce una formazione complessa di AIP1-PP2A con ASK1. Inoltre, associazione TNF-indotta di PP2A con ASK1 era diminuito in ECs AIP1-atterramento, suggerendo un ruolo critico di AIP1 nel reclutamento PP2A a ASK1. Molecole di segnalazione TNF TRAF2 e RIP1, conosciuto per essere nel complesso con AIP1 e AIP1 per attivare fosforilante AIP1 a Ser604, sono critiche per defosforilazione ASK1 indotta da TNF. Infine, PP2A e AIP1 in modo cooperativo indurre attivazione di segnalazione ASK1-JNK e CE apoptosi, come dimostrato da iperespressione sia piccoli approcci atterramento interfering RNA. Presi insieme, i nostri dati supportano un ruolo critico di PP2A-AIP1 complesso in attivazione indotta da TNF di segnalazione apoptotic ASK1-JNK.

AIP1 reclute fosfatasi PP2A a ASK1 in attivazione JNK ASK1 indotta dal fattore di necrosi tumorale.

Thioredoxin-2 (Trx2), a major antioxidant protein in mitochondria, enhances nitric oxide bioavailability and inhibits ASK1-dependent apoptosis in endothelial cells (ECs). However, the in vivo role of Trx2 in angiogenesis has not been defined. Here we used EC-specific transgenesis of Trx2 (Trx2-TG) in mice to determine the in vivo function of Trx2 in arteriogenesis and angiogenesis.

Endothelial-specific expression of mitochondrial thioredoxin promotes ischemia-mediated arteriogenesis and angiogenesis.

Thior&eacute;doxine-2 (Trx2), une prot&eacute;ine antioxydante majeure dans les mitochondries, am&eacute;liore la biodisponibilit&eacute; de l&#39;oxyde nitrique et inhibe la ASK1 d&eacute;pendant de l&#39;apoptose dans les cellules endoth&eacute;liales (CE). Cependant, le r&ocirc;le in vivo de Trx2 dans l&#39;angiogen&egrave;se n&#39;a pas &eacute;t&eacute; d&eacute;fini. Ici nous avons utilis&eacute; la transg&eacute;n&egrave;se EC sp&eacute;cifique de Trx2 (Trx2-TG) chez la souris pour d&eacute;terminer la fonction in vivo de Trx2 en arteriogenesis et l&#39;angiog&eacute;n&egrave;se.

Expression endoth&eacute;liales sp&eacute;cifique de thioredoxin mitochondriale favorise l&#39;angiogen&egrave;se et l&#39;arteriogenesis induite par l&#39;isch&eacute;mie.

Tioredossina-2 (Trx2), una proteina importante antiossidante nei mitocondri, aumenta la biodisponibilit&agrave; di ossido nitrico e inibisce l&#39;apoptosi ASK1-dipendente nelle cellule endoteliali (ECs). Tuttavia, non &egrave; stato definito il ruolo in vivo di Trx2 nell&#39;angiogenesi. Qui abbiamo usato la transgenesi CE specifiche di Trx2 (Trx2-TG) nei topi per determinare la funzione in vivo di Trx2 arteriogenesis e l&#39;angiogenesi.

Endoteliali specifica espressione della tioredossina mitocondriale promuove l&#39;angiogenesi e arteriogenesis ischemia-mediata.

Previously we have shown that tyrosine 718 of ASK1 when phosphorylated is critical for SOCS1 binding and SOCS1-mediated degradation of ASK1. However, the kinase and phosphatase responsible for phosphorylation and dephosphorylation of ASK1 at Tyr-718 are unknown. In this study, we identified JAK2 and SHP2 as a Tyr-718-specific kinase and phosphatase, respectively. Interferon-gamma (IFN-gamma) induced degradation of ASK1 in normal but not in SOCS1-KO endothelial cells (EC). IFN-gamma-induced tyrosine phosphorylation of ASK1 at Tyr-718 was blocked by a JAK2-specific inhibitor. IFN-gamma enhanced the association between JAK2 and ASK1, and the ASK1-JAK2 complex was labile and was stabilized by the proteasomal inhibitor MG132. Furthermore, JAK2, but not JAK1, directly bound to and phosphorylated ASK1 at Tyr-718, leading to an enhanced association of ASK1 with SOCS1 and subsequent ASK1 degradation. Next, we showed that overexpression of the SH2-containing protein-tyrosine phosphatase-2 (SHP2) augmented, whereas a phosphatase-inactive mutant of SHP2 inhibited, TNF-induced ASK1 dephosphorylation. SHP2 associated with ASK1 in response to tumor necrosis factor in EC. An SHP-2 substrate-trapping mutant formed a complex with tyrosine-phosphorylated ASK1, suggesting that ASK1 is a direct SHP2 substrate. Moreover, SHP2 wild type, but not a catalytically inactive mutant, dissociated SOCS1 from ASK1. IFN-gamma-induced ASK1 Tyr(P)-718 was enhanced in mouse EC deficient in SHP2 (SHP2-KO). In contrast, tumor necrosis factor-induced dephosphorylation of ASK1 at Tyr(P)-718 and activation of ASK1-JNK signaling, as well as EC apoptosis, are significantly reduced in SHP2-KO EC. Our data suggest that JAK2-SOCS1 and SHP2 reciprocally regulate ASK1 phosphorylation and stability in response to cytokines.

JAK2 and SHP2 reciprocally regulate tyrosine phosphorylation and stability of proapoptotic protein ASK1.

Pr&eacute;c&eacute;demment, nous avons montr&eacute; que la tyrosine 718 de ASK1 lorsque phosphoryl&eacute; est critique pour la liaison SOCS1 et d&eacute;gradation induite par le SOCS1 de ASK1. Cependant, la kinase et la phosphatase responsable de la phosphorylation et la d&eacute;phosphorylation de ASK1 &agrave; Tyr-718 sont inconnus. Dans cette &eacute;tude, nous avons identifi&eacute; JAK2 et SHP2 comme une kinase de Tyr-718-sp&eacute;cifiques et de la phosphatase, respectivement. Interf&eacute;ron-gamma (IFN-gamma) induits par la d&eacute;gradation des ASK1 dans des conditions normales, mais pas dans les cellules endoth&eacute;liales de SOCS1-KO (EC). La phosphorylation de tyrosine induite par l&#39;IFN-gamma de ASK1 &agrave; Tyr-718 &eacute;tait bloqu&eacute;e par un inhibiteur de JAK2 sp&eacute;cifiques. IFN-gamma am&eacute;lior&eacute; l&#39;association entre JAK2 et ASK1, et le complexe ASK1-JAK2 est labile et a &eacute;t&eacute; stabilis&eacute; par l&#39;inhibiteur du prot&eacute;asome MG132. En outre, JAK2, mais pas JAK1, directement li&eacute; &agrave; et phosphoryl&eacute;e ASK1 &agrave; Tyr-718, conduisant &agrave; une association accrue de ASK1 avec SOCS1 et d&eacute;gradation subs&eacute;quente de ASK1. Ensuite, nous avons montr&eacute; que la surexpression du SH2 contenant prot&eacute;ine-tyrosine phosphatase-2 (SHP2) augment&eacute;e, alors qu&#39;un mutant phosphatase-inactif de SHP2 inhib&eacute;e, induite par le TNF ASK1 d&eacute;phosphorylation. Shp2 associ&eacute; &agrave; ASK1 en r&eacute;ponse au facteur de n&eacute;crose tumorale par EC. Un mutant de substrat-pi&eacute;geage SHP-2 forme un complexe avec la tyrosine phosphoryl&eacute;s ASK1, sugg&eacute;rant que le ASK1 est un substrat de SHP2 direct. En outre, SHP2 sauvage, mais pas un mutant catalytiquement inactif, dissoci&eacute; SOCS1 de ASK1. Induite par l&#39;IFN-gamma ASK1 Tyr (P)-718 a &eacute;t&eacute; am&eacute;lior&eacute; dans les souris d&eacute;ficiente en SHP2 EC (SHP2-KO). En revanche, induite par le facteur de n&eacute;crose tumorale d&eacute;phosphorylation de ASK1 &agrave; Tyr (P)-718 et l&#39;activation des JNK-ASK1 de signalisation, ainsi que EC l&#39;apoptose, sont consid&eacute;rablement r&eacute;duits dans SHP2-KO ce. Nos donn&eacute;es sugg&egrave;rent que les JAK2-SOCS1 et SHP2 r&eacute;gulent mutuellement la phosphorylation de ASK1 et de la stabilit&eacute; en r&eacute;ponse &agrave; des cytokines.

JAK2 et SHP2 r&eacute;gulent r&eacute;ciproquement la phosphorylation de tyrosine et de la stabilit&eacute; de la prot&eacute;ine pro-apoptotique ASK1.

In precedenza abbiamo dimostrato che tirosina 718 di ASK1 quando fosforilato &egrave; critico per l&#39;associazione di SOCS1 e SOCS1-mediato degradazione della ASK1. Tuttavia, la chinasi e fosfatasi responsabile della fosforilazione e defosforilazione di ASK1 a Tyr-718 sono sconosciuti. In questo studio, abbiamo identificato JAK2 e SHP2 come un Tyr-718-specifiche chinasi e fosfatasi, rispettivamente. L&#39;interferone-gamma (IFN-gamma) indotta da degradazione di ASK1 in normale, ma non in cellule endoteliali SOCS1-KO (CE). La fosforilazione della tirosina indotta da IFN-gamma di ASK1 a Tyr-718 &egrave; stata bloccata da un inibitore di JAK2-specifici. IFN-gamma rafforzata l&#39;associazione tra JAK2 e ASK1, e il complesso ASK1-JAK2 &egrave; labile e stato stabilizzato con l&#39;inibitore proteosomale MG132. Inoltre, JAK2, ma non JAK1, direttamente associato a e fosforilato ASK1 a Tyr-718, portando ad un&#39;associazione maggiore di ASK1 con SOCS1 e successiva degradazione ASK1. Successivamente, abbiamo mostrato che iperespressione del contenenti SH2 della proteina tirosina fosfatasi-2 (SHP2) aumentata, considerando che un mutante inattivo fosfatasi di SHP2 inibito, indotta da TNF ASK1 dephosphorylation. SHP2 associato ASK1 in risposta al fattore di necrosi tumorale nella CE. Un mutante del substrato-trapping SHP-2 forma un complesso con tirosina fosforilata ASK1, suggerendo che ASK1 &egrave; un substrato SHP2 diretto. Inoltre, SHP2 wild type, ma non un mutante cataliticamente inattivo, dissociato SOCS1 da ASK1. Indotta da IFN-gamma ASK1 Tyr (P)-718 &egrave; stato migliorato nei mouse carenti in SHP2 CE (SHP2-KO). In contrasto, indotta dal fattore di necrosi tumorale defosforilazione di ASK1 a Tyr (P)-718 e l&#39;attivazione di segnalazione ASK1-JNK, cos&igrave; come CE apoptosi, sono significativamente ridotto SHP2-KO CE. I nostri dati suggeriscono che JAK2-SOCS1 e SHP2 reciprocamente regolano la fosforilazione ASK1 e stabilit&agrave; in risposta a citochine.

JAK2 e SHP2 reciprocamente regolano la fosforilazione della tirosina e stabilit&agrave; della proteina Miselli ASK1.

In metastatic prostate cancer (PCa) cells, imbalance between cell survival and death signals such as constitutive activation of phosphatidylinositol 3-kinase (PI3K)-Akt and inactivation of apoptosis-stimulated kinase (ASK1)-JNK pathways is often detected. Here, we show that DAB2IP protein, often down-regulated in PCa, is a potent growth inhibitor by inducing G(0)/G(1) cell cycle arrest and is proapoptotic in response to stress. Gain of function study showed that DAB2IP can suppress the PI3K-Akt pathway and enhance ASK1 activation leading to cell apoptosis, whereas loss of DAB2IP expression resulted in PI3K-Akt activation and ASK1-JNK inactivation leading to accelerated PCa growth in vivo. Moreover, glandular epithelia from DAB2IP(-/-) animal exhibited hyperplasia and apoptotic defect. Structural functional analyses of DAB2IP protein indicate that both proline-rich (PR) and PERIOD-like (PER) domains, in addition to the critical role of C2 domain in ASK1 activity, are important for modulating PI3K-Akt activity. Thus, DAB2IP is a scaffold protein capable of bridging both survival and death signal molecules, which implies its role in maintaining cell homeostasis.

DAB2IP coordinates both PI3K-Akt and ASK1 pathways for cell survival and apoptosis.

Dans les cellules de cancer de la prostate m&eacute;tastatique (PCa), d&eacute;s&eacute;quilibre entre la survie des cellules et la mort des signaux tels que l&#39;activation constitutive de la phosphatidylinositol 3-kinase (PI3K)-Akt et inactivation de la kinase stimul&eacute;e par apoptose (ASK1)-voies de JNK est souvent d&eacute;tect&eacute;. Ici, nous montrons que DAB2IP prot&eacute;ine, souvent diminu&eacute;e dans APC, est un inhibiteur puissant de la croissance en provoquant l&#39;arr&ecirc;t du cycle cellulaire G(0)/G(1) et qu&#39;il est pro-apoptotique en r&eacute;ponse au stress. Gain d&#39;&eacute;tude de la fonction a montr&eacute; que les DAB2IP peuvent supprimer la voie PI3K-Akt et am&eacute;liorer l&#39;activation ASK1 conduisant &agrave; l&#39;apoptose des cellules, tandis que la perte d&#39;expression DAB2IP entra&icirc;nerait PI3K-Akt activation et l&#39;inactivation de ASK1-JNK conduisant &agrave; la croissance acc&eacute;l&eacute;r&eacute;e des PCa in vivo. En outre, les &eacute;pith&eacute;liums glandulaires de DAB2IP(-/-) animal expos&eacute;es hyperplasie et apoptotiques d&eacute;faut. Structurales analyses fonctionnelles des prot&eacute;ines DAB2IP indiquent que la proline-riche (PR) et domaines comme p&eacute;riode (PER), outre le r&ocirc;le essentiel du domaine C2 ASK1 activit&eacute;, sont importants pour la modulation de l&#39;activit&eacute; PI3K-Akt. Ainsi, DAB2IP est une prot&eacute;ine d&#39;&eacute;chafaudage capable de combler la survie et mol&eacute;cules de signal de mort, ce qui implique son r&ocirc;le dans le maintien de l&#39;hom&eacute;ostasie cellulaire.

DAB2IP coordonne les PI3K-Akt et ASK1 les voies de la survie cellulaire et l&#39;apoptose.

Nelle cellule di cancro metastatico della prostata (PCa), i segnali di squilibrio tra la sopravvivenza delle cellule e la morte come attivazione costitutiva del fosfatidilinositolo 3-chinasi (PI3K)-Akt e inattivazione di apoptosi stimolata chinasi (ASK1)-percorsi di JNK &egrave; spesso rilevato. Mostriamo qui, che DAB2IP proteine, spesso gi&ugrave;-regolamentata in PCa, sono un inibitore potente crescita inducendo G(0)/G(1) arresto del ciclo cellulare ed sono Miselli in risposta allo stress. Guadagno di studio di funzione ha mostrato che DAB2IP pu&ograve; sopprimere il pathway PI3K-Akt e migliorare ASK1 activation che porta all&#39;apoptosi delle cellule, considerando che la perdita di espressione di DAB2IP ha provocato PI3K-Akt attivazione ed inattivazione ASK1-JNK che porta alla crescita accelerata PCa in vivo. Inoltre, epiteli ghiandolari da DAB2IP(-/-) animale esposto iperplasia e apoptotic difetto. Analisi strutturali funzionali della proteina DAB2IP indicano che sia ricco di prolina (PR) e domini simili a periodo (PER), oltre al ruolo critico del dominio C2 ASK1 attivit&agrave;, sono importanti per la modulazione attivit&agrave; PI3K-Akt. Cos&igrave;, DAB2IP &egrave; una proteina dell&#39;impalcatura in grado di colmare la sopravvivenza sia molecole segnale di morte, che implica il suo ruolo nel mantenimento dell&#39;omeostasi cellulare.

DAB2IP coordinate sia PI3K-Akt e ASK1 percorsi per la sopravvivenza delle cellule e apoptosis.

A single nucleotide polymorphism in the DAB2IP gene is associated with risk of aggressive prostate cancer (PCa), and loss of DAB2IP expression is frequently detected in metastatic PCa. However, the functional role of DAB2IP in PCa remains unknown. Here, we show that the loss of DAB2IP expression initiates epithelial-to-mesenchymal transition (EMT), which is visualized by repression of E-cadherin and up-regulation of vimentin in both human normal prostate epithelial and prostate carcinoma cells as well as in clinical prostate-cancer specimens. Conversely, restoring DAB2IP in metastatic PCa cells reversed EMT. In DAB2IP knockout mice, prostate epithelial cells exhibited elevated mesenchymal markers, which is characteristic of EMT. Using a human prostate xenograft-mouse model, we observed that knocking down endogenous DAB2IP in human carcinoma cells led to the development of multiple lymph node and distant organ metastases. Moreover, we showed that DAB2IP functions as a scaffold protein in regulating EMT by modulating nuclear beta-catenin/T-cell factor activity. These results show the mechanism of DAB2IP in EMT and suggest that assessment of DAB2IP may provide a prognostic biomarker and potential therapeutic target for PCa metastasis.

Role of DAB2IP in modulating epithelial-to-mesenchymal transition and prostate cancer metastasis.

Un polymorphisme simple nucl&eacute;otide dans le g&egrave;ne DAB2IP est associ&eacute; avec le risque de cancer de la prostate agressif (PCa), et la perte d&#39;expression DAB2IP est fr&eacute;quemment d&eacute;tect&eacute; dans APC m&eacute;tastatique. Cependant, le r&ocirc;le fonctionnel de DAB2IP dans APC demeure inconnu. Ici, nous montrons que la perte d&#39;expression DAB2IP lance transition &eacute;pith&eacute;liale-m&eacute;senchymateuse (EMT), qui est visualis&eacute;e par la r&eacute;pression de la E-cadh&eacute;rine et r&eacute;gulation de vimentine dans les deux cellules humaine normale &eacute;pith&eacute;lial et la prostate cancer de la prostate, ainsi que dans les &eacute;chantillons cliniques de cancer de la prostate. &Agrave; l&#39;inverse, restauration DAB2IP dans les cellules m&eacute;tastatiques de PCa invers&eacute; EMT. Chez les souris knock-out DAB2IP, cellules &eacute;pith&eacute;liales prostatiques expos&eacute; marqueurs m&eacute;senchymateuses &eacute;lev&eacute;es, qui est caract&eacute;ristique de l&#39;EMT. &Agrave; l&#39;aide d&#39;un mod&egrave;le de x&eacute;nogreffe de la prostate humain-souris, nous avons observ&eacute; que faire tomber DAB2IP endog&egrave;ne dans les cellules de carcinome humain conduit &agrave; l&#39;&eacute;laboration de plusieurs ganglions lymphatiques et m&eacute;tastases organes &eacute;loign&eacute;s. En outre, nous avons montr&eacute; que DAB2IP fonctionne comme une prot&eacute;ine d&#39;&eacute;chafaudage dans la r&eacute;gulation des EMT en modulant l&#39;activit&eacute; de facteur b&ecirc;ta-cat&eacute;nine/T-cellule nucl&eacute;aire. Ces r&eacute;sultats montrent le m&eacute;canisme de DAB2IP en EMT et sugg&egrave;rent que l&#39;&eacute;valuation de DAB2IP peut fournir un biomarqueur pronostique et une cible th&eacute;rapeutique potentielle pour APC m&eacute;tastases.

R&ocirc;le de DAB2IP dans la modulation de transition &eacute;pith&eacute;liale-m&eacute;senchymateuse et m&eacute;tastase de cancer de la prostate.

Un polimorfismo a singolo nucleotide nel gene DAB2IP &egrave; associato con il rischio di aggressivo cancro della prostata (PCa) e perdita di espressione di DAB2IP &egrave; spesso rilevato in PCa metastatico. Tuttavia, il ruolo funzionale di DAB2IP in PCa rimane sconosciuto. Qui, ci mostra che la perdita di espressione DAB2IP avvia la transizione epitelio mesenchimale (EMT), che viene visualizzato dalla repressione della E-caderina e up-regolazione di vimentina nelle cellule sia umano normale della prostata epiteliale e della prostata carcinoma cos&igrave; come in campioni clinici di cancro alla prostata. Al contrario, ripristino di DAB2IP nelle cellule APC metastatiche invertito EMT. DAB2IP topi knockout, cellule epiteliali prostatiche esposta elevata marker mesenchimale, che &egrave; caratteristica della EMT. Utilizzando un modello umano della prostata del mouse xenoinnesto, abbiamo osservato che abbattendo DAB2IP endogena in cellule di carcinoma umano ha portato allo sviluppo di multipli linfonodi e metastasi in organi distanti. Inoltre, abbiamo dimostrato che DAB2IP funziona come una proteina dell&#39;impalcatura nella regolazione della EMT modulando l&#39;attivit&agrave; del fattore nucleare di beta-catenina/T-cell. Questi risultati mostrano il meccanismo di DAB2IP in EMT e suggeriscono che valutazione del DAB2IP pu&ograve; fornire un biomarcatore prognostico e un potenziale bersaglio terapeutico per PCa metastasi.

Ruolo di DAB2IP nel modulare la transizione epitelio mesenchimale e metastasi del cancro alla prostata.

Cerebral cavernous malformations (CCMs) are human vascular malformations caused by mutations in three genes of unknown function: CCM1, CCM2, and CCM3. CCM3, also known as PDCD10 (programmed cell death 10), was initially identified as a messenger RNA whose abundance was induced by apoptotic stimuli in vitro. However, the in vivo function of CCM3 has not been determined. Here, we describe mice with a deletion of the CCM3 gene either ubiquitously or specifically in the vascular endothelium, smooth muscle cells, or neurons. Mice with global or endothelial cell-specific deletion of CCM3 exhibited defects in embryonic angiogenesis and died at an early embryonic stage. CCM3 deletion reduced vascular endothelial growth factor receptor 2 (VEGFR2) signaling in embryos and endothelial cells. In response to VEGF stimulation, CCM3 was recruited to and stabilized VEGFR2, and the carboxyl-terminal domain of CCM3 was required for the stabilization of VEGFR2. Indeed, the CCM3 mutants found in human patients lacking the carboxyl-terminal domain were labile and were unable to stabilize and activate VEGFR2. These results demonstrate that CCM3 promotes VEGFR2 signaling during vascular development.

Stabilization of VEGFR2 signaling by cerebral cavernous malformation 3 is critical for vascular development.

Cerebral cavernous malformations (CCMs) sont des malformations vasculaires humaines caus&eacute;es par des mutations dans trois g&egrave;nes de fonction inconnue : CCM1, CCM2 et CCM3. CCM3, &eacute;galement connu sous le nom de PDCD10 (mort cellulaire programm&eacute;e, 10), a &eacute;t&eacute; initialement identifi&eacute; comme un ARN messager dont l&#39;abondance est induite par des stimuli apoptotiques in vitro. Cependant, la fonction in vivo de CCM3 n&#39;a pas &eacute;t&eacute; d&eacute;termin&eacute;e. Nous d&eacute;crivons ici, souris avec une d&eacute;l&eacute;tion du g&egrave;ne CCM3 partout ou plus pr&eacute;cis&eacute;ment dans l&#39;endoth&eacute;lium vasculaire, les cellules musculaires lisses ou des neurones. Souris avec globale ou endoth&eacute;liale suppression sp&eacute;cifique des cellules de CCM3 pr&eacute;sentaient des d&eacute;fauts dans l&#39;angiogen&egrave;se embryonnaires et mourut &agrave; un stade embryonnaire pr&eacute;coce. CCM3 suppression r&eacute;duit vascular endothelial growth factor receptor 2 (VEGFR2) la signalisation dans les embryons et les cellules endoth&eacute;liales. En r&eacute;ponse &agrave; la stimulation de VEGF, CCM3 a &eacute;t&eacute; recrut&eacute; &agrave; et stabilis&eacute; VEGFR2 et le domaine c-terminal de CCM3 &eacute;tait n&eacute;cessaire pour la stabilisation de VEGFR2. En effet, les mutants CCM3 trouv&eacute;s chez des patients humains manquent le domaine c-terminal sont labiles et ne purent se stabilise et VEGFR2. Ces r&eacute;sultats d&eacute;montrent que le CCM3 favorise VEGFR2 signalisation au cours du d&eacute;veloppement vasculaire.

Stabilisation de la VEGFR2 signalisation par malformation caverneuse c&eacute;r&eacute;brale 3 est critique pour le d&eacute;veloppement vasculaire.

Malformazioni cavernose cerebrali (CCM) sono malformazioni vascolari umane causate da mutazioni di tre geni della funzione sconosciuta: CCM1, CCM2 e CCM3. CCM3, noto anche come PDCD10 (morte cellulare programmata, 10), &egrave; stato inizialmente identificato come un RNA messaggero cui abbondanza &egrave; stata indotta da stimoli apoptotic in vitro. Tuttavia, non &egrave; stata determinata la funzione in vivo di CCM3. Qui, descriviamo i topi con una delezione del gene CCM3 ubiquitously o specificamente l&#39;endotelio vascolare, cellule muscolari lisce o neuroni. Topi con eliminazione di specifici delle cellule endoteliali o globale di CCM3 esposto difetti di angiogenesi embrionale e morirono in una fase embrionale precoce. Eliminazione CCM3 ridotto vascular endothelial growth factor receptor 2 (VEGFR2) segnalazione in embrioni e cellule endoteliali. In risposta alla stimolazione di VEGF, CCM3 &egrave; stato reclutato e stabilizzato VEGFR2, e il dominio carbossi-terminale di CCM3 &egrave; stato richiesto per la stabilizzazione del VEGFR2. Infatti, i mutanti CCM3 trovati in pazienti umani manca il dominio carbossi-terminale erano labili e non erano in grado di stabilizzare e attivare VEGFR2. Questi risultati dimostrano che CCM3 promuove VEGFR2 segnalazione durante lo sviluppo vascolare.

Stabilizzazione del VEGFR2 segnalazione da Malformazione cavernosa cerebrale 3 &egrave; fondamentale per lo sviluppo vascolare.

&#x86CB;&#x767D; (SUMOylation) &#x548C; deSUMOylation &#x7684;&#x5C0F;&#x6CDB;&#x7D20;&#x6837;&#x4FEE;&#x9970;&#x7B26; &#xFF08;SUMO&#xFF09; &#x4FEE;&#x6539;&#x5DF2;&#x6210;&#x4E3A;&#x86CB;&#x767D;&#x8D28;&#x529F;&#x80FD;&#x7684;&#x91CD;&#x8981;&#x76D1;&#x7BA1;&#x673A;&#x5236;&#x3002;SENP1 (&#x76F8;&#x6251;&#x7279;&#x5B9A;&#x86CB;&#x767D;&#x9176;) deconjugates SUMOs &#x4ECE;&#x4FEE;&#x6539;&#x540E;&#x7684;&#x86CB;&#x767D;&#x8D28;&#x3002;&#x6211;&#x4EEC;&#x521B;&#x5EFA;&#x4E86;&#x57FA;&#x4E8E; Cre-loxp &#x5E8F;&#x5217;&#x91CD;&#x7EC4;&#x7CFB;&#x7EDF;&#x7684; SENP1 (KO) &#x6572;&#x9664;&#x5C0F;&#x9F20;&#x3002;&#x5168;&#x7403;&#x5220;&#x9664;&#x7684; SENP1 (SENP1 KO&#xFF09; &#x5BFC;&#x81F4;&#x8D2B;&#x8840;&#x548C;&#x80DA;&#x80CE;&#x81F4;&#x6B7B;&#x7387; 13.5 &#x80DA;&#x80CE;&#x5929;&#x548C;&#x4EA7;&#x540E;&#x7B2C;&#x4E00;&#x5929;&#x4E4B;&#x95F4;&#x5173;&#x8054;&#x7EA2;&#x7EC6;&#x80DE;&#x751F;&#x6210;&#x7F3A;&#x9677;&#x80CE;&#x513F;&#x809D;&#x810F;&#x4E2D;&#x3002;&#x9AA8;&#x9AD3;&#x79FB;&#x690D; SENP1 KO &#x80CE;&#x809D;&#x7EC6;&#x80DE;&#x5BF9;&#x8F90;&#x7167;&#x6210;&#x4EBA;&#x6536;&#x4EF6;&#x4EBA;&#x6388;&#x4E88;&#x7EA2;&#x7EC6;&#x80DE;&#x751F;&#x6210;&#x7F3A;&#x9677;&#x3002;&#x86CB;&#x767D;&#x5206;&#x6790;&#x663E;&#x793A; GATA1 &#x548C; GATA1 &#x76F8;&#x5173;&#x57FA;&#x56E0;&#x4E0B;&#x8C03; SENP1 KO &#x5C0F;&#x9F20;&#x80CE;&#x513F;&#x809D;&#x810F;&#x4E2D;&#x3002;&#x8FD9;&#x4E0B;&#x8C03;&#x63A7;&#x4E0E;&#x79EF;&#x7D2F;&#x7684; GATA1 SUMOylated &#x7A97;&#x4F53;&#x7684;&#x5173;&#x8054;&#x3002;&#x6211;&#x4EEC;&#x8FDB;&#x4E00;&#x6B65;&#x8BC1;&#x660E; SENP1 &#x53EF;&#x4EE5;&#x76F4;&#x63A5; deSUMOylate GATA1&#xFF0C;GATA1 &#x76F8;&#x5173;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x548C;&#x7EA2;&#x7EC6;&#x80DE;&#x751F;&#x6210;&#x7684;&#x4F53;&#x5916;&#x6D4B;&#x5B9A;&#x89C4;&#x7BA1;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6211;&#x4EEC;&#x53EF;&#x4EE5;&#x5C55;&#x793A; GATA1 SUMOylation &#x6539;&#x53D8;&#x53CA;&#x5176; DNA &#x7ED3;&#x5408;&#xFF0C;&#x51CF;&#x5C11;&#x5176;&#x62DB;&#x52DF; GATA1 &#x53CD;&#x5E94;&#x57FA;&#x56E0;&#x542F;&#x52A8;&#x5B50;&#x3002;&#x96C6;&#x4F53;&#xFF0C;&#x6211;&#x4EEC;&#x5F97;&#x51FA;&#x7684;&#x7ED3;&#x8BBA; SENP1 &#x7684; deSUMOylating GATA1 &#x4FC3;&#x8FDB; GATA1 &#x6FC0;&#x6D3B;&#x548C;&#x968F;&#x540E;&#x7684;&#x7EA2;&#x7EC6;&#x80DE;&#x751F;&#x6210;&#x3002;

&#x4ECB;&#x5BFC; SENP1 GATA1 deSUMOylation &#x662F;&#x7EA2;&#x7EC6;&#x80DE;&#x751F;&#x6210;&#x6700;&#x7EC8;&#x7684;&#x5173;&#x952E;&#x3002;

Small ubiquitin-like modifier (SUMO) modification of proteins (SUMOylation) and deSUMOylation have emerged as important regulatory mechanisms for protein function. SENP1 (SUMO-specific protease) deconjugates SUMOs from modified proteins. We have created SENP1 knockout (KO) mice based on a Cre-loxP system. Global deletion of SENP1 (SENP1 KO) causes anemia and embryonic lethality between embryonic day 13.5 and postnatal day 1, correlating with erythropoiesis defects in the fetal liver. Bone marrow transplantation of SENP1 KO fetal liver cells to irradiated adult recipients confers erythropoiesis defects. Protein analyses show that the GATA1 and GATA1-dependent genes are down-regulated in fetal liver of SENP1 KO mice. This down-regulation correlates with accumulation of a SUMOylated form of GATA1. We further show that SENP1 can directly deSUMOylate GATA1, regulating GATA1-dependent gene expression and erythropoiesis by in vitro assays. Moreover, we demonstrate that GATA1 SUMOylation alters its DNA binding, reducing its recruitment to the GATA1-responsive gene promoter. Collectively, we conclude that SENP1 promotes GATA1 activation and subsequent erythropoiesis by deSUMOylating GATA1.

SENP1-mediated GATA1 deSUMOylation is critical for definitive erythropoiesis.

Modification de petite ubiquitin-comme modificateur (SUMO) de prot&eacute;ines (SUMOylation) et deSUMOylation sont apparus comme des m&eacute;canismes de r&eacute;glementation importants pour la fonction de prot&eacute;ine. SENP1 (prot&eacute;ase sp&eacute;cifique SUMO) d&eacute;conjugue SUMOs de prot&eacute;ines modifi&eacute;es. Nous avons cr&eacute;&eacute; des souris knockout (KO) de SENP1 bas&eacute;s sur un syst&egrave;me de Cre-loxP. Suppression globale de SENP1 (SENP1 KO) provoque une an&eacute;mie et une l&eacute;talit&eacute; embryonnaire entre embryonnaires 13,5 et postnatal jour 1, corr&eacute;lation avec les d&eacute;fauts de l&#39;&eacute;rythropo&iuml;&egrave;se dans le foie f&oelig;tal. Greffe de moelle osseuse des cellules du foie foetales SENP1 KO aux b&eacute;n&eacute;ficiaires adultes irradi&eacute;s conf&egrave;re les d&eacute;fauts de l&#39;&eacute;rythropo&iuml;&egrave;se. Les analyses de prot&eacute;ines montrent que les g&egrave;nes GATA1 et GATA1 d&eacute;pendantes sont diminu&eacute;es dans le foie des f&oelig;tus de souris SENP1 KO. Cette r&eacute;gulation est en corr&eacute;lation avec l&#39;accumulation d&#39;une forme de SUMOylated de GATA1. De plus, nous montrons que SENP1 peut directement deSUMOylate GATA1, r&eacute;glementant l&#39;expression du g&egrave;ne GATA1 d&eacute;pendante et &eacute;rythropo&iuml;&egrave;se par des essais in vitro. De plus, nous d&eacute;montrons que GATA1 SUMOylation modifie sa liaison &agrave; l&#39;ADN, r&eacute;duisant son recrutement pour le promoteur du g&egrave;ne GATA1-sensible. Collectivement, nous concluons que le SENP1 favorise GATA1 activation et &eacute;rythropo&iuml;&egrave;se ult&eacute;rieur par deSUMOylating GATA1.

SENP1-mediated GATA1 deSUMOylation est critique pour l&#39;&eacute;rythropo&iuml;&egrave;se d&eacute;finitif.

Kleine Ubiquitin-&auml;hnliche-Modifizierer (SUMO) &Auml;nderung der Proteine (SUMOylation) und DeSUMOylation sind entstanden, als wichtige regulatorische Mechanismen f&uuml;r Proteinfunktion. SENP1 (SUMO-spezifische Protease) deconjugates SUMOs aus modifizierte Proteine. Wir haben SENP1 Knockout (KO) M&auml;use basiert auf einem Cre-LoxP-System geschaffen. Globale L&ouml;schung des SENP1 (SENP1 KO) verursacht An&auml;mie und embryonale T&ouml;dlichkeit zwischen embryonalen Tag 13,5 und postnatale Tag 1, Korrelation mit Erythropoese-M&auml;ngel in der fetalen Leber. Knochenmark-Transplantation von SENP1 KO fetalen Leberzellen an bestrahlten Erwachsenen Empf&auml;nger &uuml;bertr&auml;gt Erythropoese M&auml;ngel. Protein-Analysen zeigen, dass die Gene GATA1 und GATA1-abh&auml;ngige unten-geregelt in der fetalen Leber von M&auml;usen SENP1 KO. Diese Herunterregulation korreliert mit Akkumulation einer SUMOylated Form der GATA1. Weiter zeigen wir, dass SENP1 k&ouml;nnen direkt DeSUMOylate GATA1, Regulierung GATA1-abh&auml;ngigen Genexpression und Erythropoese durch in-vitro-Tests. Dar&uuml;ber hinaus zeigen wir, dass GATA1 SUMOylation seine DNA-Bindung &auml;ndert Verringerung ihrer Rekrutierung f&uuml;r den GATA1-responsive gen-F&ouml;rderer. Kollektiv, schlie&szlig;en wir, dass SENP1 DeSUMOylating GATA1 GATA1-Aktivierung und anschlie&szlig;ende Erythropoese f&ouml;rdert.

SENP1-vermittelte GATA1 DeSUMOylation ist entscheidend f&uuml;r die endg&uuml;ltige Erythropoese.

Modifica Small ubiquitin-like modifier (SUMO) di proteine (SUMOylation) e deSUMOylation sono emersi come importanti meccanismi di regolamentazione per la funzione della proteina. SENP1 (proteasi specifiche SUMO) deconjugates Sumo da proteine modificate. Abbiamo creato SENP1 topi knockout (KO) basati su un sistema Cre-loxP. Eliminazione globale di SENP1 (SENP1 KO) provoca l&#39;anemia e la letalit&agrave; embrionale tra giorno embrionale 13,5 e postnatale giorno 1, correlazione con difetti l&#39;eritropoiesi nel fegato fetale. Trapianto di midollo osseo di KO SENP1 cellule epatiche fetali ai destinatari adulti irradiati conferisce l&#39;eritropoiesi difetti. Proteina analisi mostrano che i geni GATA1 e GATA1-dipendente sono gi&ugrave;-regolato nel fegato fetale di topi KO SENP1. Questa down-regulation correla con accumulo di una forma SUMOylated di GATA1. Ulteriormente, ci mostra che SENP1 pu&ograve; direttamente deSUMOylate GATA1, che regolano l&#39;espressione di gene GATA1-dipendente e l&#39;eritropoiesi da saggi in vitro. Inoltre, dimostriamo che GATA1 SUMOylation altera il DNA binding, riducendo la sua assunzione al promotore gene GATA1-sensibli a reagire. Collettivamente, concludiamo che SENP1 promuove GATA1 attivazione e conseguente eritropoiesi da deSUMOylating GATA1.

SENP1-mediata GATA1 deSUMOylation &egrave; fondamentale per l&#39;eritropoiesi definitivo.

&#x5C0F;&#x3055;&#x306A;&#x30E6;&#x30D3;&#x30AD;&#x30C1;&#x30F3;&#x69D8;&#x4FEE;&#x98FE; (&#x76F8;&#x64B2;) &#x4FEE;&#x98FE;&#x86CB;&#x767D;&#x8CEA; (sumo &#x5316;) &#x304A;&#x3088;&#x3073; deSUMOylation &#x306E;&#x91CD;&#x8981;&#x306A;&#x8ABF;&#x7BC0;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x86CB;&#x767D;&#x8CEA;&#x6A5F;&#x80FD;&#x3068;&#x3057;&#x3066;&#x6D6E;&#x4E0A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;SENP1 &#xFF08;&#x76F8;&#x64B2;&#x56FA;&#x6709;&#x30D7;&#x30ED;&#x30C6;&#x30A2;&#x30FC;&#x30BC;&#xFF09; deconjugates &#xFF08;%31anhway&#xFF09; &#x304B;&#x3089;&#x306E;&#x5909;&#x66F4;&#x306E;&#x86CB;&#x767D;&#x8CEA;&#x3002;&#x6211;&#x3005; &#x306F;&#x3001;Cre-loxp &#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306B;&#x57FA;&#x3065;&#x304F; SENP1 &#x30CE;&#x30C3;&#x30AF;&#x30A2;&#x30A6;&#x30C8; (KO) &#x30DE;&#x30A6;&#x30B9;&#x3092;&#x4F5C;&#x6210;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;SENP1 &#x306E;&#x30B0;&#x30ED;&#x30FC;&#x30D0;&#x30EB;&#x524A;&#x9664; (SENP1 KO&#xFF09; &#x539F;&#x56E0;&#x8CA7;&#x8840;&#x304A;&#x3088;&#x3073;&#x840C;&#x82BD;&#x671F;&#x65E5; 13.5 &#x3068;&#x751F;&#x5F8C; 1 &#x5E74;&#x9593;&#x306E;&#x80DA;&#x81F4;&#x6B7B;&#x8D64;&#x8840;&#x7403;&#x6B20;&#x9665;&#x306F;&#x80CE;&#x5150;&#x306E;&#x809D;&#x81D3;&#x306B;&#x76F8;&#x95A2;&#x95A2;&#x9023;&#x3065;&#x3051;&#x308B;&#x3053;&#x3068;&#x3002;&#x9AA8;&#x9AC4;&#x79FB;&#x690D; SENP1 KO &#x80CE;&#x5150;&#x809D;&#x7D30;&#x80DE;&#x7167;&#x5C04;&#x30A2;&#x30C0;&#x30EB;&#x30C8;&#x53D7;&#x4FE1;&#x8005;&#x3078;&#x306E;&#x8D64;&#x8840;&#x7403;&#x306E;&#x6B20;&#x9665;&#x3092;&#x4ED8;&#x4E0E;&#x3057;&#x307E;&#x3059;&#x3002;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x89E3;&#x6790; GATA1 &#x3068; GATA1 &#x4F9D;&#x5B58;&#x578B;&#x907A;&#x4F1D;&#x5B50; SENP1 KO &#x30DE;&#x30A6;&#x30B9;&#x306E;&#x80CE;&#x5150;&#x306E;&#x809D;&#x81D3;&#x3067;&#x30C0;&#x30A6;&#x30F3;&#x898F;&#x5236;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;. &#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x30C0;&#x30A6;&#x30F3;&#x898F;&#x5236; GATA1 &#x306E; SUMOylated &#x30D5;&#x30A9;&#x30FC;&#x30E0;&#x306E;&#x84C4;&#x7A4D;&#x3068;&#x95A2;&#x9023;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B; &#x300E; senp1 &#x300F; &#x3068;&#x76F4;&#x63A5;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059; deSUMOylate GATA1&#x3001;GATA1 &#x4F9D;&#x5B58;&#x578B;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x3068;&#x8D64;&#x8840;&#x7403;&#x306B;&#x3088;&#x308B; in vitro &#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x3092;&#x898F;&#x5236;&#x3059;&#x308B;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x6211;&#x3005; &#x305D;&#x306E; GATA1 &#x5FDC;&#x7B54;&#x6027;&#x907A;&#x4F1D;&#x5B50;&#x30D7;&#x30ED;&#x30E2;&#x30FC;&#x30BF;&#x30FC;&#x306E;&#x52DF;&#x96C6;&#x3092;&#x6E1B;&#x3089;&#x3059; GATA1 sumo &#x5316;&#x304C;&#x305D;&#x306E; DNA &#x306E;&#x7D50;&#x5408;&#x3092;&#x5909;&#x66F4;&#x3055;&#x308C;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x4EE5;&#x4E0B;&#x3001;&#x7DCF;&#x79F0;&#x3057;&#x3066;&#x6211;&#x3005; &#x306F;&#x7D50;&#x8AD6; SENP1 deSUMOylating GATA1 GATA1 &#x6D3B;&#x6027;&#x5316;&#x3068;&#x5F8C;&#x7D9A;&#x306E;&#x9020;&#x8840;&#x3092;&#x4FC3;&#x9032;&#x3057;&#x307E;&#x3059;&#x3002;

SENP1 &#x3092;&#x4ECB;&#x3057;&#x305F; GATA1 deSUMOylation &#x7121;&#x6838;&#x8D64;&#x8840;&#x7403;&#x9020;&#x8840;&#x306B;&#x91CD;&#x8981;&#x3067;&#x3059;&#x3002;

&#xC791;&#xC740; &#xC720;&#xBE44;&#xD034;&#xD2F4; &#xAC19;&#xC740; &#xD55C;&#xC815;&#xC790; (&#xC2A4;&#xBAA8;) &#xC218;&#xC815; (SUMOylation) &#xB2E8;&#xBC31;&#xC9C8;&#xACFC; Desumoylation&#xC758; &#xB2E8;&#xBC31;&#xC9C8; &#xAE30;&#xB2A5;&#xC5D0; &#xB300; &#xD55C; &#xC911;&#xC694; &#xD55C; &#xADDC;&#xC81C; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC73C;&#xB85C; &#xB5A0;&#xC624;&#xB974;&#xACE0; &#xC788;&#xB2E4;. SENP1 (&#xC2A4;&#xBAA8; &#xAD00;&#xB828; protease) &#xC218;&#xC815;&#xB41C; &#xB41C; &#xB2E8;&#xBC31;&#xC9C8;&#xC5D0;&#xC11C; SUMOs deconjugates. &#xC6B0;&#xB9AC;&#xB294; SENP1 &#xB179;&#xC544;&#xC6C3; (KO) &#xB9C8;&#xC6B0;&#xC2A4; Cre loxP &#xC2DC;&#xC2A4;&#xD15C;&#xC744; &#xAE30;&#xBC18;&#xC73C;&#xB85C; &#xB9CC;&#xB4E4;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. Senp1&#xC758; &#xAE00;&#xB85C;&#xBC8C; &#xC0AD;&#xC81C; &#xD0DC;&#xC544; &#xAC04;&#xC5D0;&#xC11C; &#xC801;&#xD608;&#xAD6C; &#xACB0;&#xD568;&#xACFC; &#xBE48; &#xD608;&#xACFC; &#xBC30;&#xC544; &#xCE58; &#xC0AC; &#xC728; 13.5 &#xBC30;&#xC544; &#xC77C;&#xACFC; &#xC0B0; &#xD6C4; &#xD558;&#xB8E8;&#xC5D0; 1 &#xC77C; &#xC0AC;&#xC774; &#xBC1C;&#xC0DD; (SENP1 &#xCF54;). &#xBC29;&#xC0AC;&#xB2A5;&#xB41C; &#xC131;&#xC778; &#xBC1B;&#xB294; &#xC0AC;&#xB78C;&#xC5D0; &#xAC8C; &#xD0DC;&#xC544; &#xAC04; &#xC140; SENP1 &#xCF54;&#xC758; &#xACE8; &#xC218; &#xC774;&#xC2DD; &#xC801;&#xD608;&#xAD6C; &#xACB0;&#xD568;&#xC744; &#xC218; &#xC5EC; &#xD569;&#xB2C8;&#xB2E4;. &#xB2E8;&#xBC31;&#xC9C8; &#xBD84;&#xC11D; &#xB2E4;&#xC6B4; SENP1 &#xCF54; &#xC950;&#xC758; &#xD0DC;&#xC544; &#xAC04;&#xC5D0; &#xB808; &#xADE4; &#xB808;&#xC774;&#xD2B8; &#xB41C; GATA1 &#xBC0F; GATA1 &#xC885;&#xC18D; &#xC720;&#xC804;&#xC790;&#xB294; &#xBCF4;&#xC5EC;&#xC900;&#xB2E4;. &#xC774; &#xB2E4;&#xC6B4; &#xB808; &#xADE4; &#xB808;&#xC774; &#xC158; SUMOylated &#xD615;&#xD0DC;&#xC758; gata1&#xC758; &#xCD95;&#xC801;&#xACFC; &#xC5F0;&#xAD00; &#xC2DC;&#xD0B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; SENP1 &#xC218; &#xC9C1;&#xC811; deSUMOylate GATA1, GATA1 &#xC885;&#xC18D; &#xC720;&#xC804;&#xC790; &#xBC1C;&#xD604; &#xBC0F; &#xC801;&#xD608;&#xAD6C; &#xC0DD;&#xCCB4; &#xC678; &#xBD84;&#xC11D; &#xC2E4;&#xD5D8;&#xC5D0; &#xC758;&#xD574; &#xADDC;&#xC81C; &#xB354; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, GATA1 SUMOylation &#xC790;&#xC0AC;&#xC758; DNA &#xBC14;&#xC778;&#xB529; GATA1 &#xBC18;&#xC751; &#xC720;&#xC804;&#xC790; &#xD504;&#xB85C;&#xBAA8;&#xD130;&#xC758; &#xCC44;&#xC6A9; &#xAC10;&#xC18C; &#xBCC0;&#xACBD; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. &#xC0E8; &#xB2E4;, &#xC6B0;&#xB9AC;&#xB294; SENP1 GATA1 &#xD65C;&#xC131;&#xD654; &#xBC0F; &#xD6C4;&#xC18D; &#xC801;&#xD608;&#xAD6C; deSUMOylating gata1&#xC5D0; &#xC758;&#xD574; &#xCD09;&#xC9C4; &#xACB0;&#xB860; &#xC9C0;&#xC5C8;&#xB2E4;.

GATA1 deSUMOylation SENP1 &#xC911;&#xC7AC; &#xD655;&#xC2E4; &#xD55C; &#xC801;&#xD608;&#xAD6C;&#xC5D0; &#xB300; &#xD55C; &#xC911;&#xC694; &#xD558;&#xB2E4;.

Modificaci&oacute;n de peque&ntilde;os ubiquitin-como modificador (SUMO) de prote&iacute;nas (sumoilaci&oacute;n) y deSUMOylation han surgido como importantes mecanismos de regulaci&oacute;n para la funci&oacute;n de la prote&iacute;na. SENP1 (proteasa SUMO espec&iacute;ficos) deconjugates SUMOs de prote&iacute;nas modificadas. Hemos creado ratones de nocaut (KO) SENP1 basados en un sistema Cre-loxP. Eliminaci&oacute;n global de SENP1 (SENP1 KO) causa anemia y mortalidad embrionaria entre el d&iacute;a embrionario 13.5 y d&iacute;a postnatal 1, correlacionando con defectos de la eritropoyesis en el h&iacute;gado fetal. Trasplante de m&eacute;dula de c&eacute;lulas del h&iacute;gado fetales SENP1 KO a recipientes adultos irradiados confiere eritropoyesis defectos. An&aacute;lisis de prote&iacute;na muestran que los genes GATA1 y GATA1 dependiente se regula en el h&iacute;gado fetal de ratones KO SENP1. Esta regulaci&oacute;n se correlaciona con la acumulaci&oacute;n de una forma de sumoiladas de GATA1. Adem&aacute;s, mostramos que SENP1 puede directamente deSUMOylate GATA1, regulaci&oacute;n de expresi&oacute;n g&eacute;nica de GATA1-dependiente y eritropoyesis ensayos in vitro. Por otra parte, demostramos que GATA1 sumoilaci&oacute;n altera su uni&oacute;n de ADN, reduciendo su contrataci&oacute;n para el promotor del gen GATA1-sensible. Colectivamente, concluimos que SENP1 promueve la activaci&oacute;n de GATA1 y eritropoyesis posterior por deSUMOylating GATA1.

Mediada por SENP1 GATA1 deSUMOylation es fundamental para la eritropoyesis definitiva.

ASK1-interacting protein-1 (AIP1), Ras GTPase-activating protein membre d&#39;une famille, est fortement exprim&eacute;e dans les cellules endoth&eacute;liales et musccells lisse vasculaire (CMLV). Le r&ocirc;le de AIP1 dans les CMLV et CMLV maladie prolif&eacute;rative ne conna&icirc;t pas.

AIP1 emp&ecirc;che le greffon art&eacute;rioscl&eacute;rose en inhibant la prolif&eacute;ration des cellules musculaires lisses de l&#39;interf&eacute;ron-&gamma;-d&eacute;pendante et expansion intimale.

ASK1-interazione proteina-1 (AIP1), un Ras Gap proteina membro della famiglia, &egrave; altamente espresso in cellule endoteliali e musccells lisce vascolari (VSMCs). Il ruolo di AIP1 in VSMCs e VSMC malattia proliferativa non &egrave; noto.

AIP1 impedisce l&#39;innesto arteriosclerosi inibendo la proliferazione delle cellule muscolari lisce di interferone-&gamma;-dipendente ed espansione intimale.

ASK1-interacting protein-1 (AIP1), a Ras GTPase-activating protein family member, is highly expressed in endothelial cells and vascular smooth musccells (VSMCs). The role of AIP1 in VSMCs and VSMC proliferative disease is not known.

AIP1 prevents graft arteriosclerosis by inhibiting interferon-Î³-dependent smooth muscle cell proliferation and intimal expansion.

Acetylcholinesterase (AChE) expression is pivotal during apoptosis. Indeed, AChE inhibitors partially protect cells from apoptosis. Insulin-dependent diabetes mellitus (IDDM) is characterized in part by pancreatic Î²-cell apoptosis. Here, we investigated the role of AChE in the development of IDDM and analyzed protective effects of AChE inhibitors. Multiple low-dose streptozotocin (MLD-STZ) administration resulted in IDDM in a mouse model. Western blot analysis, cytochemical staining, and immunofluorescence staining were used to detect AChE expression in MIN6 cells, primary Î² cells, and apoptotic pancreatic Î² cells of MLD-STZ-treated mice. AChE inhibitors were administered intraperitoneally to the MLD-STZ mice for 30 days. Blood glucose, plasma insulin, and creatine levels were measured, and glucose tolerance tests were performed. The effects of AChE inhibitors on MIN6 cells were also evaluated. AChE expression was induced in the apoptotic MIN6 cells and primary Î² cells in vitro and pancreatic islets in vivo when treated with STZ. Induction and progressive accumulation of AChE in the pancreatic islets were associated with apoptotic Î² cells during IDDM development. The administration of AChE inhibitors effectively decreased hyperglycemia and incidence of diabetes, and restored plasma insulin levels and plasma creatine clearance in the MLD-STZ mice. AChE inhibitors partially protected MIN6 cells from the damage caused by STZ treatment. Induction and accumulation of AChE in pancreatic islets and the protective effects of AChE inhibitors on the onset and development of IDDM indicate a close relationship between AChE and IDDM.

Acetylcholinesterase is associated with apoptosis in Î² cells and contributes to insulin-dependent diabetes mellitus pathogenesis.

Expression de l&#39;ac&eacute;tylcholinest&eacute;rase (AChE) est essentielle au cours de l&#39;apoptose. En effet, inhibiteurs de l&#39;AChE partiellement prot&egrave;gent les cellules de l&#39;apoptose. Mellitus de diab&egrave;te insulino - d&eacute;pendant (DID) se caract&eacute;rise en partie par apoptose des cellules &beta; pancr&eacute;atiques. Ici, nous avons &eacute;tudi&eacute; le r&ocirc;le de l&#39;AChE dans le d&eacute;veloppement du DSID et analys&eacute;es les effets protecteurs des inhibiteurs de la douleur. DID a entra&icirc;n&eacute; plusieurs administration de faibles doses de streptozotocine (STZ-MLD) dans un mod&egrave;le murin. Analyse par Western blot, coloration cytochimique et immunofluorescence souillant ont &eacute;t&eacute; utilis&eacute;s pour d&eacute;tecter l&#39;expression de l&#39;AChE dans MIN6 cellules, les cellules &beta; primaire et apoptotique des cellules &beta; pancr&eacute;atiques de souris trait&eacute;es au MLD-STZ. Inhibiteurs de la douleur ont &eacute;t&eacute; administr&eacute;s par voie intrap&eacute;riton&eacute;ale aux souris de MLD-STZ pendant 30 jours. On a mesur&eacute; les niveaux de cr&eacute;atine, insulin&eacute;mie et la glyc&eacute;mie, et les essais de tol&eacute;rance de glucose ont &eacute;t&eacute; effectu&eacute;es. Les effets des inhibiteurs de la douleur sur les cellules MIN6 ont &eacute;t&eacute; &eacute;galement &eacute;valu&eacute;s. Expression d&#39;estomac a &eacute;t&eacute; induite chez les cellules apoptotiques MIN6 et &beta; primaire des cellules in vitro et pancr&eacute;atiques &icirc;lots in vivo avec STZ. L&#39;induction et l&#39;accumulation progressive de l&#39;AChE dans les &icirc;lots pancr&eacute;atiques ont &eacute;t&eacute; associ&eacute;s &agrave; apoptotique des cellules &beta; au cours du d&eacute;veloppement de did. L&#39;administration d&#39;inhibiteurs de l&#39;AChE efficacement une diminution de l&#39;hyperglyc&eacute;mie et l&#39;incidence du diab&egrave;te et restaur&eacute; l&#39;insuline plasmatique et la clairance de cr&eacute;atine plasmatique chez les souris MLD-STZ. Inhibiteurs d&#39;estomac partiellement prot&eacute;g&eacute; MIN6 cellules contre les dommages caus&eacute;s par un traitement de STZ. Induction et accumulation d&#39;AChE en &icirc;lots pancr&eacute;atiques et les effets protecteurs des inhibiteurs de la douleur sur l&#39;apparition et le d&eacute;veloppement du DSID indiquent une relation &eacute;troite entre l&#39;estomac et did.

L&#39;ac&eacute;tylcholinest&eacute;rase est associ&eacute;e &agrave; l&#39;apoptose des cellules &beta; et contribue &agrave; la pathogen&egrave;se de mellitus de diab&egrave;te insulino - d&eacute;pendant.

Espressione dell&#39;acetilcolinesterasi (AChE) &egrave; fondamentale durante il apoptosis. Infatti, gli inibitori AChE parzialmente proteggono le cellule dall&#39;apoptosi. Mellito di diabete insulina - dipendente (IDDM) &egrave; caratterizzato in parte da &beta;-cellula pancreatica apoptosi. Qui, abbiamo studiato il ruolo della AChE in sviluppo di IDDM e analizzati effetti protettivi degli inibitori della AChE. Pi&ugrave; somministrazione di basse dosi streptozotocin (MLD-STZ) ha portato in IDDM in un modello murino. Analisi Western blot, colorazione citochimiche e colorazione di immunofluorescenza sono stati utilizzati per rilevare l&#39;espressione AChE in MIN6, cellule &beta; primario e cellule &beta; del pancreas apoptotic di topi MLD-STZ-trattati. Inibitori aChE sono stati somministrati per via intraperitoneale in topi MLD-STZ per 30 giorni. Glucosio nel sangue, plasma insulina e livelli di creatina sono stati misurati, e sono stati eseguiti test di tolleranza del glucosio. Inoltre sono stati valutati gli effetti degli inibitori della AChE su cellule MIN6. Mal di espressione &egrave; stata indotta in cellule apoptotic MIN6 e primaria &beta; cellule in vitro e del pancreas isolotti in vivo quando trattati con STZ. Induzione e progressivo accumulo di AChE negli isolotti pancreatici sono stati associati con apoptotic &beta; cellule durante lo sviluppo di IDDM. La somministrazione di inibitori della AChE effettivamente diminuito l&#39;iperglicemia e l&#39;incidenza del diabete e ripristinati i livelli di insulina nel plasma e plasma creatina clearance in topi MLD-STZ. Inibitori aChE parzialmente protetto MIN6 cellule dal danno causato da un trattamento STZ. Induzione e accumulo di dolore nelle isole pancreatiche e gli effetti protettivi degli inibitori AChE sull&#39;insorgenza e lo sviluppo di IDDM indicano una stretta relazione fra mal e IDDM.

Luyang Yu

Department of Pathology

Yale University School of Medicine

Mouse Models for Graft Arteriosclerosis

Luyang Yu

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

Yale University School of Medicine

Mouse Models for Graft Arteriosclerosis

Department of Pathology