a-multiplexed-luciferase-based-screening-platform-for-interrogating

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Genome-scale interrogation of gene function using RNA interference (RNAi)  holds tremendous promise for the rapid identification of chemically tractable  ...

UT Southwestern Medical Center

The Hedgehog (Hh) and Wnt signal transduction pathways are master regulators of embryogenesis and tissue renewal and represent anticancer therapeutic targets. Using genome-wide RNA interference screening in murine cultured cells, we established previously unknown associations between these signaling pathways and genes linked to developmental malformations, diseases of premature tissue degeneration, and cancer. We identified functions in both pathways for the multitasking kinase Stk11 (also known as Lkb1), a tumor suppressor implicated in lung and cervical cancers. We found that Stk11 loss resulted in disassembly of the primary cilium, a cellular organizing center for Hh pathway components, thus dampening Hh signaling. Loss of Stk11 also induced aberrant signaling through the Wnt pathway. Chemicals that targeted the Wnt acyltransferase Porcupine or that restored primary cilia length by inhibiting the tubulin deacetylase HDAC6 (histone deacetylase 6) countered deviant pathway activities driven by Stk11 loss. Our study demonstrates that Stk11 is a critical mediator in both the Hh and the Wnt pathways, and our approach provides a platform to support the development of targeted therapeutic strategies.

Genome-wide RNAi screen reveals disease-associated genes that are common to Hedgehog and Wnt signaling.

Le h&eacute;risson (Hh) et Wnt signalent voies de transduction sont des r&eacute;gulateurs ma&icirc;tres du renouveau de l&#39;embryogen&egrave;se et de tissus et repr&eacute;sentent des cibles th&eacute;rapeutiques anticanc&eacute;reuses. &Agrave; l&#39;aide de l&#39;interf&eacute;rence ARN G&eacute;nome-large de d&eacute;pistage dans des cellules cultiv&eacute;es, nous avons &eacute;tabli des associations in&eacute;dites entre ces voies de signalisation et des malformations li&eacute;es au d&eacute;veloppement de g&egrave;nes, des maladies de d&eacute;g&eacute;n&eacute;rescence des tissus pr&eacute;matur&eacute; et le cancer. Nous avons identifi&eacute; des fonctions dans les deux voies de la kinase de multit&acirc;che Stk11 (&eacute;galement connu sous le nom de Lkb1), un suppresseur de tumeur impliqu&eacute; dans le poumon et les cancers du col ut&eacute;rin. Nous avons trouv&eacute; que Stk11 perte entra&icirc;nerait d&eacute;montage du cil primaire, un t&eacute;l&eacute;phone cellulaire, Centre pour les composants de voie de Hh, organisateur amortissant ainsi la signalisation de Hh. Perte de Stk11 induit aussi aberrant de signalisation par la voie de signalisation Wnt. Produits chimiques qui cible l&#39;acyltransf&eacute;rase Wnt porc-&eacute;pic ou que restaur&eacute; longueur de cils primaires en inhibant les d&eacute;viants contr&eacute; de tubuline d&eacute;sac&eacute;tylases HDAC6 (histone deacetylase 6) activit&eacute;s de voie conduites par Stk11 perte. Notre &eacute;tude d&eacute;montre que Stk11 est un m&eacute;diateur critique dans le Hh et les voies de signalisation Wnt, et notre approche fournit une plate-forme pour soutenir le d&eacute;veloppement de strat&eacute;gies th&eacute;rapeutiques cibl&eacute;es.

&Eacute;cran de RNAi g&eacute;nome r&eacute;v&egrave;le les g&egrave;nes associ&eacute;s &agrave; la maladie qui sont communes &agrave; h&eacute;risson et Wnt signaling.

Il Hedgehog (Hh) e Wnt segnale vie di trasduzione del sono regolatori master di rinnovamento embriogenesi e tessuto e rappresentano bersagli terapeutici anticancro. Utilizzo di interferenza del RNA genome-wide screening in cellule coltivate murine, abbiamo stabilito precedentemente sconosciute associazioni tra queste vie di segnalazione e malformazioni legate alla sviluppo di geni, malattie di degenerazione precoce dei tessuti e cancro. Abbiamo individuato le funzioni in due percorsi per il multitasking chinasi Stk11 (noto anche come Lkb1), un soppressore tumorale implicato nel polmone e tumori della cervice uterina. Abbiamo trovato che Stk11 perdita ha portato a smontare il cilio primario, un cellulare organizzando il centro per i componenti del pathway di Hh, quindi inumidirsi Hh di segnalazione. Perdita di Stk11 indotta anche segnalazione aberrante attraverso il pathway Wnt. Sostanze chimiche che mirati il Wnt aciltransferasi istrice o che restaurata lunghezza ciliari inibendo deviante neutralizzata tubulina deacetylase HDAC6 (istone deacetylase 6) attivit&agrave; di percorso guidata da Stk11 perdita. Il nostro studio dimostra Stk11 &egrave; un mediatore critico sia l&#39;Hh e i percorsi di Wnt, che il nostro approccio fornisce una piattaforma per supportare lo sviluppo di strategie terapeutiche mirate.

Ozlem Kulak

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