the-ovo-chick-chorioallantoic-membrane-cam-assay-as-an-efficient

The <em>In Ovo </em>Chick Chorioallantoic Membrane (CAM) Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma

The chick chorioallantoic membrane (CAM) begins to develop by day 7 after fertilization and matures by day 12. The CAM is naturally immunodeficient and ...

Icahn School of Medicine at Mount Sinai

Breast cancers can recur after removal of the primary tumor and treatment to eliminate remaining tumor cells. Recurrence may occur after long periods of time during which there are no clinical symptoms. Tumor cell dormancy may explain these prolonged periods of asymptomatic residual disease and treatment resistance. We generated a dormancy gene signature from published experimental models and applied it to both breast cancer cell line expression data as well as four published clinical studies of primary breast cancers. We found that estrogen receptor (ER) positive breast cell lines and primary tumors have significantly higher dormancy signature scores (P<0.0000001) than ER- cell lines and tumors. In addition, a stratified analysis combining all ER+ tumors in four studies indicated 2.1 times higher hazard of recurrence among patients whose tumors had low dormancy scores (LDS) compared to those whose tumors had high dormancy scores (HDS) (p<0.000005). The trend was shown in all four individual studies. Suppression of two dormancy genes, BHLHE41 and NR2F1, resulted in increased in vivo growth of ER positive MCF7 cells. The patient data analysis suggests that disseminated ER positive tumor cells carrying a dormancy signature are more likely to undergo prolonged dormancy before resuming metastatic growth. Furthermore, genes identified with this approach might provide insight into the mechanisms of dormancy onset and maintenance as well as dormancy models using human breast cancer cell lines.

Dormancy signatures and metastasis in estrogen receptor positive and negative breast cancer.

2008 &#x5E74;&#x6211;&#x4EEC;&#x53D1;&#x8868;&#x4E86;&#x7B2C;&#x4E00;&#x5957;&#x6307;&#x5F15;&#xFF0C;&#x89C4;&#x8303;&#x81EA;&#x566C;&#x7684;&#x7814;&#x7A76;&#x3002;&#x81EA;&#x90A3;&#x65F6;&#x4EE5;&#x6765;&#xFF0C;&#x5173;&#x4E8E;&#x8FD9;&#x4E00;&#x4E3B;&#x9898;&#x7684;&#x7814;&#x7A76;&#x5DF2;&#x7EE7;&#x7EED;&#x52A0;&#x901F;&#xFF0C;&#x548C;&#x8BB8;&#x591A;&#x65B0;&#x79D1;&#x5B66;&#x5BB6;&#x5DF2;&#x8F93;&#x5165;&#x8BE5;&#x5B57;&#x6BB5;&#x3002;&#x6B64;&#x5916;&#x6269;&#x5927;&#x6211;&#x4EEC;&#x7684;&#x77E5;&#x8BC6;&#x5E93;&#x548C;&#x76F8;&#x5173;&#x65B0;&#x6280;&#x672F;&#x3002;&#x56E0;&#x6B64;&#xFF0C;&#x5B83;&#x662F;&#x91CD;&#x8981;&#x7684;&#x662F;&#x8981;&#x66F4;&#x65B0;&#x8FD9;&#x4E9B;&#x51C6;&#x5219;&#xFF0C;&#x4EE5;&#x4FBF;&#x76D1;&#x6D4B;&#x4E0D;&#x540C;&#x751F;&#x7269;&#x4F53;&#x4E2D;&#x7684;&#x81EA;&#x566C;&#x3002;&#x5404;&#x9879;&#x5BA1;&#x67E5;&#x6240;&#x63CF;&#x8FF0;&#x5DF2;&#x7528;&#x4E8E;&#x6B64;&#x76EE;&#x7684;&#x7684;&#x68C0;&#x6D4B;&#x7684;&#x8303;&#x56F4;&#x3002;&#x5C3D;&#x7BA1;&#x5982;&#x6B64;&#xFF0C;&#x7EE7;&#x7EED;&#x5173;&#x4E8E;&#x53EF;&#x63A5;&#x53D7;&#x7684;&#x65B9;&#x6CD5;&#x6765;&#x8861;&#x91CF;&#x81EA;&#x566C;&#xFF0C;&#x5C24;&#x5176;&#x662F;&#x5728;&#x591A;&#x7EC6;&#x80DE;&#x771F;&#x6838;&#x751F;&#x7269;&#x7684;&#x6DF7;&#x4E71;&#x3002;&#x9700;&#x8981;&#x5F3A;&#x8C03;&#x6307;&#x51FA;&#x7684;&#x5173;&#x952E;&#x70B9;&#x5C31;&#x662F;&#x76D1;&#x6D4B;&#x6570;&#x5B57;&#x6216;&#x5377;&#x7684;&#x81EA;&#x566C;&#x6027;&#x5143;&#x7D20; &#xFF08;&#x4F8B;&#x5982;&#xFF0C;autophagosomes &#x6216; autolysosomes&#xFF09; &#x7684;&#x6D4B;&#x91CF;&#x7ED3;&#x679C;&#x4E4B;&#x95F4;&#x7684;&#x5DEE;&#x5F02;&#x4E0E;&#x90A3;&#x4E9B;&#x81EA;&#x566C;&#x8FC7;&#x7A0B;&#x7684;&#x4EFB;&#x4F55;&#x9636;&#x6BB5;&#xFF0C;&#x68C0;&#x6D4B;&#x6D41;&#x91CF;&#x901A;&#x8FC7;&#x81EA;&#x566C;&#x9014;&#x5F84; &#xFF08;&#x5373;&#xFF0C;&#x5B8C;&#x6574;&#x7684;&#x8FDB;&#x7A0B;&#xFF09; &#xFF1B;&#x56E0;&#x6B64;&#xFF0C;macroautophagy autophagosome &#x79EF;&#x7D2F;&#x7684;&#x7ED3;&#x679C;&#x9700;&#x8981;&#x52A0;&#x4EE5;&#x533A;&#x5206;&#x4ECE;&#x81EA;&#x566C;&#x6027;&#x6D3B;&#x52A8;&#x589E;&#x52A0;&#xFF0C;&#x4F1A;&#x5BFC;&#x81F4;&#x7684;&#x523A;&#x6FC0;&#x4E2D;&#x7684;&#x5757;&#x5B9A;&#x4E49;&#x4E3A;&#x589E;&#x52A0;&#x81EA;&#x566C;&#x8BF1;&#x5BFC;&#x52A0;&#x4E0A;&#x589E;&#x52A0;&#x7684;&#x4EA4;&#x4ED8;&#x548C;&#x6EB6;&#x9176;&#x4F53; &#xFF08;&#x5728;&#x5927;&#x591A;&#x6570;&#x9AD8;&#x7B49;&#x771F;&#x6838;&#x751F;&#x7269;&#x548C;&#x76D8;&#x57FA;&#x7F51;&#x67C4;&#x7B49;&#x4E00;&#x4E9B;&#x751F;&#x7269;&#x4F53;&#xFF09; &#x6216; &#xFF08;&#x5728;&#x690D;&#x7269;&#x548C;&#x771F;&#x83CC;&#xFF09; &#x6CE1;&#x5185;&#x7684;&#x964D;&#x89E3;&#x3002;&#x6362;&#x53E5;&#x8BDD;&#x8BF4;&#xFF0C;&#x5B83;&#x662F;&#x5C24;&#x5176;&#x91CD;&#x8981;&#x7684;&#x662F;&#x65B0;&#x7684;&#x9886;&#x57DF;&#x7684;&#x8C03;&#x67E5;&#x4EBA;&#x5458;&#x4E86;&#x89E3;&#x66F4;&#x591A; autophagosomes &#x7684;&#x5916;&#x89C2;&#x4E0D;&#x4E00;&#x5B9A;&#x7B49;&#x540C;&#x66F4;&#x591A;&#x81EA;&#x566C;&#x3002;&#x4E8B;&#x5B9E;&#x4E0A;&#xFF0C;&#x5728;&#x8BB8;&#x591A;&#x60C5;&#x51B5;&#x4E0B;&#xFF0C;autophagosomes &#x79EF;&#x7D2F;&#x7531;&#x4E8E;&#x8D29;&#x8FD0;&#x5230;&#x6EB6;&#x9176;&#x4F53;&#x4E0D;&#x968F;&#x4E4B;&#x6539;&#x53D8;&#x7684;&#x751F;&#x7269;&#x53D1;&#x751F; autophagosome&#xFF0C;&#x4E00;&#x4E2A;&#x5757;&#x800C;&#x589E;&#x52A0; autolysosomes &#x53EF;&#x964D;&#x89E3;&#x6D3B;&#x52A8;&#x4E2D;&#x53CD;&#x6620;&#x51CF;&#x5C11;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x63D0;&#x51FA;&#x4E00;&#x5957;&#x51C6;&#x5219;&#x7684;&#x9009;&#x62E9;&#x548C;&#x4F7F;&#x7528;&#x65B9;&#x6CD5;&#x7684;&#x89E3;&#x91CA;&#x7531;&#x8C03;&#x67E5;&#x4EBA;&#x5458;&#xFF0C;&#x4ED6;&#x4EEC;&#x7684;&#x76EE;&#x7684;&#x662F;&#x68C0;&#x67E5; macroautophagy &#x548C;&#x76F8;&#x5173;&#x7684;&#x8FDB;&#x7A0B;&#xFF0C;&#x4EE5;&#x53CA;&#x5BA1;&#x9605;&#x8005;&#x9700;&#x8981;&#x63D0;&#x4F9B;&#x8BBA;&#x6587;&#x4FA7;&#x91CD;&#x4E8E;&#x8FD9;&#x4E9B;&#x8FDB;&#x7A0B;&#x7684;&#x73B0;&#x5B9E;&#x548C;&#x5408;&#x7406;&#x7684;&#x6279;&#x8BC4;&#x3002;&#x8FD9;&#x4E9B;&#x51C6;&#x5219;&#x4E0D;&#x662F;&#x8981;&#x5957;&#x516C;&#x5F0F;&#x5316;&#x7684;&#x89C4;&#x5219;&#xFF0C;&#x56E0;&#x4E3A;&#x9002;&#x5F53;&#x7684;&#x68C0;&#x6D4B;&#x90E8;&#x5206;&#x53D6;&#x51B3;&#x4E8E;&#x7684;&#x8D28;&#x8BE2;&#x548C;&#x6B63;&#x5728;&#x4F7F;&#x7528;&#x7684;&#x7CFB;&#x7EDF;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6211;&#x4EEC;&#x5F3A;&#x8C03;&#x4FDD;&#x8BC1;&#x6CA1;&#x6709;&#x4E2A;&#x522B;&#x6D4B;&#x5B9A;&#x90FD;&#x662F;&#x5728;&#x6BCF;&#x4E2A;&#x60C5;&#x51B5;&#x4E0B;&#xFF0C;&#x4E00;&#x4E2A;&#x6700;&#x5408;&#x9002;&#xFF0C;&#x6211;&#x4EEC;&#x5F3A;&#x70C8;&#x5EFA;&#x8BAE;&#x4F7F;&#x7528;&#x591A;&#x4E2A;&#x5316;&#x9A8C;&#x76D1;&#x6D4B;&#x81EA;&#x566C;&#x3002;&#x5728;&#x672C;&#x6307;&#x5357;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x8003;&#x8651;&#x8FD9;&#x4E9B;&#x4E0D;&#x540C;&#x7684;&#x65B9;&#x6CD5;&#xFF0C;&#x8BC4;&#x4F30;&#x81EA;&#x566C;&#x548C;&#x54EA;&#x4E9B;&#x4FE1;&#x606F;&#x53EF;&#x4EE5;&#xFF0C;&#x6216;&#x4E0D;&#x80FD;&#xFF0C;&#x4ECE;&#x5B83;&#x4EEC;&#x83B7;&#x5F97;&#x3002;&#x6700;&#x540E;&#xFF0C;&#x901A;&#x8FC7;&#x8BA8;&#x8BBA;&#x6848;&#x60C5;&#x548C;&#x7279;&#x5B9A;&#x7684;&#x81EA;&#x566C;&#x68C0;&#x6D4B;&#x65B9;&#x6CD5;&#x7684;&#x9650;&#x5236;&#xFF0C;&#x6211;&#x4EEC;&#x5E0C;&#x671B;&#x9F13;&#x52B1;&#x6280;&#x672F;&#x521B;&#x65B0;&#x7684;&#x9886;&#x57DF;&#x3002;

&#x68C0;&#x6D4B;&#x65B9;&#x6CD5;&#x76D1;&#x6D4B;&#x81EA;&#x566C;&#x7684;&#x89E3;&#x91CA;&#x548C;&#x4F7F;&#x7528;&#x7684;&#x6307;&#x5357;&#x3002;

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.

Guidelines for the use and interpretation of assays for monitoring autophagy.

En 2008, nous avons publi&eacute; le premier ensemble de lignes directrices pour la normalisation des recherches dans l&#39;autophagie. Depuis lors, les recherches sur ce sujet a continu&eacute; d&#39;acc&eacute;l&eacute;rer, et beaucoup de scientifiques nouveaux entr&eacute;s dans le champ. Notre base de connaissances et les nouvelles technologies ont &eacute;galement &eacute;tendu. En cons&eacute;quence, il est important de mettre &agrave; jour ces lignes directrices pour la surveillance de l&#39;autophagie dans diff&eacute;rents organismes. Diff&eacute;rents examens ont d&eacute;crit les divers essais qui ont &eacute;t&eacute; utilis&eacute;s &agrave; cet effet. N&eacute;anmoins, il continue d&#39;y avoir confusion au sujet des m&eacute;thodes acceptables pour mesurer l&#39;autophagie, surtout chez les eucaryotes multicellulaires. Un point essentiel qui doit &ecirc;tre soulign&eacute;, c&#39;est qu&#39;il y a une diff&eacute;rence entre les diff&eacute;rentes mesures qui surveillent les chiffres ou le volume des &eacute;l&eacute;ments autophagiques (autophagosomes ou autolysosomes) &agrave; tout stade du processus autophagique vs ceux ce flux de mesure par la voie de l&#39;autophagie (c.-&agrave;-d. le processus complet) ; ainsi, un bloc en macroautophagy, ce qui entra&icirc;ne l&#39;accumulation autophagosome doit &ecirc;tre diff&eacute;renci&eacute;s des stimuli qui r&eacute;sultent dans l&#39;augmentation de l&#39;activit&eacute; autophagique, d&eacute;finie comme augmentation autophagie induction coupl&eacute;e avec livraison accrue &agrave; et d&eacute;gradation dans les lysosomes (dans la plupart des eucaryotes sup&eacute;rieurs et certains protistes comme Dictyostelium) ou la vacuole (chez les plantes et les champignons). En d&#39;autres termes, il est particuli&egrave;rement important que les chercheurs &agrave; nouveau dans le domaine comprennent que l&#39;apparence des autophagosomes plus ne correspond pas n&eacute;cessairement avec autophagie plus. En effet, dans de nombreux cas, autophagosomes s&#39;accumulent en raison d&#39;un bloc dans le trafic vers les lysosomes sans changement concomitant autophagosome biogen&egrave;se, alors qu&#39;une augmentation d&#39;autolysosomes peut refl&eacute;ter une r&eacute;duction dans l&#39;activit&eacute; de d&eacute;gradation. Nous pr&eacute;sentons ici une s&eacute;rie de lignes directrices pour la s&eacute;lection et l&#39;interpr&eacute;tation des m&eacute;thodes pour l&#39;utilisation par les enqu&ecirc;teurs, qui visent &agrave; examiner les macroautophagy et les processus connexes, ainsi que pour les examinateurs qui doivent fournir des critiques r&eacute;alistes et raisonnables des papiers qui mettent l&#39;accent sur ces processus. Ces lignes directrices ne visent pas &agrave; &ecirc;tre un ensemble de formule de r&egrave;gles, parce que les dosages appropri&eacute;s d&eacute;pendent en partie de la question pos&eacute;e et le syst&egrave;me utilis&eacute;. En outre, nous soulignons qu&#39;aucun dosage individuel n&#39;est garanti d&#39;&ecirc;tre le plus appropri&eacute; dans chaque situation, et nous recommandons fortement l&#39;utilisation de multiples essais pour surveiller l&#39;autophagie. Dans ces lignes directrices, nous consid&eacute;rons ces diverses m&eacute;thodes d&#39;&eacute;valuation d&#39;autophagie et quels renseignements peuvent, ou ne peut &ecirc;tre obtenu d&#39;eux. Enfin, en discutant les avantages et les limites des analyses particuli&egrave;res autophagie, nous esp&eacute;rons encourager l&#39;innovation technique dans le domaine.

Lignes directrices pour l&#39;utilisation et l&#39;interpr&eacute;tation d&#39;essais pour la surveillance d&#39;autophagie.

Im Jahr 2008 haben wir den ersten Satz von Richtlinien f&uuml;r die Standardisierung der Forschung in Autophagy ver&ouml;ffentlicht. Seitdem Forschung zu diesem Thema hat sich weiter beschleunigen, und viele neue Wissenschaftler haben das Spielfeld betreten. Unsere Wissensdatenbank und relevante neue Technologien haben auch erweitert. Dementsprechend ist es wichtig, diese Leitlinien f&uuml;r die &Uuml;berwachung Autophagy in verschiedenen Organismen zu aktualisieren. Verschiedene Meinungen haben die Proben beschrieben, die f&uuml;r diesen Zweck verwendet wurden. Dennoch weiterhin besteht Verwirrung in Bezug auf akzeptierte, Autophagy, vor allem bei mehrzelligen Eukaryoten zu messen. Ein wesentlicher Punkt, der betont werden muss ist, dass es ein Unterschied zwischen Messungen, die &Uuml;berwachung der Zahlen oder Volume autophagic Elemente (z. B. Autophagosomes oder Autolysosomes) in jedem Stadium des Prozesses gegen die autophagic dieser Ma&szlig;nahme-Fluss durch den Autophagy Pathway (d. h. der vollst&auml;ndige Proze&szlig;); so ein Block in Macroautophagy, Ergebnisse in Autophagosomen Anh&auml;ufung von Reize unterschieden werden, die erh&ouml;hte autophagic Aktivit&auml;t, f&uuml;hren die definiert als erh&ouml;hte Autophagy Induktion gepaart mit erh&ouml;hter Lieferung an und Verschlechterung in Lysosomen (in den meisten h&ouml;heren Eukaryotes und einigen Einzellern wie Dictyostelium) oder die Vakuole (in Pflanzen und Pilzen). Mit anderen Worten, ist es besonders wichtig, dass die Ermittler in das Feld neue verstehen, dass die Darstellung des mehr Autophagosomes nicht unbedingt mit mehr Autophagy gleichzusetzen ist. In der Tat sammeln in vielen F&auml;llen Autophagosomes wegen einen Block in den Handel zu Lysosomen ohne eine damit einhergehende &Auml;nderung in Autophagosomen Biogenese, w&auml;hrend eine Zunahme der Autolysosomes eine Verringerung in Degradations-Aktivit&auml;t widerspiegeln kann. Hier pr&auml;sentieren wir eine Reihe von Leitlinien f&uuml;r die Auswahl und Interpretation der Methoden f&uuml;r die Verwendung von Ermittlern, die Macroautophagy und die damit verbundenen Prozesse untersuchen wollen, sowie f&uuml;r Pr&uuml;fer, die realistische und vern&uuml;nftige Kritiken von Papier bereitstellen, die auf diese Prozesse gerichtet werden m&uuml;ssen. Diese Richtlinien sollen kein formelhaften Satz von Regeln, weil die entsprechenden Assays teilweise auf die Frage und das System verwendet wird. Dar&uuml;ber hinaus betonen wir, dass keine einzelne Assay ist garantiert das passende Produkt in jeder Situation, und wir empfehlen die Verwendung von mehreren Proben Autophagy &uuml;berwachen. In diesen Leitlinien halten wir diese verschiedenen Methoden zur Bewertung Autophagy und welche Informationen kann, oder nicht, von ihnen abgerufen werden. Abschlie&szlig;end diskutieren die Vorz&uuml;ge und Grenzen von bestimmten Autophagy Assays, hoffen wir auf technische Innovationen im Bereich zu f&ouml;rdern.

Richtlinien f&uuml;r die Verwendung und Interpretation von Assays zur &Uuml;berwachung Autophagy in Verbindung.

Nel 2008 abbiamo pubblicato la prima serie di linee guida per la standardizzazione della ricerca in autophagy. Da allora, ricerca su questo argomento ha continuato ad accelerare, e molti nuovi scienziati hanno entrato in campo. Hanno anche ampliato la nostra knowledge base e le nuove tecnologie pertinenti. Di conseguenza, &egrave; importante aggiornare queste linee guida per il monitoraggio autophagy in diversi organismi. Varie recensioni hanno descritto la gamma di dosaggi che sono stati utilizzati per questo scopo. Tuttavia, ci continua a essere confusione per quanto riguarda i metodi accettabili per misurare autophagy, specialmente negli eucarioti pluricellulari. Un punto chiave che deve essere sottolineato &egrave; che c&#39;&egrave; una differenza tra le misure che monitorare il volume di autophagic elementi (ad esempio, autophagosomes o autolysosomes) o numeri in qualsiasi fase del processo autophagic vs quelli che misura flusso attraverso il pathway autophagy (cio&egrave;, il processo completo); cos&igrave;, un blocco in macroautophagy che risultati in accumulo di autophagosome deve essere differenziato da stimoli che si traducono in maggiore attivit&agrave; autophagic, definita come aumento autophagy induzione accoppiato con consegna aumentata e la degradazione nei lisosomi (nella maggior parte dei eucarioti superiori e alcuni protisti come Dictyostelium) o il vacuolo (in piante e funghi). In altre parole, &egrave; particolarmente importante che gli investigatori di nuovi al campo capiscano che l&#39;aspetto di pi&ugrave; autophagosomes non necessariamente identificare con pi&ugrave; autophagy. Infatti, in molti casi, autophagosomes si accumulano a causa di un blocco nel traffico ai lisosomi senza un cambiamento concomitante di autophagosome biogenesi, considerando che un aumento di autolysosomes pu&ograve; riflettere una riduzione nell&#39;attivit&agrave; degradanti. Qui, vi presentiamo una serie di linee guida per la selezione e l&#39;interpretazione dei metodi per l&#39;uso dagli investigatori che mirano a esaminare macroautophagy e dei processi correlati, nonch&eacute; per i revisori che hanno bisogno di fornire critiche realistiche e ragionevole delle carte che sono focalizzate su questi processi. Queste linee guida non sono destinate ad essere una serie stereotipata di regole, perch&eacute; i dosaggi appropriati dipendono in parte la domanda e il sistema in uso. Inoltre, sottolineiamo che nessun dosaggio individuale &egrave; garantito per essere quello pi&ugrave; appropriato in ogni situazione, e si consiglia vivamente l&#39;uso di dosaggi multipli di monitorare autophagy. In queste linee guida, consideriamo questi vari metodi di valutazione autophagy e quali informazioni possono, o non pu&ograve; essere ottenuto da loro. Infine, da discutere i meriti e i limiti delle analisi particolare autophagy, speriamo di incoraggiare l&#39;innovazione tecnica nel campo.

Linee guida per l&#39;uso e l&#39;interpretazione delle analisi per il monitoraggio autophagy.

2008 &#x5E74;&#x306B;&#x6700;&#x521D;&#x306E;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x306E;&#x7814;&#x7A76;&#x6A19;&#x6E96;&#x5316;&#x306E;&#x305F;&#x3081;&#x306E;&#x30AC;&#x30A4;&#x30C9;&#x30E9;&#x30A4;&#x30F3;&#x306E;&#x30BB;&#x30C3;&#x30C8;&#x3092;&#x516C;&#x958B;&#x3057;&#x307E;&#x3059;&#x3002;&#x305D;&#x306E;&#x5F8C;&#x3001;&#x3053;&#x306E;&#x30C8;&#x30D4;&#x30C3;&#x30AF;&#x306B;&#x95A2;&#x3059;&#x308B;&#x7814;&#x7A76;&#x3092;&#x52A0;&#x901F;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;&#x7D9A;&#x3051;&#x3066;&#x3044;&#x308B;&#x3001;&#x591A;&#x304F;&#x306E;&#x65B0;&#x3057;&#x3044;&#x79D1;&#x5B66;&#x8005;&#x53C2;&#x5165;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x79C1;&#x9054;&#x306E;&#x77E5;&#x8B58;&#x30D9;&#x30FC;&#x30B9;&#x3068;&#x95A2;&#x9023;&#x3059;&#x308B;&#x65B0;&#x3057;&#x3044;&#x30C6;&#x30AF;&#x30CE;&#x30ED;&#x30B8;&#x3082;&#x62E1;&#x5927;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;&#x7570;&#x306A;&#x308B;&#x751F;&#x7269;&#x306B;&#x304A;&#x3051;&#x308B;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x3092;&#x76E3;&#x8996;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3053;&#x308C;&#x3089;&#x306E;&#x30AC;&#x30A4;&#x30C9;&#x30E9;&#x30A4;&#x30F3;&#x3092;&#x66F4;&#x65B0;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x91CD;&#x8981;&#x3067;&#x3059;&#x3002;&#x69D8;&#x3005; &#x306A;&#x30EC;&#x30D3;&#x30E5;&#x30FC;&#x306F;&#x3053;&#x306E;&#x76EE;&#x7684;&#x306E;&#x305F;&#x3081;&#x306B;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x306E;&#x7BC4;&#x56F2;&#x306B;&#x3064;&#x3044;&#x3066;&#x8AAC;&#x660E;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x305D;&#x308C;&#x306B;&#x3082;&#x304B;&#x304B;&#x308F;&#x3089;&#x305A;&#x3001;&#x7279;&#x306B;&#x591A;&#x7D30;&#x80DE;&#x771F;&#x6838;&#x751F;&#x7269;&#x306B;&#x304A;&#x3051;&#x308B;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x3092;&#x6E2C;&#x5B9A;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x53D7;&#x3051;&#x5165;&#x308C;&#x65B9;&#x6CD5;&#x306B;&#x95A2;&#x3059;&#x308B;&#x6DF7;&#x4E71;&#x7D9A;&#x3051;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x91CD;&#x8981;&#x306A;&#x30DD;&#x30A4;&#x30F3;&#x30C8;&#x3092;&#x5F37;&#x8ABF;&#x3059;&#x308B;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x756A;&#x53F7;&#x307E;&#x305F;&#x306F;&#x30DC;&#x30EA;&#x30E5;&#x30FC;&#x30E0; autophagic &#x8981;&#x7D20; &#xFF08;&#x4F8B;&#x3048;&#x3070;&#x3001;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B4;&#x30BD;&#x30FC;&#x30E0;&#x307E;&#x305F;&#x306F; autolysosomes) &#x306E;&#x76E3;&#x8996;&#x6E2C;&#x5B9A;&#x306E;&#x9055;&#x3044;&#x3082;&#x306E;&#x5BFE; autophagic &#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306E;&#x3069;&#x306E;&#x6BB5;&#x968E;&#x3067;&#x3082;&#x305D;&#x306E;&#x30E1;&#x30B8;&#x30E3;&#x30FC; &#x30D5;&#x30E9;&#x30C3;&#x30AF;&#x30B9; &#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x7D4C;&#x8DEF; (&#x3059;&#x306A;&#x308F;&#x3061;&#x3001;&#x5B8C;&#x4E86;&#x306E;&#x30D7;&#x30ED;&#x30BB;&#x30B9;); &#x3092;&#x4ECB;&#x3057;&#x3066;&#x3067;&#x3059;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B4;&#x30BD;&#x30FC;&#x30E0;&#x84C4;&#x7A4D;&#x306E;&#x7D50;&#x679C;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308B;&#x5897;&#x52A0;&#x306E; autophagic &#x6D3B;&#x52D5;&#x306E;&#x7D50;&#x679C;&#x523A;&#x6FC0;&#x304B;&#x3089;&#x533A;&#x5225;&#x3055;&#x308C;&#x308B;&#x305F;&#x3081;&#x306B;&#x3067;&#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x3092;&#x5B9A;&#x7FA9;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;&#x5897;&#x52A0;&#x306E;&#x914D;&#x4FE1;&#x3068;&#x52A3;&#x5316;&#x30EA;&#x30BD;&#x30BD;&#x30FC;&#x30E0; &#xFF08;&#x306E;&#x307B;&#x3068;&#x3093;&#x3069;&#x9AD8;&#x7B49;&#x771F;&#x6838;&#x751F;&#x7269;&#x3068;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E;&#x539F;&#x751F;&#x751F;&#x7269;&#x7D30;&#x80DE;&#x6027;&#x7C98;&#x83CC;&#x306A;&#x3069;&#xFF09; &#x307E;&#x305F;&#x306F; &#xFF08;&#x306E;&#x690D;&#x7269;&#x3068;&#x83CC;&#x985E;&#xFF09; &#x6DB2;&#x80DE;&#x5185;&#x7D50;&#x5408;&#x5897;&#x52A0;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x8A98;&#x5C0E;&#x3068;&#x3057;&#x3066;&#x3002;&#x4ED6;&#x306E;&#x8A00;&#x8449;&#x3067;&#x306F;&#x3001;&#x635C;&#x67FB;&#x5B98;&#x306F;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x306B;&#x65B0;&#x3057;&#x3044;&#x8A73;&#x7D30;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B4;&#x30BD;&#x30FC;&#x30E0;&#x306E;&#x5916;&#x89B3;&#x306F;&#x3001;&#x5FC5;&#x305A;&#x3057;&#x3082;&#x3088;&#x308A;&#x591A;&#x304F;&#x306E;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x3068;&#x3092;&#x4E00;&#x81F4;&#x3057;&#x306A;&#x3044;&#x3092;&#x7406;&#x89E3;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x7279;&#x306B;&#x91CD;&#x8981;&#x3067;&#x3059;&#x3002;&#x5B9F;&#x969B;&#x306B;&#x306F;&#x3001;autolysosomes &#x306E;&#x5897;&#x52A0;&#x5206;&#x89E3;&#x6D3B;&#x6027;&#x306E;&#x6E1B;&#x5C11;&#x3092;&#x53CD;&#x6620;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x306B;&#x5BFE;&#x3057;&#x30EA;&#x30BD;&#x30BD;&#x30FC;&#x30E0;&#x4F75;&#x7528;&#x5909;&#x5316;&#x306A;&#x3057;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B4;&#x30BD;&#x30FC;&#x30E0;&#x5F62;&#x6210;&#x306B;&#x304A;&#x3051;&#x308B;&#x4EBA;&#x8EAB;&#x58F2;&#x8CB7;&#x3067;&#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x306E;&#x305F;&#x3081;&#x591A;&#x304F;&#x306E;&#x5834;&#x5408;&#x304C;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B4;&#x30BD;&#x30FC;&#x30E0;&#x84C4;&#x7A4D;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x4E00;&#x9023;&#x306E;&#x9078;&#x629E;&#x306E;&#x305F;&#x3081;&#x306E;&#x30AC;&#x30A4;&#x30C9;&#x30E9;&#x30A4;&#x30F3;&#x3068;&#x5229;&#x7528;&#x65B9;&#x6CD5;&#x306E;&#x89E3;&#x91C8;&#x3068;&#x95A2;&#x9023;&#x3059;&#x308B;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x3092;&#x691C;&#x8A0E;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x76EE;&#x6307;&#x3057;&#x3066;&#x8ABF;&#x67FB;&#x5B98;&#x306B;&#x3088;&#x3063;&#x3066;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x3001;&#x6821;&#x95B2;&#x8005;&#x304C;&#x3053;&#x308C;&#x3089;&#x306E;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306B;&#x7126;&#x70B9;&#x3092;&#x5F53;&#x3066;&#x3066;&#x3044;&#x308B;&#x66F8;&#x985E;&#x306E;&#x73FE;&#x5B9F;&#x7684;&#x304B;&#x3064;&#x5408;&#x7406;&#x7684;&#x306A;&#x6279;&#x5224;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308B;&#x305F;&#x3081;&#x672C;.&#x9069;&#x5207;&#x306A;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x306E;&#x4E00;&#x90E8;&#x3001;&#x8CEA;&#x554F;&#x3068;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x3044;&#x308B;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306B;&#x4F9D;&#x5B58;&#x3057;&#x3066;&#x3044;&#x308B;&#x305F;&#x3081;&#x306B;&#x5B9A;&#x578B;&#x7684;&#x306A;&#x4E00;&#x9023;&#x306E;&#x30EB;&#x30FC;&#x30EB;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x30AC;&#x30A4;&#x30C9;&#x30E9;&#x30A4;&#x30F3;&#x306F;&#x610F;&#x5473;&#x3055;&#x308C;&#x307E;&#x305B;&#x3093;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x3059;&#x3079;&#x3066;&#x306E;&#x72B6;&#x6CC1;&#x3067;&#x6700;&#x3082;&#x9069;&#x5207;&#x306A; 1 &#x3064;&#x306B;&#x500B;&#x3005; &#x306E;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x306F;&#x4FDD;&#x8A3C;&#x3055;&#x308C;&#x307E;&#x305B;&#x3093;&#x304A;&#x3088;&#x3073;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x3092;&#x76E3;&#x8996;&#x3059;&#x308B;&#x8907;&#x6570;&#x306E;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x306E;&#x4F7F;&#x7528;&#x3092;&#x304A;&#x52E7;&#x3081;&#x3092;&#x5F37;&#x8ABF;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x30AC;&#x30A4;&#x30C9;&#x30E9;&#x30A4;&#x30F3;&#x3067;&#x306F;&#x3001;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x3068;&#x3069;&#x306E;&#x3088;&#x3046;&#x306A;&#x60C5;&#x5831;&#x3092;&#x3067;&#x304D;&#x308B;&#x3001;&#x8A55;&#x4FA1;&#x306E;&#x3053;&#x308C;&#x3089;&#x306E;&#x3055;&#x307E;&#x3056;&#x307E;&#x306A;&#x65B9;&#x6CD5;&#x3092;&#x691C;&#x8A0E;&#x3057;&#x3066;&#x304F;&#x3060;&#x3055;&#x3044;&#x307E;&#x305F;&#x306F;&#x305D;&#x308C;&#x3089;&#x304B;&#x3089;&#x53D6;&#x5F97;&#x3059;&#x308B;&#x3053;&#x3068;&#x306F;&#x3067;&#x304D;&#x307E;&#x305B;&#x3093;&#x3002;&#x6700;&#x5F8C;&#x306B;&#x3001;&#x30E1;&#x30EA;&#x30C3;&#x30C8;&#x3068;&#x7279;&#x5B9A;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x306E;&#x8A66;&#x91D1;&#x306E;&#x9650;&#x754C;&#x3092;&#x8AD6;&#x8B70;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x3063;&#x3066;&#x3001;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x3067;&#x306E;&#x6280;&#x8853;&#x9769;&#x65B0;&#x3092;&#x5968;&#x52B1;&#x3059;&#x308B;&#x9858;&#x3063;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#x4F7F;&#x7528;&#x3068;&#x89E3;&#x91C8;&#x30AA;&#x30FC;&#x30C8;&#x30D5;&#x30A1;&#x30B8;&#x30FC;&#x3092;&#x76E3;&#x8996;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x8A66;&#x91D1;&#x306E;&#x305F;&#x3081;&#x306E;&#x30AC;&#x30A4;&#x30C9;&#x30E9;&#x30A4;&#x30F3;&#x3002;

2008 &#xB144; &#xC6B0;&#xB9AC; &#xCD9C;&#xD310; &#xC5F0;&#xAD6C; autophagy&#xC758; &#xD45C;&#xC900;&#xD654;&#xC5D0; &#xB300; &#xD55C; &#xC9C0;&#xCE68;&#xC758; &#xCCAB; &#xC138;&#xD2B8;. &#xADF8; &#xC774;&#xD6C4;&#xB85C;, &#xAC00;&#xC18D;,&#xC774; &#xC8FC;&#xC81C;&#xC5D0; &#xB300; &#xD55C; &#xC5F0;&#xAD6C;&#xB294; &#xACC4;&#xC18D; &#xD558; &#xACE0; &#xB9CE;&#xC740; &#xC0C8;&#xB85C;&#xC6B4; &#xACFC;&#xD559;&#xC790; &#xD544;&#xB4DC;&#xB97C; &#xC785;&#xB825; &#xD588;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xAE30;&#xC220; &#xC790;&#xB8CC; &#xBC0F; &#xAD00;&#xB828; &#xC2E0;&#xAE30;&#xC220;&#xB3C4; &#xD655;&#xB300; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC774; &#xB530;&#xB77C;, &#xADF8;&#xAC83;&#xC740; &#xB2E4;&#xB978; &#xC720;&#xAE30; &#xCCB4;&#xC5D0;&#xC11C; autophagy &#xBAA8;&#xB2C8;&#xD130;&#xB9C1;&#xC5D0; &#xB300; &#xD55C;&#xC774; &#xC9C0;&#xCE68;&#xC744; &#xC5C5;&#xB370;&#xC774;&#xD2B8; &#xD558;&#xB294; &#xAC83;&#xC774; &#xC911;&#xC694;&#xC785;&#xB2C8;&#xB2E4;. &#xB2E4;&#xC591; &#xD55C; &#xB9AC;&#xBDF0;&#xB294;&#xC774; &#xBAA9;&#xC801;&#xC744; &#xC704;&#xD574; &#xC0AC;&#xC6A9; &#xB41C; &#xBD84;&#xC11D; &#xC2E4;&#xD5D8;&#xC758; &#xBC94;&#xC704;&#xB97C; &#xC124;&#xBA85; &#xD588;&#xB2E4;. &#xADF8;&#xB7FC;&#xC5D0;&#xB3C4; &#xBD88;&#xAD6C; &#xD558; &#xACE0;, &#xAC70;&#xAE30;&#xC5D0; &#xD5C8;&#xC6A9; &#xD2B9;&#xD788; &#xB2E4;&#xC138;&#xD3EC; &#xC9C4;&#xD575;&#xC0DD;&#xBB3C;&#xC5D0;&#xC11C;&#xC5D0;&#xC11C; autophagy &#xCE21;&#xC815; &#xBC29;&#xBC95;&#xC5D0; &#xAD00;&#xD55C; &#xD63C;&#xB780; &#xB418; &#xACE0; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xAC15;&#xC870; &#xD558;&#xB294; &#xD575;&#xC2EC; &#xD3EC;&#xC778;&#xD2B8;&#xAC00;&#xC785;&#xB2C8;&#xB2E4; &#xC22B;&#xC790; &#xB610;&#xB294; autophagic &#xC694;&#xC18C; (&#xC608;: autophagosomes &#xB610;&#xB294; autolysosomes)&#xC758; &#xBCFC;&#xB968;&#xC744; &#xBAA8;&#xB2C8;&#xD130;&#xB9C1; &#xD558;&#xB294; &#xCE21;&#xC815;&#xC758; &#xCC28;&#xC774; &#xADF8; &#xB300; autophagic &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC758; &#xBAA8;&#xB4E0; &#xB2E8;&#xACC4;&#xC5D0;&#xC11C; autophagy &#xD1B5;&#xB85C; (&#xC989;, &#xC804;&#xCCB4; &#xD504;&#xB85C;&#xC138;&#xC2A4;);&#xB97C; &#xD1B5;&#xD574; &#xADF8; &#xCE21;&#xC815; &#xC720;&#xB7C9; &#xB530;&#xB77C;&#xC11C;, &#xACB0;&#xACFC;: autophagosome &#xCD95;&#xC801; &#xC99D;&#xAC00; autophagic &#xD65C;&#xB3D9;&#xC5D0;&#xC11C; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xC790;&#xADF9;&#xC5D0;&#xC11C; &#xCC28;&#xBCC4;&#xD654; &#xD558;&#xB294; macroautophagy&#xC5D0;&#xC11C; &#xBE14;&#xB85D; &#xC99D;&#xAC00; autophagy &#xC720;&#xB3C4;, &#xC99D;&#xAC00; &#xC804;&#xB2EC;&#xACFC; &#xB9AC;&#xC18C;&#xC880; (&#xB300;&#xBD80;&#xBD84;&#xC758; &#xB192;&#xC740; &#xC9C4;&#xD575;&#xC0DD;&#xBB3C;&#xC5D0; Dictyostelium &#xAC19;&#xC740; &#xC77C;&#xBD80; protists) &#xB610;&#xB294; (&#xC2DD;&#xBB3C;&#xACFC; &#xACF0; &#xD321;&#xC774;)&#xC5D0; &#xACF5;&#xD3EC; &#xB0B4; &#xC800;&#xD558;&#xC640; &#xACB0;&#xD569;&#xC73C;&#xB85C; &#xC815;&#xC758; &#xB429;&#xB2C8;&#xB2E4;. &#xB2E4;&#xB978; &#xB9D0;&#xB85C; &#xD558;&#xBA74;, &#xADF8;&#xAC83;&#xC740; &#xD2B9;&#xD788; &#xC911;&#xC694; &#xD55C; &#xC218;&#xC0AC; &#xBD84;&#xC57C;&#xC5D0; &#xC0C8;&#xB85C;&#xC6B4; &#xB354; &#xB9CE;&#xC740; autophagosomes&#xC758; &#xC678;&#xAD00;&#xC744; &#xBC18;&#xB4DC;&#xC2DC; &#xB354; autophagy&#xC640; &#xB3D9;&#xC77C;&#xC2DC; &#xD558;&#xC9C0; &#xC54A;&#xB294;&#xB2E4; &#xC774;&#xD574;. &#xC0AC;&#xC2E4;, &#xB300;&#xBD80;&#xBD84;&#xC758; &#xACBD;&#xC6B0;&#xC5D0;&#xC11C; autophagosomes &#xCD95;&#xC801; autophagosome &#xC18D;&#xC5D0; &#xC218; &#xBC18;&#xD558;&#xB294; &#xBCC0;&#xD654; &#xC5C6;&#xC774; &#xB9AC;&#xC18C;&#xC880;&#xC744; &#xC778;&#xC2E0; &#xB9E4;&#xB9E4;&#xC5D0; &#xBE14;&#xB85D; &#xB54C;&#xBB38;&#xC5D0; &#xBC18;&#xBA74;&#xC5D0; autolysosomes &#xC99D;&#xAC00; degradative &#xD65C;&#xB3D9;&#xC758; &#xAC10;&#xC18C;&#xB97C; &#xBC18;&#xC601; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xAC00; &#xD604;&#xC7AC; &#xC120;&#xD0DD; &#xC601;&#xC5ED;&#xC5D0; &#xB300; &#xD55C; &#xC9C0;&#xCE68;&#xC758; &#xC124;&#xC815; &#xBC0F; &#xC0AC;&#xC6A9; &#xBC29;&#xBC95;&#xC758; &#xD574;&#xC11D; &#xC774;&#xB7EC;&#xD55C; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC5D0; &#xC9D1;&#xC911; &#xB418;&#xB294; &#xB17C;&#xBB38;&#xC758; &#xD604;&#xC2E4;&#xC801;&#xC774; &#xACE0; &#xD569;&#xB9AC;&#xC801;&#xC778; &#xBE44;&#xD310;&#xC744; &#xC81C;&#xACF5; &#xD574;&#xC57C; &#xD558;&#xB294; &#xAC80;&#xD1A0;&#xC790;&#xC5D0; &#xB300; &#xD55C; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; macroautophagy &#xBC0F; &#xAD00;&#xB828;&#xB41C; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xB97C; &#xAC80;&#xC0AC; &#xD558;&#xB294; &#xAC83;&#xC744; &#xBAA9;&#xD45C;&#xB85C; &#xD558;&#xB294; &#xC870;&#xC0AC;&#xC5D0; &#xC758;&#xD574;. &#xC801;&#xC808; &#xD55C; &#xBD84;&#xC11D; &#xC2E4;&#xD5D8; &#xBD80;&#xBD84;&#xC5D0;&#xC11C; &#xC694;&#xCCAD;&#xC744; &#xBC1B;&#xACE0; &#xC9C8;&#xBB38;&#xC744; &#xD558; &#xACE0; &#xC0AC;&#xC6A9; &#xC911;&#xC778; &#xC2DC;&#xC2A4;&#xD15C;&#xC5D0; &#xC758;&#xC874; &#xD558;&#xAE30; &#xB54C;&#xBB38;&#xC5D0; &#xC774;&#xB7EC;&#xD55C; &#xC9C0;&#xCE68; &#xADDC;&#xCE59;, formulaic &#xC9D1;&#xD569;&#xC73C;&#xB85C; &#xC758;&#xBBF8;&#xAC00; &#xC5C6;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xC6B0;&#xB9AC;&#xB294; &#xC544;&#xBB34; &#xAC1C;&#xBCC4; &#xBD84;&#xC11D; &#xACB0;&#xACFC;, &#xBAA8;&#xB4E0; &#xC0C1;&#xD669;&#xC5D0;&#xC11C; &#xAC00;&#xC7A5; &#xC801;&#xD569; &#xD55C; &#xBCF4;&#xC7A5; &#xD558; &#xACE0; &#xC88B;&#xC2B5;&#xB2C8;&#xB2E4; autophagy &#xBAA8;&#xB2C8;&#xD130;&#xB9C1;&#xD560; &#xC5EC;&#xB7EC; &#xBD84;&#xC11D; &#xC2E4;&#xD5D8;&#xC758; &#xC0AC;&#xC6A9;&#xC744; &#xAC15;&#xC870;. &#xC774;&#xB7EC;&#xD55C; &#xC9C0;&#xCE68;&#xC5D0; &#xC6B0;&#xB9AC; autophagy&#xC640; &#xC815;&#xBCF4; &#xC218; &#xC788;&#xB294; &#xD3C9;&#xAC00;&#xC758; &#xC774;&#xB7EC;&#xD55C; &#xB2E4;&#xC591; &#xD55C; &#xBC29;&#xBC95;&#xC744; &#xACE0;&#xB824; &#xADF8;&#xB4E4; &#xB85C;&#xBD80;&#xD130; &#xC5BB;&#xC744; &#xC218; &#xC5C6;&#xC2B5;&#xB2C8;&#xB2E4;, &#xB610;&#xB294;. &#xB9C8;&#xC9C0;&#xB9C9;&#xC73C;&#xB85C;, &#xC7A5;&#xC810; &#xBC0F; &#xD2B9;&#xC815; autophagy &#xBD84;&#xC11D; &#xC2E4;&#xD5D8;&#xC758; &#xD55C;&#xACC4;&#xB97C; &#xB17C;&#xC758; &#xD558; &#xC5EC; &#xD604;&#xC7A5;&#xC5D0;&#xC11C; &#xAE30;&#xC220; &#xD601;&#xC2E0;&#xC744; &#xC7A5;&#xB824; &#xD558;&#xACA0;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xC0AC;&#xC6A9; &#xBC0F; autophagy &#xBAA8;&#xB2C8;&#xD130;&#xB9C1; &#xBD84;&#xC11D; &#xC2E4;&#xD5D8;&#xC758; &#xD574;&#xC11D;&#xC5D0; &#xB300; &#xD55C; &#xC9C0;&#xCE68;.

En 2008 public&oacute; el primer conjunto de pautas para estandarizar la investigaci&oacute;n en la autofagia. Desde entonces, ha continuado la investigaci&oacute;n sobre este tema acelerar, y muchos nuevos cient&iacute;ficos han entrado en el campo. Tambi&eacute;n han estado ampliando nuestra base de conocimientos y tecnolog&iacute;as relevantes. Por consiguiente, es importante actualizar estas directrices para la supervisi&oacute;n de autofagia en diferentes organismos. Varios comentarios han descrito la gama de ensayos que se han utilizado para este prop&oacute;sito. Sin embargo, contin&uacute;an confusi&oacute;n con respecto a los m&eacute;todos aceptables para medir la autofagia, especialmente en eucariotas pluricelulares. Un punto clave que debe destacarse es que hay una diferencia entre las medidas que monitorean los n&uacute;meros o volumen de autofagia elementos (por ejemplo, autophagosomes o autolysosomes) en cualquier etapa del proceso de autofagia vs los ese flujo medida a trav&eacute;s de la v&iacute;a de la autofagia (es decir, el proceso completo); as&iacute;, un bloque en macroautophagy que resulta en acumulaci&oacute;n de autophagosome debe ser distinguido de est&iacute;mulos que resultan en un aumento de la actividad autofagia, definido como inducci&oacute;n autofagia creciente juntada con mayor entrega y degradaci&oacute;n dentro, lisosomas (en la mayor&iacute;a de eucariotas superiores y algunos protistas como Dictyostelium) o la vacuola (en plantas y hongos). En otras palabras, es especialmente importante que los investigadores de nuevos al campo entiendan que la aparici&oacute;n de m&aacute;s autophagosomes no corresponde necesariamente con autofagia m&aacute;s. De hecho, en muchos casos, autophagosomes se acumulan debido a un bloque en el tr&aacute;fico a los lisosomas sin un cambio concomitante en la biog&eacute;nesis de autophagosome, mientras que un aumento en la autolysosomes puede reflejar una reducci&oacute;n en la actividad de degradaci&oacute;n. Aqu&iacute;, presentamos un conjunto de directrices para la selecci&oacute;n e interpretaci&oacute;n de los m&eacute;todos para su uso por los investigadores que tienen como objetivo examinar macroautophagy y procesos relacionados, as&iacute; como para los revisores que deban proporcionar cr&iacute;ticas realistas y razonables de papeles que se centran en estos procesos. Estas directrices no pretenden ser un formulista conjunto de reglas, porque los ensayos adecuados dependen en parte de la pregunta y el sistema utilizado. Adem&aacute;s, hacemos hincapi&eacute; en que ning&uacute;n ensayo individual est&aacute; garantizada para ser el m&aacute;s adecuado en cada situaci&oacute;n, y le recomendamos el uso de m&uacute;ltiples ensayos para monitorear la autofagia. En estas directrices, consideramos estos diversos m&eacute;todos de evaluaci&oacute;n de autofagia y qu&eacute; informaci&oacute;n puede, o no puede obtenerse de ellos. Finalmente, discutiendo los m&eacute;ritos y los l&iacute;mites de an&aacute;lisis particular de la autofagia, queremos fomentar la innovaci&oacute;n t&eacute;cnica en el campo.

Directrices para el uso e interpretaci&oacute;n de ensayos para el control de la autofagia.

The development of metastasis is the major cause of death in cancer patients. In certain instances, this occurs shortly after primary tumor detection and treatment, indicating these lesions were already expanding at the moment of diagnosis or initiated exponential growth shortly after. However, in many types of cancer, patients succumb to metastatic disease years and sometimes decades after being treated for a primary tumor. This has led to the notion that in these patients residual disease may remain in a dormant state. Tumor cell dormancy is a poorly understood phase of cancer progression and only recently have its underlying molecular mechanisms started to be revealed. Important questions that remain to be elucidated include not only which mechanisms prevent residual disease from proliferating but also which mechanisms critically maintain the long-term survival of these disseminated residual cells. Herein, we review recent evidence in support of genetic and epigenetic mechanisms driving dormancy. We also explore how therapy may cause the onset of dormancy in the surviving fraction of cells after treatment and how autophagy may be a mechanism that maintains the residual cells that are viable for prolonged periods.

Regulation of tumor cell dormancy by tissue microenvironments and autophagy.

Emerging evidence points to aberrant regulation of translation as a driver of cell transformation in cancer. Given the direct control of translation by tRNA modifications, tRNA modifying enzymes may function as regulators of cancer progression. Here, we show that a tRNA methyltransferase 9-like (hTRM9L/KIAA1456) mRNA is down-regulated in breast, bladder, colorectal, cervix and testicular carcinomas. In the aggressive SW620 and HCT116 colon carcinoma cell lines, hTRM9L is silenced and its re-expression and methyltransferase activity dramatically suppressed tumour growth in vivo. This growth inhibition was linked to decreased proliferation, senescence-like G0/G1-arrest and up-regulation of the RB interacting protein LIN9. Additionally, SW620 cells re-expressing hTRM9L did not respond to hypoxia via HIF1-α-dependent induction of GLUT1. Importantly, hTRM9L-negative tumours were highly sensitive to aminoglycoside antibiotics and this was associated with altered tRNA modification levels compared to antibiotic resistant hTRM9L-expressing SW620 cells. Our study links hTRM9L and tRNA modifications to inhibition of tumour growth via LIN9 and HIF1-α-dependent mechanisms. It also suggests that aminoglycoside antibiotics may be useful to treat hTRM9L-deficient tumours.

A human tRNA methyltransferase 9-like protein prevents tumour growth by regulating LIN9 and HIF1-α.

Metastasis awakening: targeting dormant cancer.

In patients, non-proliferative disseminated tumour cells (DTCs) can persist in the bone marrow (BM) while other organs (such as lung) present growing metastasis. This suggested that the BM might be a metastasis 'restrictive soil' by encoding dormancy-inducing cues in DTCs. Here we show in a head and neck squamous cell carcinoma (HNSCC) model that strong and specific transforming growth factor-β2 (TGF-β2) signalling in the BM activates the MAPK p38α/β, inducing an (ERK/p38)(low) signalling ratio. This results in induction of DEC2/SHARP1 and p27, downregulation of cyclin-dependent kinase 4 (CDK4) and dormancy of malignant DTCs. TGF-β2-induced dormancy required TGF-β receptor-I (TGF-β-RI), TGF-β-RIII and SMAD1/5 activation to induce p27. In lungs, a metastasis 'permissive soil' with low TGF-β2 levels, DTC dormancy was short-lived and followed by metastatic growth. Importantly, systemic inhibition of TGF-β-RI or p38α/β activities awakened dormant DTCs, fuelling multi-organ metastasis. Our work reveals a 'seed and soil' mechanism where TGF-β2 and TGF-β-RIII signalling through p38α/β regulates DTC dormancy and defines restrictive (BM) and permissive (lung) microenvironments for HNSCC metastasis.

TGF-β2 dictates disseminated tumour cell fate in target organs through TGF-β-RIII and p38α/β signalling.

Integration of microenvironmental and stress signaling antagonizes colorectal cancer progression.

Metastases arise from residual disseminated tumour cells (DTCs). This can happen years after primary tumour treatment because residual tumour cells can enter dormancy and evade therapies. As the biology of minimal residual disease seems to diverge from that of proliferative lesions, understanding the underpinnings of this new cancer biology is key to prevent metastasis. Analysis of approximately 7 years of literature reveals a growing focus on tumour and normal stem cell quiescence, extracellular and stromal microenvironments, autophagy and epigenetics as mechanisms that dictate tumour cell dormancy. In this Review, we attempt to integrate this information and highlight both the weaknesses and the strengths in the field to provide a framework to understand and target this crucial step in cancer progression.

Mechanisms of disseminated cancer cell dormancy: an awakening field.

Cancer dormancy refers to the prolonged clinical disease-free time between removal of the primary tumor and recurrence, which is common in prostate cancer (PCa), breast cancer, esophageal cancer, and other cancers. PCa disseminated tumor cells (DTC) are detected in both patients with no evidence of disease (NED) and advanced disease (ADV). However, the molecular and cellular nature of DTC is unknown. We performed a first-in-field study of single DTC transcriptomic analyses in cancer patients to identify a molecular signature associated with cancer dormancy. We profiled eighty-five individual EpCAM⁺/CD45⁻ cells from the bone marrow of PCa patients with NED or ADV. We analyzed 44 DTC with high prostate-epithelial signatures, and eliminated 41 cells with high erythroid signatures and low prostate epithelial signatures. DTC were clustered into 3 groups: NED, ADV_1, and ADV_2, in which the ADV_1 group presented a distinct gene expression pattern associated with the p38 stress activated kinase pathway. Additionally, DTC from the NED group were enriched for a tumor dormancy signature associated with head and neck squamous carcinoma and breast cancer. This study provides the first clinical evidence of the p38 pathway as a potential biomarker for early recurrence and an attractive target for therapeutic intervention.

Characterization of single disseminated prostate cancer cells reveals tumor cell heterogeneity and identifies dormancy associated pathways.

Metastases can originate from disseminated tumour cells (DTCs), which may be dormant for years before reactivation. Here we find that the orphan nuclear receptor NR2F1 is epigenetically upregulated in experimental head and neck squamous cell carcinoma (HNSCC) dormancy models and in DTCs from prostate cancer patients carrying dormant disease for 7-18 years. NR2F1-dependent dormancy is recapitulated by a co-treatment with the DNA-demethylating agent 5-Aza-C and retinoic acid across various cancer types. NR2F1-induced quiescence is dependent on SOX9, RARβ and CDK inhibitors. Intriguingly, NR2F1 induces global chromatin repression and the pluripotency gene NANOG, which contributes to dormancy of DTCs in the bone marrow. When NR2F1 is blocked in vivo, growth arrest or survival of dormant DTCs is interrupted in different organs. We conclude that NR2F1 is a critical node in dormancy induction and maintenance by integrating epigenetic programmes of quiescence and survival in DTCs.

NR2F1 controls tumour cell dormancy via SOX9- and RARβ-driven quiescence programmes.

Xbp1, a key mediator of the unfolded protein response (UPR), is activated by IRE1α-mediated splicing, which results in a frameshift to encode a protein with transcriptional activity. However, the direct function of Xbp1 in epithelial cells during mammary gland development is unknown. Here we report that the loss of Xbp1 in the mammary epithelium through targeted deletion leads to poor branching morphogenesis, impaired terminal end bud formation, and spontaneous stromal fibrosis during the adult virgin period. Additionally, epithelial Xbp1 deletion induces endoplasmic reticulum (ER) stress in the epithelium and dramatically inhibits epithelial proliferation and differentiation during lactation. The synthesis of milk and its major components, α/β-casein and whey acidic protein (WAP), is significantly reduced due to decreased prolactin receptor (Prlr) and ErbB4 expression in Xbp1-deficient mammary epithelium. Reduction of Prlr and ErbB4 expression and their diminished availability at the cell surface lead to reduced phosphorylated Stat5, an essential regulator of cell proliferation and differentiation during lactation. As a result, lactating mammary glands in these mice produce less milk protein, leading to poor pup growth and postnatal death. These findings suggest that the loss of Xbp1 induces a terminal UPR which blocks proliferation and differentiation during mammary gland development.

Epithelial Xbp1 is required for cellular proliferation and differentiation during mammary gland development.

The mechanisms allowing residual multiple myeloma (MM) cells to persist after bortezomib (Bz) treatment remain unclear. We hypothesized that studying the biology of bortezomib-surviving cells may reveal markers to identify these cells and survival signals to target and kill residual MM cells.

Article	Year
Dormancy signatures and metastasis in estrogen receptor positive and negative breast cancer. PloS one\| PubMed ID: 22530051	2012
Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy\| PubMed ID: 22966490	2012
Regulation of tumor cell dormancy by tissue microenvironments and autophagy. Advances in experimental medicine and biology\| PubMed ID: 23143976	2013
A human tRNA methyltransferase 9-like protein prevents tumour growth by regulating LIN9 and HIF1-α. EMBO molecular medicine\| PubMed ID: 23381944	2013
Metastasis awakening: targeting dormant cancer. Nature medicine\| PubMed ID: 23467238	2013
TGF-β2 dictates disseminated tumour cell fate in target organs through TGF-β-RIII and p38α/β signalling. Nature cell biology\| PubMed ID: 24161934	2013
Integration of microenvironmental and stress signaling antagonizes colorectal cancer progression. The EMBO journal\| PubMed ID: 25024435	2014
Mechanisms of disseminated cancer cell dormancy: an awakening field. Nature reviews. Cancer\| PubMed ID: 25118602	2014
Characterization of single disseminated prostate cancer cells reveals tumor cell heterogeneity and identifies dormancy associated pathways. Oncotarget\| PubMed ID: 25301725	2014
NR2F1 controls tumour cell dormancy via SOX9- and RARβ-driven quiescence programmes. Nature communications\| PubMed ID: 25636082	2015
Epithelial Xbp1 is required for cellular proliferation and differentiation during mammary gland development. Molecular and cellular biology\| PubMed ID: 25713103	2015
Identification of markers that functionally define a quiescent multiple myeloma cell sub-population surviving bortezomib treatment. BMC cancer\| PubMed ID: 26025442	2015
Origin and interpretation of cancer transcriptome profiling: the essential role of the stroma in determining prognosis and drug resistance. EMBO molecular medicine\| PubMed ID: 26240076	2015

Julio A. Aguirre-Ghiso

Biography

JoVE Journal Publications

Other Publications