live-cell-imaging-assays-to-study-glioblastoma-brain-tumor-stem-cell

Live-Cell Imaging Assays to Study Glioblastoma Brain Tumor Stem Cell Migration and Invasion

Glioblastoma (GBM) is an aggressive brain tumor that is poorly controlled with the currently available treatment options. Key features of GBMs include ...

University of Calgary

The Hospital for Sick Children

&#x72EC;&#x7279;&#x7684;&#x4E34;&#x5E8A;&#x3001; &#x75C5;&#x7406;&#x3001; &#x548C;&#x9057;&#x4F20;&#x529F;&#x80FD;&#x7684;&#x7279;&#x70B9;&#x662F;&#x9274;&#x522B;&#x3002;&#x67D3;&#x8272;&#x4F53; 1 p &#x548C; 19q &#x7ECF;&#x5E38;&#x6027;&#x635F;&#x5931;&#x5F3A;&#x70C8;&#x5173;&#x8054;&#x6B64;&#x8111;&#x80BF;&#x7624;&#x4F46;&#x77E5;&#x8BC6;&#x7684;&#x8EAB;&#x4EFD;&#x548C;&#x53D7;&#x8FD9;&#x4E9B;&#x53D8;&#x5316;&#x7684;&#x57FA;&#x56E0;&#x7684;&#x4F5C;&#x7528;&#x4E5F;&#x5F88;&#x6709;&#x9650;&#x3002;&#x6211;&#x4EEC;&#x5728; 16 oligodendrogliomas 1 p/19q co-deletion&#xFF0C;&#x4EE5;&#x786E;&#x5B9A;&#x5728;&#x78B1;&#x57FA;&#x5BF9;&#x51B3;&#x8BAE;&#x7684;&#x65B0;&#x5206;&#x5B50;&#x529F;&#x80FD;&#x4E0E;&#x53D1;&#x73B0;&#x96C6;&#x4E0A;&#x6267;&#x884C; exome &#x6D4B;&#x5E8F;&#x3002;&#x6B63;&#x5982;&#x9884;&#x671F;&#xFF0C;&#x6709;&#x7684; IDH &#x7A81;&#x53D8;&#x7387;&#x5F88;&#x9AD8;&#xFF1A; &#x6240;&#x6709;&#x6848;&#x4EF6;&#x90FD;&#x5728;&#x4EFB;&#x4E00; IDH1 &#x7A81;&#x53D8; (14/16&#xFF09; &#x6216; IDH2 &#xFF08;2/16&#xFF09;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x4F53;&#x7EC6;&#x80DE;&#x7A81;&#x53D8;&#x548C;&#x63D2;&#x5165;/&#x5220;&#x9664;&#x5728;&#x4E2D;&#x6295;&#x516C;&#x53F8;&#x57FA;&#x56E0;&#x5728;&#x67D3;&#x8272;&#x4F53; 19q13.2 13/16 &#x80BF;&#x7624;&#x4E2D;&#x3002;&#x7A81;&#x53D8;&#xFF0C;&#x8BA4;&#x53EF;&#x4E86;&#x6DF1;&#x6D4B;&#x5E8F;&#x7684; 13 &#x989D;&#x5916;&#x7684;&#x80BF;&#x7624;&#xFF0C;&#x4E03;&#x4EBA;&#x4E0E;&#x4E2D;&#x6295;&#x516C;&#x53F8;&#x7684;&#x7A81;&#x53D8;&#xFF0C;&#x4ECE;&#x800C;&#x63ED;&#x793A;&#x8FD9;&#x4E9B;&#x53D1;&#x73B0;&#x5957; oligodendrogliomas &#x5728;&#x8FD9;&#x9879;&#x7814;&#x7A76;&#x5728;&#x63D0;&#x8BF7;&#x603B;&#x4F53;&#x53D8;&#x5F02;&#x7387; 20/29 &#xFF08;69%&#xFF09;&#x3002;&#x76F8;&#x6BD4;&#x4E4B;&#x4E0B;&#xFF0C;&#x661F;&#x5F62;&#x7EC6;&#x80DE;&#x548C;&#x65E0; 1 p/19q &#x635F;&#x5931; oligoastrocytomas &#x6DF1;&#x6D4B;&#x5E8F;&#x8868;&#x660E;&#x4E2D;&#x6295;&#x516C;&#x53F8;&#x6539;&#x5EFA;&#x5426;&#x5219;&#x7F55;&#x89C1; (1/60 &#xFF1B; 2%)&#x3002;21 &#x975E;&#x540C;&#x4E49;&#x4F53;&#x7EC6;&#x80DE;&#x7A81;&#x53D8;&#x7684; 20 &#x4E2D;&#x6295;&#x7A81;&#x53D8;&#x4F53; oligodendrogliomas&#xFF0C;&#x4E5D;&#x6B63;&#x5728;&#x5916;&#x663E;&#x5B50; 5 &#x9644;&#x52A0;&#x8BF4;&#x660E; DNA &#x4EA4;&#x4E92;&#x57DF;&#x5185;&#xFF0C;&#x4E09;&#x4E2A;&#x5916;&#x663E;&#x5B50; 20 &#x9644;&#x52A0;&#x8BF4;&#x660E;&#x86CB;&#x767D;&#x4EA4;&#x4E92;&#x57DF;&#x5185;&#x3002;&#x5176;&#x4F59;&#x4E5D;&#x540D;&#x88AB;&#x53D1;&#x73B0;&#x5728;&#x5176;&#x4ED6;&#x7528;&#xFF0C;&#x7ECF;&#x5E38;&#x5305;&#x62EC;&#x622A;&#x65AD;&#x3002;&#x4E2D;&#x6295;&#x516C;&#x53F8;&#x7684;&#x7A81;&#x53D8;&#x90FD;&#x4E0E;&#x9274;&#x522B;&#x7EC4;&#x7EC7;&#x5B66;&#x3001; 1 p/19q co-deletion &#x548C; IDH1/2 &#x7A81;&#x53D8; (p &lt; 0.001&#xFF09; &#x9AD8;&#x5EA6;&#x5173;&#x8054;&#x3002;&#x867D;&#x7136;&#x6211;&#x4EEC;&#x89C2;&#x5BDF;&#x5230;&#x4E34;&#x5E8A;&#x6210;&#x679C;&#x4E2D;&#x6295;&#x4E0E;&#x91CE;&#x751F;&#x578B;&#x80BF;&#x7624;&#xFF0C;1 p/19q co-deletion &#x7684;&#x80CC;&#x666F;&#x4E2D;&#x7684;&#x7A81;&#x53D8;&#x4F53;&#x4E2D;&#x6CA1;&#x6709;&#x5DEE;&#x5F02; hemizygous &#x4E2D;&#x6295;&#x7A81;&#x53D8;&#x5F88;&#x53EF;&#x80FD;&#x91CD;&#x8981;&#x3002;&#x6211;&#x4EEC;&#x63A8;&#x6D4B;&#x4E0A;&#x7684;&#x5355;&#x4E2A;&#x4FDD;&#x7559;&#x7684; 19q &#x7B49;&#x4F4D;&#x57FA;&#x56E0;&#x7684;&#x7A81;&#x53D8;&#x4E2D;&#x6295;&#x94FE;&#x63A5;&#x5230; oligodendrogliomas &#x4E0E; IDH &#x7A81;&#x53D8;&#x7684;&#x53D1;&#x75C5;&#x673A;&#x5236;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x9274;&#x522B;&#x4E2D;&#x7684;&#x9057;&#x4F20;&#x53D8;&#x5F02;&#x7684;&#x8BE6;&#x7EC6;&#x7684;&#x7814;&#x7A76;&#x8868;&#x660E;&#x529F;&#x80FD;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x4E2D;&#x6295;&#x7A81;&#x53D8;&#xFF0C;&#x7A81;&#x53D8; IDH1/2 &#x4E0E; 1 p/19q co-deletion&#x3002;

&#x5E76;&#x53D1;&#x4E2D;&#x6295;&#x7A81;&#x53D8;&#x3001; IDH &#x7A81;&#x53D8;&#x548C; 1 p/19q &#x635F;&#x5931;&#x533A;&#x5206; oligodendrogliomas &#x5176;&#x4ED6;&#x764C;&#x75C7;&#x3002;

Oligodendroglioma is characterized by unique clinical, pathological, and genetic features. Recurrent losses of chromosomes 1p and 19q are strongly associated with this brain cancer but knowledge of the identity and function of the genes affected by these alterations is limited. We performed exome sequencing on a discovery set of 16 oligodendrogliomas with 1p/19q co-deletion to identify new molecular features at base-pair resolution. As anticipated, there was a high rate of IDH mutations: all cases had mutations in either IDH1 (14/16) or IDH2 (2/16). In addition, we discovered somatic mutations and insertions/deletions in the CIC gene on chromosome 19q13.2 in 13/16 tumours. These discovery set mutations were validated by deep sequencing of 13 additional tumours, which revealed seven others with CIC mutations, thus bringing the overall mutation rate in oligodendrogliomas in this study to 20/29 (69%). In contrast, deep sequencing of astrocytomas and oligoastrocytomas without 1p/19q loss revealed that CIC alterations were otherwise rare (1/60; 2%). Of the 21 non-synonymous somatic mutations in 20 CIC-mutant oligodendrogliomas, nine were in exon 5 within an annotated DNA-interacting domain and three were in exon 20 within an annotated protein-interacting domain. The remaining nine were found in other exons and frequently included truncations. CIC mutations were highly associated with oligodendroglioma histology, 1p/19q co-deletion, and IDH1/2 mutation (p < 0.001). Although we observed no differences in the clinical outcomes of CIC mutant versus wild-type tumours, in a background of 1p/19q co-deletion, hemizygous CIC mutations are likely important. We hypothesize that the mutant CIC on the single retained 19q allele is linked to the pathogenesis of oligodendrogliomas with IDH mutation. Our detailed study of genetic aberrations in oligodendroglioma suggests a functional interaction between CIC mutation, IDH1/2 mutation, and 1p/19q co-deletion.

Concurrent CIC mutations, IDH mutations, and 1p/19q loss distinguish oligodendrogliomas from other cancers.

Oligodendrogliome se caract&eacute;rise par des caract&eacute;ristiques uniques de cliniques, pathologiques et g&eacute;n&eacute;tiques. Pertes courantes de chromosomes 1 p et 19 q sont fortement associ&eacute;es &agrave; ce cancer du cerveau, mais la connaissance de l&#39;identit&eacute; et la fonction des g&egrave;nes touch&eacute;s par ces modifications sont limit&eacute;es. Nous avons r&eacute;alis&eacute; le s&eacute;quen&ccedil;age de l&#39;exome sur un jeu de d&eacute;couverte de 16 oligodendrogliomes avec 1 p/19 q co-deletion pour identifier les nouveaux dispositifs mol&eacute;culaires &agrave; r&eacute;solution de paires de bases. Comme pr&eacute;vu, il y avait un taux &eacute;lev&eacute; de mutations de l&#39;IDH : tous les cas pr&eacute;sentaient des mutations en soit IDH1 (14/16) ou IDH2 (2/16). En outre, nous avons d&eacute;couvert que des mutations somatiques et insertions/d&eacute;l&eacute;tions dans le g&egrave;ne CIC sur le chromosome 19 tumeurs de 13/16. Ces jeu de d&eacute;couverte des mutations ont &eacute;t&eacute; valid&eacute;es par le s&eacute;quen&ccedil;age en profondeur de 13 tumeurs suppl&eacute;mentaires, sept autres personnes pr&eacute;sentant des mutations de CIC, r&eacute;v&eacute;lant ainsi portant le taux global de mutation oligodendrogliomes dans cette &eacute;tude &agrave; 20/29 (69 %). En revanche, s&eacute;quen&ccedil;age en profondeur des astrocytomes et oligoastrocytomas sans perte de 19 p/q 1 a r&eacute;v&eacute;l&eacute; que les alt&eacute;rations de CIC &eacute;taient autres rare (1/60; 2 %). Des mutations somatiques non-synonymes 21 dans 20 CIC-mutant oligodendrogliomes, neuf &eacute;taient dans l&#39;exon 5 dans un domaine d&#39;interaction ADN annot&eacute; et trois dans l&#39;exon 20 dans un domaine d&#39;interaction prot&eacute;ine annot&eacute;. Les neuf autres ont &eacute;t&eacute; retrouv&eacute;s dans les autres exons et fr&eacute;quemment inclus troncations. Des mutations de CIC ont &eacute;t&eacute; fortement associ&eacute;es &agrave; l&#39;histologie oligodendrogliome, 1 19 p/q co-deletion et mutation IDH1/2 (p &lt; 0,001). Bien que nous avons n&#39;observ&eacute; aucune diff&eacute;rence dans les r&eacute;sultats cliniques de mutant CIC contre des tumeurs de type sauvage, dans un contexte de co-deletion 1 p/19 q, h&eacute;mizygote CIC mutations sont probables importantes. Nous &eacute;mettons l&#39;hypoth&egrave;se que le mutant CIC sur l&#39;all&egrave;le unique 19 retenu q est li&eacute; &agrave; la pathog&eacute;nie des oligodendrogliomes avec la mutation de l&#39;IDH. Notre &eacute;tude d&eacute;taill&eacute;e des aberrations g&eacute;n&eacute;tiques chez Oligodendrogliome sugg&egrave;re une interaction fonctionnelle entre la mutation du CIC, mutation IDH1/2 et 1 19 p/q co-deletion.

Mutations simultan&eacute;es de CIC, mutations de l&#39;IDH et 1 perte de 19 p/q distinguent oligodendrogliomes d&#39;autres cancers.

Oligodendrogliom zeichnet sich durch einzigartige klinische und pathologische genetische Eigenschaften. Wiederkehrende Verluste von Chromosomen 1 p und 19q, nicht distale sind eng verbunden mit diesem Gehirntumoren, aber beschr&auml;nkt sich Kenntnisse &uuml;ber die Identit&auml;t und Funktion der Gene, die von diesen &Auml;nderungen betroffen. Wir spielten Exome Sequenzierung auf einem Entdeckung 16 Oligodendrogliome mit 1 p/19q, nicht distale Co-deletion Molekulare Neuerungen bei Basenpaar Aufl&ouml;sung zu identifizieren. Wie erwartet, gab es eine hohe Rate von IDH Mutationen: allen F&auml;llen hatten Mutationen in beiden IDH1 (14/16) oder IDH2 (2/16). Dar&uuml;ber hinaus entdeckten wir somatische Mutationen und Einf&uuml;gungen/L&ouml;schungen in der CIC-gen auf Chromosom 19q13.2 13/16 Tumoren. Diese Entdeckung-Reihe, die Mutationen durch Ultratiefe Sequenzierung von 13 zus&auml;tzliche Tumoren, validiert wurden die sieben andere CIC Mutationen, so offenbart bringen das Gesamtniveau der Mutation in Oligodendrogliome in dieser Studie an 20/29 (69 %). Im Gegensatz dazu Ultratiefe Sequenzierung von Astrozytome und Oligoastrocytomas ohne 1 p/19q, nicht distale Verlust ergab, dass CIC &Auml;nderungen sonst selten (1/60, 2 %). Von 21 nicht Synonym somatischen Mutationen in 20 CIC-Mutant Oligodendrogliome neun waren in Exon 5 innerhalb einer kommentierten DNA-Interaktion-Dom&auml;ne und drei waren in Exon 20 in einer kommentierten-Protein-Interaktion-Dom&auml;ne. Die restlichen neun fanden sich in anderen Exons und enthalten h&auml;ufig K&uuml;rzungen. CIC-Mutationen wurden hoch mit Oligodendrogliom Histologie, 1 p/19q, nicht distale Co-deletion und IDH1/2-Mutation (p &lt; 0,001). Obwohl wir keine Unterschiede in der klinischen Resultate der CIC Mutant im Vergleich zu Wildtyp Tumoren, bei dem Hintergrund 1 p/19q, nicht distale Co-deletion, beobachtet Hemizygous CIC Mutationen werden wahrscheinlich wichtig. Wir vermuten, dass die Pathogenese der Oligodendrogliome mit IDH Mutation der Mutant CIC auf die einzelne beibehaltenen 19q, nicht distale Allel verkn&uuml;pft ist. Unsere detaillierte Studie &uuml;ber genetische Aberrationen in Oligodendrogliom schl&auml;gt eine funktionelle Interaktion zwischen CIC Mutation, Mutation IDH1/2 und 1 p/19q, nicht distale Co-deletion.

Gleichzeitige CIC-Mutationen, IDH-Mutationen und 1 p/19q, nicht distale Verlust unterscheiden Oligodendrogliome von anderen Krebsarten.

Oligodendroglioma &egrave; caratterizzato da caratteristiche cliniche, patologiche e genetiche uniche. Perdite ricorrenti dei cromosomi 1 p e 19q sono fortemente associate a questo tipo di tumore cerebrale, ma conoscenza dell&#39;identit&agrave; e la funzione dei geni interessati da queste alterazioni &egrave; limitati. Abbiamo eseguito exome sequenziamento su un set di scoperta di 16 oligodendrogliomi con 1 p/19q co-deletion per identificare nuove caratteristiche molecolari alla base-coppia risoluzione. Come anticipato, c&#39;era un alto tasso di mutazioni IDH: tutti i casi presentavano mutazioni in entrambi IDH1 (14/16) o IDH2 (2/16). Inoltre, abbiamo scoperto le mutazioni somatiche e inserzioni/delezioni del gene CIC sul cromosoma 19q13.2 nei tumori 13/16. Questi set di scoperta le mutazioni sono state Validate dal profondo sequenziamento di 13 tumori supplementare, che ha rivelato altri sette con mutazioni del CIC, cos&igrave; portando il tasso di mutazione globale in oligodendrogliomi in questo studio a 20/29 (69%). Al contrario, deep sequencing di astrocitomi e oligoastrocitoma senza 1 p/19q perdita ha rivelato che alterazioni CIC furono altrimenti rara (1/60; 2%). 21 Non sinonimo somatico mutazioni in 20 CIC-mutante oligodendrogliomi, nove erano nell&#39;esone 5 all&#39;interno di un dominio di interazione del DNA con annotazioni e tre erano nell&#39;esone 20 all&#39;interno di un dominio di interazione della proteina con annotazioni. I restanti nove sono stati trovati in altri esoni e frequentemente incluso troncamenti. Mutazioni CIC sono stati altamente associate con oligodendroglioma istologia, 1 co-deletion p/19q e mutazione IDH1/2 (p &lt; 0.001). Anche se abbiamo non osservato alcuna differenza nei risultati clinici del mutante CIC contro tumori wild-type, in uno sfondo di co-deletion 1 p/19q, emizigoti CIC mutazioni sono probabilmente importanti. Ipotizziamo che il mutante CIC su 19q mantenuto singolo allele &egrave; legato alla patogenesi della oligodendrogliomi con mutazione IDH. Il nostro studio dettagliato delle aberrazioni genetiche in oligodendroglioma suggerisce un&#39;interazione funzionale tra mutazione CIC, mutazione IDH1/2 e 1 co-deletion p/19q.

Mutazioni simultanee di CIC, mutazioni IDH e 1 p/19q perdita distinguere gli oligodendrogliomi da altri tumori.

&#x3053;&#x3046;&#x3057;&#x3085;&#x81E8;&#x5E8A;&#x75C5;&#x7406;&#x5B66;&#x7684;, &#x3068;&#x907A;&#x4F1D;&#x306E;&#x72EC;&#x81EA;&#x306E;&#x6A5F;&#x80FD;&#x306B;&#x3088;&#x3063;&#x3066;&#x7279;&#x5FB4;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x307E;&#x3059;&#x3002;&#x67D3;&#x8272;&#x4F53; 1 p &#x3068; 19q &#x306E;&#x518D;&#x767A;&#x306E;&#x640D;&#x5931;&#x304C;&#x3053;&#x306E;&#x8133;&#x764C;&#x3068;&#x5F37;&#x304F;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x5909;&#x5316;&#x306B;&#x3088;&#x3063;&#x3066;&#x5F71;&#x97FF;&#x3092;&#x53D7;&#x3051;&#x308B;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x6A5F;&#x80FD;&#x3068;&#x30A2;&#x30A4;&#x30C7;&#x30F3;&#x30C6;&#x30A3;&#x30C6;&#x30A3;&#x306E;&#x77E5;&#x8B58;&#x304C;&#x9650;&#x3089;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x30C7;&#x30A3;&#x30B9;&#x30AB;&#x30D0;&#x30EA;&#x30FC; &#x30BB;&#x30C3;&#x30C8; 16 oligodendroglioma &#x5869;&#x57FA;&#x5BFE;&#x306E;&#x89E3;&#x50CF;&#x5EA6;&#x3067;&#x65B0;&#x3057;&#x3044;&#x5206;&#x5B50;&#x6A5F;&#x80FD;&#x3092;&#x8B58;&#x5225;&#x3059;&#x308B; 1 p/19q co-deletion &#x3068;&#x306E;&#x30A8;&#x30AD;&#x30BD;&#x30FC;&#x30E0;&#x3092;&#x5B9F;&#x884C;&#x3057;&#x307E;&#x3059;&#x3002;&#x4E88;&#x60F3;&#x901A;&#x308A;&#x3001;IDH &#x5909;&#x7570;&#x306E;&#x7387;&#x304C;&#x9AD8;&#x3044;&#x304C;&#x3042;&#x3063;&#x305F;&#xFF1A; &#x3059;&#x3079;&#x3066;&#x306E;&#x30B1;&#x30FC;&#x30B9;&#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x304C;&#x3042;&#x3063;&#x305F;&#x3069;&#x3061;&#x3089;&#x304B; IDH1 &#x3067; (14/16) &#x307E;&#x305F;&#x306F; IDH2 (2/16)&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x4F53;&#x7D30;&#x80DE;&#x5909;&#x7570;&#x3068;&#x633F;&#x5165;/&#x524A;&#x9664; 13/16 &#x816B;&#x760D;&#x306B;&#x304A;&#x3051;&#x308B; 19q13.2 CIC &#x907A;&#x4F1D;&#x5B50;&#x3092;&#x767A;&#x898B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x5909;&#x7570;&#x306E;&#x4ED6;&#x306E; 7 &#x306E; CIC &#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x3001;&#x3053;&#x3046;&#x3057;&#x3066;&#x660E;&#x3089;&#x304B;&#x306B; 13 &#x306E;&#x8FFD;&#x52A0;&#x816B;&#x760D;&#x306E;&#x30C7;&#x30A3;&#x30FC;&#x30D7; &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x306B;&#x3088;&#x3063;&#x3066;&#x691C;&#x8A3C;&#x3055;&#x308C;&#x305F;&#x3053;&#x308C;&#x3089;&#x306E;&#x767A;&#x898B;&#x3092;&#x8A2D;&#x5B9A;&#x306F;&#x3001;20/29 &#xFF08;69 &#xFF05;&#xFF09; &#x672C;&#x7814;&#x7A76;&#x3067;&#x306F; oligodendroglioma &#x3067;&#x5168;&#x4F53;&#x7684;&#x306A;&#x7A81;&#x7136;&#x5909;&#x7570;&#x7387;&#x3092;&#x3082;&#x305F;&#x3089;&#x3059;&#x3053;&#x3068;&#x3002;&#x5BFE;&#x7167;&#x7684;&#x306B;&#x3001;CIC &#x306E;&#x5909;&#x5316;&#x304C;&#x5225;&#x3060;&#x3063;&#x305F;&#x3089;&#x816B;&#x3068; oligoastrocytomas 1 p/19q &#x640D;&#x5931;&#x306A;&#x3057;&#x306E;&#x30C7;&#x30A3;&#x30FC;&#x30D7; &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x73CD;&#x3057;&#x3044; (1/60; 2%)&#x3002;21 &#x975E;&#x540C;&#x7FA9;&#x4F53;&#x7D30;&#x80DE;&#x5909;&#x7570;&#x3092; 20 CIC &#x5909;&#x7570;&#x4F53; oligodendroglioma &#x306B;&#x304A;&#x3051;&#x308B;&#x30CA;&#x30A4;&#x30F3; &#x30A8;&#x30AF;&#x30BD;&#x30F3; 5 &#x6CE8;&#x91C8;&#x4ED8;&#x304D;&#x306E; DNA &#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x5185;&#x3067;&#x3001;3 &#x3064;&#x306E;&#x30A8;&#x30AF;&#x30BD;&#x30F3; 20 &#x6CE8;&#x91C8;&#x4ED8;&#x304D;&#x306E;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x306E;&#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x5185;&#x3067;&#x3060;&#x3063;&#x305F;&#x3002;&#x6B8B;&#x308A;&#x306E; 9 &#x4ED6;&#x306E;&#x30A8;&#x30AF;&#x30BD;&#x30F3;&#x3067;&#x767A;&#x898B;&#x3055;&#x308C;&#x305F;&#x3001;&#x983B;&#x7E41;&#x306B;&#x5207;&#x308A;&#x6368;&#x3066;&#x304C;&#x542B;&#x307E;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;CIC &#x5909;&#x7570;&#x3053;&#x3046;&#x3057;&#x3085;&#x7D44;&#x7E54;&#x5B66;, 1 p/19q co-deletion IDH1/2 &#x7A81;&#x7136;&#x5909;&#x7570; (p &lt; 0.001&#xFF09; &#x3068;&#x975E;&#x5E38;&#x306B;&#x95A2;&#x9023;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;&#x6211;&#x3005; &#x306E; CIC &#x5909;&#x7570;&#x4F53;&#x5BFE; 19q/1 p co-deletion &#x306E;&#x80CC;&#x666F;&#x306B;&#x306F;&#x3001;&#x91CE;&#x751F;&#x578B;&#x816B;&#x760D;&#x306E;&#x81E8;&#x5E8A;&#x7D50;&#x679C;&#x306E;&#x9055;&#x3044;&#x3092;&#x8A8D;&#x3081;&#x306A;&#x3044;&#x304C; hemizygous CIC &#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x306F;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308B;&#x3053;&#x3068;&#x304C;&#x91CD;&#x8981;&#x3002;&#x5358;&#x4E00;&#x4FDD;&#x6709; 19q &#x30A2;&#x30EC;&#x30EB;&#x5909;&#x7570; CIC oligodendroglioma IDH &#x5909;&#x7570;&#x306E;&#x75C5;&#x614B;&#x306B;&#x30EA;&#x30F3;&#x30AF;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x3092;&#x4EEE;&#x5B9A;&#x3057;&#x307E;&#x3059;&#x3002;&#x79C1;&#x305F;&#x3061;&#x3053;&#x3046;&#x3057;&#x3085;&#x907A;&#x4F1D;&#x7684;&#x7570;&#x5E38;&#x306E;&#x8A73;&#x7D30;&#x306A;&#x7814;&#x7A76;&#x3067;&#x306F; CIC &#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x3001;IDH1/2 &#x5909;&#x7570;&#x3068; 1 p/19q co-deletion &#x9593;&#x306E;&#x6A5F;&#x80FD;&#x7684;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3092;&#x793A;&#x5506;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#x540C;&#x6642; CIC &#x5909;&#x7570;&#x3068; IDH &#x5909;&#x7570;&#x3068; 1 p/19q &#x640D;&#x5931; oligodendroglioma &#x4ED6;&#x306E;&#x764C;&#x304B;&#x3089;&#x533A;&#x5225;&#x3057;&#x307E;&#x3059;&#x3002;

Oligodendroglioma &#xC784;&#xC0C1;, &#xBCD1; &#xC801;&#xC778;, &#xADF8;&#xB9AC;&#xACE0; &#xC720;&#xC804;&#xC801; &#xD2B9;&#xC9D5;&#xC5D0; &#xC758;&#xD574; &#xD2B9;&#xC9D5;&#xC785;&#xB2C8;&#xB2E4;. &#xC5FC;&#xC0C9;&#xCCB4; 1 p&#xC640; 19q&#xC758; &#xC7AC;&#xBC1C; &#xC131; &#xC190;&#xC2E4; &#xB1CC;&#xC885;&#xC591;&#xC774; &#xAC15;&#xB825; &#xD558; &#xAC8C; &#xC5F0;&#xACB0; &#xB418;&#xC5B4; &#xC788;&#xC9C0;&#xB9CC; id &#xC9C0;&#xC2DD;&#xACFC; &#xC774;&#xB7EC;&#xD55C; &#xBCC0;&#xACBD;&#xC5D0; &#xC758;&#xD574; &#xC601;&#xD5A5;&#xC744; &#xBC1B;&#xB294; &#xC720;&#xC804;&#xC790;&#xC758; &#xAE30;&#xB2A5; &#xC81C;&#xD55C; &#xB429;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; 1 p/19q co-deletion &#xC790;&#xB8CC; &#xC30D; &#xD574;&#xC0C1;&#xB3C4;&#xC5D0;&#xC11C; &#xC0C8;&#xB85C;&#xC6B4; &#xBD84;&#xC790; &#xAE30;&#xB2A5;&#xC744; &#xAC00;&#xC9C4; 16 oligodendrogliomas&#xC758; &#xAC80;&#xC0C9; &#xC9D1;&#xD569;&#xC5D0; exome &#xC2DC;&#xD000;&#xC2F1;&#xC744; &#xC218;&#xD589;. IDH &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;&#xC758; &#xB192;&#xC740; &#xC18D;&#xB3C4; &#xC608;&#xC0C1; &#xB300;&#xB85C;,: &#xBAA8;&#xB4E0; &#xACBD;&#xC6B0;&#xC5D0; &#xC5B4;&#xB290; IDH1 &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xD588;&#xB2E4; (14/16) &#xB610;&#xB294; IDH2 (2/16). &#xB610;&#xD55C;, &#xC6B0;&#xB9AC; 13/16 &#xC885;&#xC591;&#xC5D0;&#xC11C; &#xC5FC;&#xC0C9;&#xCCB4; 19q13.2&#xC5D0; CIC &#xC720;&#xC804;&#xC790;&#xC5D0; &#xC0BD;&#xC785;/&#xC0AD;&#xC81C; &#xCCB4;&#xC138;&#xD3EC; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xBC1C;&#xACAC;. &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; CIC &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;&#xC640; 7 &#xAC1C;&#xC758; &#xB2E4;&#xB978; &#xC0AC;&#xB78C;&#xB4E4;&#xC744; &#xB530;&#xB77C;&#xC11C; &#xACF5;&#xAC1C; 13 &#xCD94;&#xAC00; &#xC885;&#xC591;&#xC758; &#xAE4A;&#xC740; &#xC5F0;&#xC18D; &#xD655;&#xC778; &#xD588;&#xB2E4; &#xC774;&#xB7EC;&#xD55C; &#xB514;&#xC2A4;&#xCEE4;&#xBC84;&#xB9AC; &#xC138;&#xD2B8; 20/29 (69%)&#xC774;&#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; oligodendrogliomas &#xC804;&#xCCB4; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC728;&#xC744; &#xB370;&#xB9AC;&#xACE0;. &#xB300;&#xC870;&#xC801;&#xC73C;&#xB85C;, astrocytomas &#xBC0F; oligoastrocytomas 1 p/19q &#xC190;&#xC2E4; &#xC5C6;&#xC774; &#xAE4A;&#xC740; &#xC2DC;&#xD000;&#xC2F1; CIC &#xBCC0;&#xACBD; &#xD588;&#xB2E4; &#xADF8;&#xB807;&#xC9C0; &#xC54A;&#xC73C;&#xBA74; &#xACF5;&#xAC1C; &#xD76C;&#xADC0; (1/60, 2%). 20 CIC &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; oligodendrogliomas&#xC5D0;&#xC11C; 21-&#xB300;&#xBA85;&#xC0AC; &#xCCB4;&#xC138;&#xD3EC; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;, 9 exon 5 &#xC8FC;&#xC11D;&#xC774; &#xB2EC;&#xB9B0;&#xB41C; DNA &#xC0C1;&#xD638; &#xC791;&#xC6A9; &#xB3C4;&#xBA54;&#xC778; &#xB0B4;&#xC5D0; &#xC788;&#xC5C8;&#xB294;&#xB370; 3 exon 20 &#xC8FC;&#xC11D;&#xB41C; &#xB2E8;&#xBC31;&#xC9C8; &#xC0C1;&#xD638; &#xC791;&#xC6A9; &#xB3C4;&#xBA54;&#xC778; &#xB0B4;&#xC5D0; &#xD588;&#xB2E4;. &#xB098;&#xBA38;&#xC9C0; 9 &#xB2E4;&#xB978; exons&#xC5D0;&#xC11C; &#xBC1C;&#xACAC; &#xD558; &#xACE0; &#xC790;&#xC8FC; &#xC798;&#xB9BC; &#xD3EC;&#xD568;. CIC &#xBCC0;&#xC774; &#xB9E4;&#xC6B0; oligodendroglioma &#xC870;&#xC9C1;&#xD559;, 1 p/19q co-deletion IDH1/2 &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; (p &lt; 0.001)&#xC640; &#xAD00;&#xB828; &#xB41C; &#xD588;&#xB2E4;. Hemizygous CIC &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xD655;&#xB960;&#xC774; &#xBE44;&#xB85D; &#xC6B0;&#xB9AC;&#xAC00; &#xAD00;&#xCC30; 1 p/19q co-deletion &#xBC30;&#xACBD;&#xC5D0;&#xC11C; &#xC57C;&#xC0DD; &#xD615; &#xC885;&#xC591; &#xB300; CIC &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;&#xC758; &#xC784;&#xC0C1; &#xACB0;&#xACFC;&#xC5D0; &#xCC28;&#xC774;&#xAC00; &#xC911;&#xC694; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xB2E8;&#xC77C; &#xBCF4;&#xAD00;&#xB41C; 19q &#xB300;&#xB9BD; &#xC720;&#xC804;&#xC790;&#xC5D0; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; CIC IDH &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;&#xC640; oligodendrogliomas&#xC758; pathogenesis &#xC5F0;&#xACB0; &#xAC00;&#xC124;. &#xC6B0;&#xB9AC;&#xC758; &#xC0C1;&#xC138;&#xD55C; &#xC5F0;&#xAD6C; oligodendroglioma &#xC720;&#xC804;&#xC790; &#xCC29;&#xC624;&#xC758; &#xAE30;&#xB2A5;&#xC801; &#xC0C1;&#xD638;&#xC791;&#xC6A9;&#xC744; CIC &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;, IDH1/2 &#xBCC0;&#xC774; 1 p/19q co-deletion &#xB098;&#xC654;&#xB2E4;.

&#xB3D9;&#xC2DC; CIC &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;, IDH &#xBCC0;&#xC774; 1 p/19q &#xC190;&#xC2E4; &#xB2E4;&#xB978; &#xC554;&#xC5D0;&#xC11C; oligodendrogliomas&#xB97C; &#xAD6C;&#xBCC4; &#xD569;&#xB2C8;&#xB2E4;.

Oligodendroglioma se caracteriza por las caracter&iacute;sticas cl&iacute;nicas, patol&oacute;gicas y gen&eacute;ticas &uacute;nicas. P&eacute;rdidas recurrentes de los cromosomas 1P y 19q est&aacute;n fuertemente asociadas con este c&aacute;ncer en el cerebro pero el conocimiento de la identidad y funci&oacute;n de los genes afectados por estas alteraciones se limita. Realizamos la secuencia del exoma en un conjunto de descubrimiento de 16 oligodendrogliomas con 1 p/19q codeleci&oacute;n para identificar nuevas caracter&iacute;sticas moleculares en resoluci&oacute;n de base-par. Seg&uacute;n lo previsto, hubo una alta tasa de mutaciones de IDH: todos los casos ten&iacute;an mutaciones en ambos IDH1 (14/16) o IDH2 (2/16). Adem&aacute;s, hemos descubierto las mutaciones som&aacute;ticas e inserciones/deleciones en el gen de la CIC en el cromosoma 19q13.2 en tumores de 13/16. Esta serie de descubrimiento mutaciones fueron validadas por la secuencia profunda de 13 tumores adicionales, que revel&oacute; otros siete con mutaciones de la CIC, as&iacute; que la tasa de mutaci&oacute;n en oligodendrogliomas en este estudio 20/29 (69%). Por el contrario, la secuencia profunda de astrocitomas y oligoastrocytomas sin p&eacute;rdida de p/19q 1 revel&oacute; que alteraciones de la CIC que de otro modo rara (1/60; 2%). De las 21 mutaciones som&aacute;ticas no sin&oacute;nimas en 20 CIC-mutante oligodendrogliomas, nueve eran en el ex&oacute;n 5 dentro de un dominio de interacci&oacute;n DNA anotado y tres fueron en el ex&oacute;n 20 dentro de un dominio de interacci&oacute;n con la prote&iacute;na anotado. Los nueve restantes fueron encontrados en otros exones y con frecuencia incluyen truncamientos. CIC mutaciones eran altamente asociadas con histolog&iacute;a de oligodendroglioma, 1 codeleci&oacute;n p/19q y mutaci&oacute;n IDH1/2 (p &lt; 0.001). Aunque no se observaron diferencias en los resultados cl&iacute;nicos del mutante CIC frente a tumores de tipo salvaje, en un fondo de codeleci&oacute;n 1 p/19q, hemizygous CIC mutaciones son probablemente importantes. Presumimos que el mutante CIC en el alelo 19q retenidas solo est&aacute; ligado a la patogenesia de oligodendrogliomas con mutaci&oacute;n del IDH. Nuestro estudio detallado de las aberraciones gen&eacute;ticas en oligodendroglioma sugiere una interacci&oacute;n funcional entre la mutaci&oacute;n de la CIC, mutaci&oacute;n IDH1/2 y 1 codeleci&oacute;n p/19q.

Mutaciones de CIC concurrentes, mutaciones de IDH y 1 p/19q p&eacute;rdida distinguen oligodendrogliomas de otros tipos de c&aacute;ncer.

Somatic mutations in the catalytic domain of isocitrate dehydrogenase (IDH) 1/2 and accumulation of the oncometabolite 2-hydroxyglutarate (2-HG) appear to be among the earliest events in gliomagenesis and may contribute to malignant transformation. The lack of cell lines with endogenous mutations has been one of the major challenges in studying IDH1/2-mutant glioma and developing novel therapeutics for these tumors. Here, we describe the isolation of a glioma brain tumor stem cell line (BT142) with an endogenous R132H mutation in IDH1, aggressive tumor-initiating capacity, and 2-HG production. The neurosphere culture method was used to establish a brain tumor stem cell line from an IDH1-mutant anaplastic oligoastrocytoma sample, and an orthotopic xenograft system was developed to allow its rapid expansion. Production of 2-HG by glioma cells with endogenous IDH1 mutations was confirmed by mass spectrometry. BT142 retained an endogenous R132H IDH1 mutation in culture and possessed aggressive tumor-initiating capacity, allowing it to be readily propagated in orthotopic xenografts of nonobese diabetic/severe combined immune deficiency (NOD SCID) mice. Endogenous 2-HG production by BT142 was detectable in both cell culture medium and xenograft animal serum. BT142 is the first brain tumor cell line with an endogenous IDH1 mutation and detectable 2-HG production both in vitro and in vivo, which thus provides a unique model for studying the biology of IDH1-mutant glioma and in vivo validation of compounds targeting IDH1-mutant cells.

An in vivo patient-derived model of endogenous IDH1-mutant glioma.

Oligodendroglioma is characterized by mutations of IDH and CIC, 1p/19q loss, and slow growth. We found that NHE-1 on 1p is silenced in oligodendrogliomas secondary to IDH-associated hypermethylation and 1p allelic loss. Silencing lowers intracellular pH and attenuates acid load recovery in oligodendroglioma cells. Others have shown that rapid tumor growth cannot occur without NHE-1-mediated neutralization of the acidosis generated by the Warburg glycolytic shift. Our findings show for the first time that the pH regulator NHE-1 can be silenced in a human cancer and also suggest that pH deregulation may contribute to the distinctive biology of human oligodendroglioma.

DNA hypermethylation and 1p Loss silence NHE-1 in oligodendroglioma.

Spontaneous loss of heterozygosity leading to homozygous R132H in a patient-derived IDH1 mutant cell line.

In glioblastoma multiforme (GBM), brain-tumor-initiating cells (BTICs) with cancer stem cell characteristics have been identified and proposed as primordial cells responsible for disease initiation, recurrence, and therapeutic resistance. However, the extent to which individual, patient-derived BTIC lines reflect the heterogeneity of GBM remains poorly understood. Here we applied a stem cell biology approach and compared self-renewal, marker expression, label retention, and asymmetric cell division in 20 BTIC lines. Through cluster analysis, we identified two subgroups of BTIC lines with distinct precursor states, stem- or progenitor-like, predictive of survival after xenograft. Moreover, stem and progenitor transcriptomic signatures were identified, which showed a strong association with the proneural and mesenchymal subtypes, respectively, in the TCGA cohort. This study proposes a different framework for the study and use of BTIC lines and provides precursor biology insights into GBM. 

Precursor States of Brain Tumor Initiating Cell Lines Are Predictive of Survival in Xenografts and Associated with Glioblastoma Subtypes.

 mutation is the earliest genetic alteration in low-grade gliomas (LGGs), but its role in tumor recurrence is unclear. Mutant IDH1 drives overproduction of the oncometabolite d-2-hydroxyglutarate (2HG) and a CpG island (CGI) hypermethylation phenotype (G-CIMP). To investigate the role of mutant IDH1 at recurrence, we performed a longitudinal analysis of 50  mutant LGGs. We discovered six cases with copy number alterations (CNAs) at the  locus at recurrence. Deletion or amplification of  was followed by clonal expansion and recurrence at a higher grade. Successful cultures derived from  mutant, but not  wild type, gliomas systematically deleted  in vitro and in vivo, further suggestive of selection against the heterozygous mutant state as tumors progress. Tumors and cultures with  CNA had decreased 2HG, maintenance of G-CIMP, and DNA methylation reprogramming outside CGI. Thus, while  mutation initiates gliomagenesis, in some patients mutant IDH1 and 2HG are not required for later clonal expansions.

Article	Year
Concurrent CIC mutations, IDH mutations, and 1p/19q loss distinguish oligodendrogliomas from other cancers. The Journal of pathology\| PubMed ID: 22072542	2012
An in vivo patient-derived model of endogenous IDH1-mutant glioma. Neuro-oncology\| PubMed ID: 22166263	2012
DNA hypermethylation and 1p Loss silence NHE-1 in oligodendroglioma. Annals of neurology\| PubMed ID: 22718548	2012
Spontaneous loss of heterozygosity leading to homozygous R132H in a patient-derived IDH1 mutant cell line. Neuro-oncology\| PubMed ID: 23757293	2013
Precursor States of Brain Tumor Initiating Cell Lines Are Predictive of Survival in Xenografts and Associated with Glioblastoma Subtypes. Stem cell reports\| PubMed ID: 26095605	2015
Clonal expansion and epigenetic reprogramming following deletion or amplification of mutant . Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 28916733	2017

Charles Chesnelong

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