Isolation of Distinct Cell Populations from the Developing Cerebellum by Microdissection

Microdissection is a novel technique that can isolate specific regions of a tissue and eliminate contamination from cellular sources in adjacent areas. ...

Temple University Health System

The advent of techniques for global analyses of cell biology, such as genomics and proteomics, opens the way to rapid progress in understanding the molecular control of developing tissues. However, such studies in the CNS are hindered by the complexity of this tissue. In particular, few approaches allow cells to be isolated that are enriched for specific stages of their maturation. We describe a new strategy to study gene expression and function in cerebellar granule cells. In these experiments, we have used square pulse electroporation to introduce fluorescent dye or DNA constructs into immature granule cell precursors in situ. This method only labels granule cell precursors in the superficial part of the external granule layer. Combining this labelling with fluorescent activated cell sorting (FACS) allows the transfected cells to be isolated at any time during their subsequent development, thus providing a means of analysing granule cells as they undergo maturation. This transfection method can be used to study events in the normal maturation of granule cells or the effects of introduced transgenes. Such studies can be carried out on cells purified from primary cultures or cells in situ using cerebellar slice cultures. Our strategy provides a new route to detailed analysis of the role of genes in controlling many aspects of granule cell biology. These approaches will allow recent global analyses to be more readily applied to subpopulations of cells in complex tissues.

Novel strategy to study gene expression and function in developing cerebellar granule cells.

Cancer stem cells (CSCs) are thought to be critical for initiation and propagation of many types of cancer. Because these cells are resistant to conventional therapies, they have been very difficult to eliminate. A study in this issue of Cancer Cell suggests that brain tumor CSCs live in a "vascular niche" that promotes their long-term growth and self-renewal. Disrupting this niche impairs CSC self-renewal and thereby significantly inhibits the growth of tumors. Targeting the unique microenvironment of CSCs may be the key to effective cancer therapy.

Hit 'em where they live: targeting the cancer stem cell niche.

To provide an ideal method for flap prefabrication.

[Repair of the facial defects using the expanded induced prefabricated skin flap of the retroauricular and mastoid process region based on the superficial temporal vascular bundle].

To apply the scapular free flap extended to the upper arm for resurfacing the face and neck, as well as the upper lip in one stage.

[Clinical application of the scapular free flap extended to the upper arm].

Medulloblastoma is the most common malignant brain tumor in children, but the cells from which it arises remain unclear. Here we examine the origin of medulloblastoma resulting from mutations in the Sonic hedgehog (Shh) pathway. We show that activation of Shh signaling in neuronal progenitors causes medulloblastoma by 3 months of age. Shh pathway activation in stem cells promotes stem cell proliferation but only causes tumors after commitment to-and expansion of-the neuronal lineage. Notably, tumors initiated in stem cells develop more rapidly than those initiated in progenitors, with all animals succumbing by 3-4 weeks. These studies suggest that medulloblastoma can be initiated in progenitors or stem cells but that Shh-induced tumorigenesis is associated with neuronal lineage commitment.

Medulloblastoma can be initiated by deletion of Patched in lineage-restricted progenitors or stem cells.

Studying the early stages of cancer can provide important insight into the molecular basis of the disease. We identified a preneoplastic stage in the patched (ptc) mutant mouse, a model for the brain tumor medulloblastoma. Preneoplastic cells (PNCs) are found in most ptc mutants during early adulthood, but only 15% of these animals develop tumors. Although PNCs are found in mice that develop tumors, the ability of PNCs to give rise to tumors has never been demonstrated directly, and the fate of cells that do not form tumors remains unknown. Using genetic fate mapping and orthotopic transplantation, we provide definitive evidence that PNCs give rise to tumors, and show that the predominant fate of PNCs that do not form tumors is differentiation. Moreover, we show that N-myc, a gene commonly amplified in medulloblastoma, can dramatically alter the fate of PNCs, preventing differentiation and driving progression to tumors. Importantly, N-myc allows PNCs to grow independently of hedgehog signaling, making the resulting tumors resistant to hedgehog antagonists. These studies provide the first direct evidence that PNCs can give rise to tumors, and demonstrate that identification of genetic changes that promote tumor progression is critical for designing effective therapies for cancer.

N-myc alters the fate of preneoplastic cells in a mouse model of medulloblastoma.

Medulloblastoma encompasses a collection of clinically and molecularly diverse tumour subtypes that together comprise the most common malignant childhood brain tumour. These tumours are thought to arise within the cerebellum, with approximately 25% originating from granule neuron precursor cells (GNPCs) after aberrant activation of the Sonic Hedgehog pathway (hereafter, SHH subtype). The pathological processes that drive heterogeneity among the other medulloblastoma subtypes are not known, hindering the development of much needed new therapies. Here we provide evidence that a discrete subtype of medulloblastoma that contains activating mutations in the WNT pathway effector CTNNB1 (hereafter, WNT subtype) arises outside the cerebellum from cells of the dorsal brainstem. We found that genes marking human WNT-subtype medulloblastomas are more frequently expressed in the lower rhombic lip (LRL) and embryonic dorsal brainstem than in the upper rhombic lip (URL) and developing cerebellum. Magnetic resonance imaging (MRI) and intra-operative reports showed that human WNT-subtype tumours infiltrate the dorsal brainstem, whereas SHH-subtype tumours are located within the cerebellar hemispheres. Activating mutations in Ctnnb1 had little impact on progenitor cell populations in the cerebellum, but caused the abnormal accumulation of cells on the embryonic dorsal brainstem which included aberrantly proliferating Zic1(+) precursor cells. These lesions persisted in all mutant adult mice; moreover, in 15% of cases in which Tp53 was concurrently deleted, they progressed to form medulloblastomas that recapitulated the anatomy and gene expression profiles of human WNT-subtype medulloblastoma. We provide the first evidence, to our knowledge, that subtypes of medulloblastoma have distinct cellular origins. Our data provide an explanation for the marked molecular and clinical differences between SHH- and WNT-subtype medulloblastomas and have profound implications for future research and treatment of this important childhood cancer.

Subtypes of medulloblastoma have distinct developmental origins.

&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x5305;&#x62EC;&#x5171;&#x540C;&#x6784;&#x6210;&#x6700;&#x5E38;&#x89C1;&#x7684;&#x513F;&#x7AE5;&#x6076;&#x6027;&#x8111;&#x80BF;&#x7624;&#x7684;&#x4E34;&#x5E8A;&#x548C;&#x5206;&#x5B50;&#x4E0D;&#x540C;&#x80BF;&#x7624;&#x5B50;&#x7C7B;&#x578B;&#x7684;&#x96C6;&#x5408;&#x3002;&#x636E;&#x8BA4;&#x4E3A;&#x8FD9;&#x4E9B;&#x80BF;&#x7624;&#x5185;&#x5C0F;&#x8111;&#xFF0C;&#x7EA6; 25%&#x6765;&#x81EA;&#x9897;&#x7C92;&#x795E;&#x7ECF;&#x5143;&#x524D;&#x4F53;&#x7EC6;&#x80DE; (GNPCs) &#x4E4B;&#x540E;&#xFF0C;&#x4F1A;&#x51FA;&#x73B0;&#x5F02;&#x5E38;&#x6D3B;&#x5316;&#x7684;&#x58F0;&#x6CE2;&#x523A;&#x732C;&#x901A;&#x8DEF; &#xFF08;&#x4EE5;&#x4E0B;&#x7B80;&#x79F0; SHH &#x5B50;&#x7C7B;&#x578B;&#xFF09;&#x3002;&#x9A71;&#x52A8;&#x5668;&#x95F4;&#x7684;&#x5B50;&#x7C7B;&#x578B;&#x672A;&#x77E5;&#x7684;&#x5176;&#x4ED6;&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x7684;&#x5F02;&#x8D28;&#x6027;&#x7684;&#x75C5;&#x7406;&#x8FC7;&#x7A0B;&#xFF0C;&#x963B;&#x788D;&#x53D1;&#x5C55;&#x7684;&#x5F88;&#x591A;&#x9700;&#x8981;&#x65B0;&#x7684;&#x7597;&#x6CD5;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x8BC1;&#x636E;&#x7684;&#x80CC;&#x4FA7;&#x8111;&#x5E72;&#x7EC6;&#x80DE;&#x5916;&#x5C0F;&#x8111;&#x5F15;&#x8D77;&#x7684;&#x79BB;&#x6563;&#x5B50;&#x7C7B;&#x578B;&#x5305;&#x542B;&#x6D3B;&#x5316;&#x7A81;&#x53D8; WNT &#x901A;&#x8DEF;&#x6548;&#x5E94;&#x5668; CTNNB1 &#xFF08;&#x4EE5;&#x4E0B;&#x7B80;&#x79F0; WNT &#x5B50;&#x7C7B;&#x578B;&#xFF09; &#x7684;&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x3002;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x66F4;&#x9891;&#x7E41;&#x8868;&#x793A;&#x6807;&#x8BB0;&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE; WNT &#x5B50;&#x7C7B;&#x578B;&#x4EBA;&#x7C7B;&#x7684;&#x57FA;&#x56E0;&#x662F;&#x83F1;&#x5F62;&#x4E0B;&#x5507; (LRL) &#x548C;&#x80DA;&#x80CE;&#x80CC;&#x8111;&#x5E72;&#x4E2D;&#x6BD4;&#x5728;&#x53F3;&#x4E0A;&#x89D2;&#x83F1;&#x5F62;&#x5507; (URL) &#x548C;&#x53D1;&#x5C55;&#x4E2D;&#x56FD;&#x5BB6;&#x5C0F;&#x8111;&#x3002;&#x78C1;&#x5171;&#x632F;&#x6210;&#x50CF; (MRI) &#x548C;&#x672F;&#x4E2D;&#x62A5;&#x544A;&#x8868;&#x660E;&#x4EBA;&#x7C7B; WNT &#x5B50;&#x7C7B;&#x578B;&#x80BF;&#x7624;&#x6E17;&#x5165;&#x80CC;&#x8111;&#x5E72;&#xFF0C;&#x800C; SHH &#x5B50;&#x7C7B;&#x578B;&#x80BF;&#x7624;&#x4F4D;&#x4E8E;&#x5C0F;&#x8111;&#x534A;&#x7403;&#x5185;&#x3002;&#x6FC0;&#x6D3B; Ctnnb1 &#x7A81;&#x53D8;&#x5C0F;&#x8111;&#x7684;&#x7956;&#x7EC6;&#x80DE;&#x7FA4;&#x4F53;&#x8FC7;&#x5927;&#x5F71;&#x54CD;&#xFF0C;&#x4F46;&#x5BF9;&#x5176;&#x4E2D;&#x5305;&#x62EC;&#x95ED;&#x5408;&#x589E;&#x6B96; Zic1(+) &#x4F53;&#x7EC6;&#x80DE;&#x80DA;&#x80CE;&#x80CC;&#x8111;&#x5E72;&#x9020;&#x6210;&#x7EC6;&#x80DE;&#x7684;&#x5F02;&#x5E38;&#x79EF;&#x805A;&#x3002;&#x8FD9;&#x4E9B;&#x75C5;&#x53D8;&#x4FDD;&#x7559;&#x6240;&#x6709;&#x7A81;&#x53D8;&#x4F53;&#x7684;&#x6210;&#x5E74;&#x5C0F;&#x9F20; &#xFF1B;&#x6B64;&#x5916;&#xFF0C;15%&#x7684;&#x60C5;&#x51B5;&#x4E0B;&#x540C;&#x65F6;&#x5220;&#x9664; Tp53 &#x4E86;&#xFF0C;&#x4ED6;&#x4EEC;&#x8FDB;&#x5C55;&#x5230;&#x6982;&#x62EC; WNT &#x5B50;&#x7C7B;&#x578B;&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x89E3;&#x5256;&#x548C;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x8C31;&#x7684;&#x5F62;&#x5F0F;&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE;&#x3002;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x7684;&#x7B2C;&#x4E00;&#x4E2A;&#x8BC1;&#x636E;&#xFF0C;&#x5C31;&#x6211;&#x4EEC;&#x6240;&#x77E5;&#xFF0C;&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x7684;&#x4E9A;&#x578B;&#x6709;&#x4E0D;&#x540C;&#x7684;&#x7EC6;&#x80DE;&#x6765;&#x6E90;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x6570;&#x636E;&#x63D0;&#x4F9B;&#x6807;&#x8BB0;&#x5206;&#x5B50;&#x4E0E;&#x4E34;&#x5E8A;&#x4E4B;&#x95F4;&#x7684;&#x5DEE;&#x5F02; SHH &#x5B50;&#x548C; WNT &#x7C7B;&#x578B;&#x9AD3;&#x6BCD;&#x89E3;&#x91CA;&#x548C;&#x5BF9;&#x672A;&#x6765;&#x7684;&#x7814;&#x7A76;&#x548C;&#x6CBB;&#x7597;&#x6B64;&#x91CD;&#x8981;&#x7AE5;&#x5E74;&#x764C;&#x75C7;&#x5177;&#x6709;&#x6DF1;&#x8FDC;&#x5F71;&#x54CD;&#x3002;

&#x9AD3;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x7684;&#x4E9A;&#x578B;&#x6709;&#x4E0D;&#x540C;&#x7684;&#x53D1;&#x5C55;&#x6E0A;&#x6E90;&#x3002;

M&eacute;dulloblastome comprend une collection de sous-types de tumeurs diverses sur le plan clinique et mol&eacute;culaire qui ensemble constituent la tumeur de cerveau malignes de l&#39;enfance plus courante. On pense que ces tumeurs surviennent dans le cervelet, avec environ 25 % provenant de cellules pr&eacute;curseurs de granule neurone (GNPCs) apr&egrave;s l&#39;activation aberrante de la voie de Sonic Hedgehog (ci-apr&egrave;s, le sous-type SHH). Processus pathologiques qui gouvernent l&#39;h&eacute;t&eacute;rog&eacute;n&eacute;it&eacute; parmi l&#39;autre m&eacute;dulloblastome sous-types ne sont pas connus, emp&ecirc;chent le d&eacute;veloppement d&#39;une grande partie avait besoin de nouvelles th&eacute;rapies. Nous fournissons ici la preuve qu&#39;un sous-type discr&egrave;te d&#39;un m&eacute;dulloblastome qui contient des mutations activante l&#39;effecteur de voie WNT CTNNB1 (ci-apr&egrave;s, le sous-type WNT) se pose en dehors du cervelet des cellules de tronc c&eacute;r&eacute;bral dorsal. Nous avons trouv&eacute; que g&egrave;nes marquage humaine WNT-sous-type m&eacute;dulloblastomes sont plus fr&eacute;quemment exprim&eacute;s dans la l&egrave;vre sup&eacute;rieure rhombique (LRL) et le tronc dorsal embryonnaire que dans la partie sup&eacute;rieure rhombique l&egrave;vre (URL) et le d&eacute;veloppement de cervelet. L&#39;imagerie par r&eacute;sonance magn&eacute;tique (IRM) et les rapports perop&eacute;ratoires a montr&eacute; que des tumeurs humaines de WNT-sous-type s&#39;infiltrer dans le tronc c&eacute;r&eacute;bral dorsal, tandis que les tumeurs de SHH-sous-type sont situ&eacute;s dans les h&eacute;misph&egrave;res c&eacute;r&eacute;belleuses. Activation des mutations Ctnnb1 eu peu d&#39;impact sur les populations de cellules prog&eacute;nitrices dans le cervelet, mais caus&eacute; l&#39;accumulation anormale de cellules sur le tronc c&eacute;r&eacute;bral dorsal embryonnaire qui comprenait des cellules de pr&eacute;curseur aberrantly prolif&eacute;ration Zic1(+). Ces l&eacute;sions ont persist&eacute; dans toutes les souris adultes mutantes ; en outre, dans 15 % des cas dans lesquels Tp53 &eacute;tait en m&ecirc;me temps supprim&eacute;, l&#39;on avancerait &agrave; forme m&eacute;dulloblastomes qui r&eacute;capitule les profils d&#39;expression anatomie et g&egrave;ne de m&eacute;dulloblastome de WNT-sous-type humain. Nous fournissent la premi&egrave;re preuve, &agrave; notre connaissance, que les sous-types de m&eacute;dulloblastome ont des origines cellulaires distincts. Nos r&eacute;sultats fournissent une explication pour les cliniques et mol&eacute;culaires des diff&eacute;rences marqu&eacute;es entre les m&eacute;dulloblastomes SHH et WNT-sous-type et ont des implications profondes pour les futures recherches et le traitement de ce cancer infantile important.

Sous-types de m&eacute;dulloblastome ont des origines distinctes de d&eacute;veloppement.

Medulloblastom umfasst eine Sammlung von klinisch und molekular unterschiedlichen Tumor-Subtypen, die zusammen die h&auml;ufigste b&ouml;sartige Kindheit-Hirntumor bilden. Diese Tumoren werden gedacht, um innerhalb des Kleinhirns mit ca. 25 % aus Granulat Neuron Vorl&auml;uferzellen (GNPCs) nach aberrante Aktivierung des Sonic Hedgehog-Signalweges (nachstehend, SHH Subtype) auftreten. Die pathologischen Prozesse, die Heterogenit&auml;t unter den anderen Medulloblastom fahren, Subtypen nicht bekannt sind, behindern die Entwicklung der viel ben&ouml;tigt neue Therapien. Hier stellen wir Hinweise, dass ein diskreter Untertyp von Medulloblastom, die aktivierende Mutationen in den WNT-Signalweg-Effektor CTNNB1 (nachstehend, WNT Subtype) enth&auml;lt au&szlig;erhalb des Kleinhirns aus Zellen des dorsalen Hirnstamms entsteht. Wir fanden, dass Gene, die Kennzeichnung der menschlichen WNT-Subtyp Medulloblastomas h&auml;ufiger in der rhombischen Unterlippe (LRL) und embryonalen dorsalen Hirnstamm ausgedr&uuml;ckt werden, als in der oberen rhombische Lippe (URL) und die Entwicklung des Kleinhirns. Magnetresonanz-Bildgebung (MRI) und intraoperative Berichte zeigten, dass menschliche WNT-Untertyp Tumoren dorsalen Hirnstamm infiltrieren, w&auml;hrend SHH-Untertyp Tumore innerhalb der Kleinhirnhemisph&auml;ren befinden. Aktivierung von Mutationen im Ctnnb1 hatte eine geringe Auswirkung auf die Vorfahr Zelle Bev&ouml;lkerung im Kleinhirn, aber verursacht die abnorme Ansammlung von Zellen auf der embryonalen dorsalen Hirnstamm das abnorm wuchernden Zic1(+) Vorl&auml;uferzellen enthalten. Diese L&auml;sionen beibehalten in allen mutierten Erwachsenen M&auml;usen; Dar&uuml;ber hinaus fortgeschritten in 15 % der F&auml;lle, in denen Tp53 gleichzeitig gel&ouml;scht wurde, sie zu Form-Medulloblastomas, die die Anatomie und Gen-Ausdruck-Profile der menschlichen WNT-Untertyp Medulloblastom rekapituliert. Wir bieten dem ersten Beweis unseres Wissens Untertypen von Medulloblastom unterschiedliche zellul&auml;re Ursprung haben. Unsere Daten bieten eine Erkl&auml;rung f&uuml;r die deutliche Molekulare und klinische Unterschiede zwischen SHH und WNT-Subtyp Medulloblastomas und tiefgreifende Auswirkungen auf zuk&uuml;nftige Forschung und Behandlung von dieser wichtigen Kindheit-Krebs.

Untertypen von Medulloblastom haben verschiedene Entwicklungs Ursprung.

Medulloblastoma comprende un insieme dei sottotipi di tumore clinicamente e molecolarmente diverse che insieme costituiscono il pi&ugrave; comune tumore cerebrale infantile maligno. Questi tumori sono pensa che sorgono all&#39;interno del cervelletto, con circa il 25% provenienti da cellule precursori di granulo neurone (GNPCs) dopo l&#39;attivazione aberrante della via di Sonic Hedgehog (in appresso, SHH sottotipo). I processi patologici che guidano eterogeneit&agrave; tra il medulloblastoma altri sottotipi non sono noti, ostacolando lo sviluppo di gran parte necessarie nuove terapie. Qui forniamo prove che un sottotipo discreto di medulloblastoma che contiene mutazioni attivanti nel effector pathway WNT CTNNB1 (in appresso, sottotipo WNT) si pone di fuori del cervelletto dalle cellule del tronco encefalico dorsale. Abbiamo trovato che geni umani medulloblastomi WNT-sottotipo di marcatura sono pi&ugrave; frequentemente espresso nel labbro inferiore rombico (LRL) ed embrionale dorsale del tronco encefalico che nella parte superiore rombico del labbro (URL) e lo sviluppo di cervelletto. Risonanza magnetica (MRI) e le relazioni intra-operatoria mostrarono che tumori WNT-sottotipo umani infiltrano il dorsale del tronco encefalico, mentre tumori SHH-sottotipo si trovano gli emisferi cerebellari. Attivando le mutazioni Ctnnb1 ebbe poco impatto sulle popolazioni di cellule progenitrici nel cervelletto, ma ha causato l&#39;accumulo di cellule anomala sul tronco encefalico dorsale embrionale che comprendeva le cellule precursore in modo anomalo proliferazione Zic1(+). Queste lesioni permanenti in tutti i topi mutanti adulti; Inoltre, nel 15% dei casi in cui Tp53 &egrave; stato eliminato contemporaneamente, hanno progredito a forma medulloblastomi che ricapitolate profili di espressione di gene e anatomia di medulloblastoma umano di WNT-sottotipo. Forniamo la prima prova, a nostra conoscenza, che sottotipi di medulloblastoma hanno origini cellulari distinti. I nostri dati forniscono una spiegazione per le marcate differenze molecolari e cliniche tra SHH e WNT sottotipo medulloblastomi e hanno profonde implicazioni per la ricerca futura e il trattamento di questo tumore infantile importante.

Sottotipi di medulloblastoma hanno origini dello sviluppo distinti.

&#x9AC4;&#x82BD;&#x816B;&#x306B;&#x306F;&#x4E00;&#x7DD2;&#x306B;&#x6700;&#x3082;&#x4E00;&#x822C;&#x7684;&#x306A;&#x60AA;&#x6027;&#x5C0F;&#x5150;&#x8133;&#x816B;&#x760D;&#x3092;&#x69CB;&#x6210;&#x3059;&#x308B;&#x81E8;&#x5E8A;&#x304A;&#x3088;&#x3073;&#x5206;&#x5B50;&#x591A;&#x69D8;&#x306A;&#x816B;&#x760D;&#x306E;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x306E;&#x30B3;&#x30EC;&#x30AF;&#x30B7;&#x30E7;&#x30F3;&#x304C;&#x542B;&#x307E;&#x308C;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x816B;&#x760D;&#x306F;&#x5C0F;&#x8133;&#x5185;&#x304A;&#x3088;&#x305D; 25% &#x9846;&#x7C92;&#x30CB;&#x30E5;&#x30FC;&#x30ED;&#x30F3;&#x524D;&#x99C6;&#x7D30;&#x80DE; (GNPCs) &#x304B;&#x3089;&#x306E;&#x767A;&#x4FE1;&#x30BD;&#x30CB;&#x30C3;&#x30AF; &#x30D8;&#x30C3;&#x30B8;&#x30DB;&#x30C3;&#x30B0;&#x7D4C;&#x8DEF; (&#x4EE5;&#x4E0B;&#x3001;SHH &#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;) &#x306E;&#x7570;&#x5E38;&#x6D3B;&#x6027;&#x5316;&#x5F8C;&#x306B;&#x751F;&#x3058;&#x308B;&#x3068;&#x8003;&#x3048;&#x3089;&#x308C;&#x307E;&#x3059;&#x3002;&#x4E0D;&#x5747;&#x4E00;&#x6027;&#x306E;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x306F;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x306A;&#x3044;&#x4ED6;&#x306E;&#x9AC4;&#x9593;&#x3092;&#x904B;&#x8EE2;&#x3001;&#x75C5;&#x7406;&#x5B66;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x3001;&#x591A;&#x304F;&#x306E;&#x958B;&#x767A;&#x3092;&#x59A8;&#x3052;&#x308B;&#x65B0;&#x3057;&#x3044;&#x7642;&#x6CD5;&#x304C;&#x5FC5;&#x8981;&#x3002;&#x3053;&#x3053;&#x3067;&#x96E2;&#x6563;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7; WNT &#x7D4C;&#x8DEF;&#x30A8;&#x30D5;&#x30A7;&#x30AF;&#x30BF;&#x30FC; CTNNB1 &#xFF08;&#x4EE5;&#x4E0B;&#x3001;WNT &#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;) &#x306E;&#x6D3B;&#x6027;&#x5316;&#x5909;&#x7570;&#x3092;&#x542B;&#x3080;&#x9AC4;&#x82BD;&#x816B;&#x306E;&#x5C0F;&#x8133;&#x5916;&#x80CC;&#x306E;&#x8133;&#x5E79;&#x306E;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x767A;&#x751F;&#x3059;&#x308B;&#x304C;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x30DE;&#x30FC;&#x30AD;&#x30F3;&#x30B0;&#x4EBA;&#x9593; WNT &#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x306E;&#x9AC4;&#x82BD;&#x816B;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x3088;&#x308A;&#x983B;&#x7E41;&#x306B;&#x83F1;&#x5F62;&#x4E0B;&#x5507; &#xFF08;LRL&#xFF09; &#x3068;&#x80DA;&#x80CC;&#x8133;&#x5E79;&#x306B;&#x83F1;&#x5F62;&#x306E;&#x4E0A;&#x90E8;&#x306E; (URL) &#x5507;&#x3057;&#x3001;&#x5C0F;&#x8133;&#x306E;&#x958B;&#x767A;&#x3088;&#x308A;&#x3082;&#x3053;&#x3068;&#x3092;&#x898B;&#x3044;&#x3060;&#x3057;&#x305F;&#x3002;&#x6CE3;&#x304B;&#x306A;&#x3044;&#x3067;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x816B;&#x760D;&#x5C0F;&#x8133;&#x534A;&#x7403;&#x5185;&#x306B;&#x4F4D;&#x7F6E;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x4E00;&#x65B9;&#x4EBA;&#x9593; WNT &#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x816B;&#x760D;&#x80CC;&#x8133;&#x5E79;&#x306B;&#x4FB5;&#x5165;&#x3059;&#x308B;&#x78C1;&#x6C17;&#x5171;&#x9CF4;&#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0; &#xFF08;MRI&#xFF09; &#x3068;&#x8853;&#x4E2D;&#x306E;&#x30EC;&#x30DD;&#x30FC;&#x30C8;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;Ctnnb1 &#x306E;&#x5909;&#x7570;&#x6D3B;&#x6027;&#x5316;&#x3001;&#x5C0F;&#x8133;&#x524D;&#x99C6;&#x7D30;&#x80DE;&#x6570;&#x306B;&#x307B;&#x3068;&#x3093;&#x3069;&#x5F71;&#x97FF;&#x3092;&#x4E0E;&#x3048;&#x306A;&#x304B;&#x3063;&#x305F;&#x304C;&#x7D30;&#x80DE;&#x306E;&#x7570;&#x5E38;&#x306A;&#x84C4;&#x7A4D; Zic1(+) &#x306E;&#x7570;&#x5E38;&#x5897;&#x6B96;&#x524D;&#x99C6;&#x7D30;&#x80DE;&#x304C;&#x542B;&#x307E;&#x308C;&#x3066;&#x305D;&#x306E;&#x80DA;&#x80CC;&#x5074;&#x306E;&#x8133;&#x5E79;&#x306E;&#x4E0A;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x75C5;&#x5909;&#x306F;&#x5909;&#x7570;&#x5927;&#x4EBA;&#x306E;&#x30DE;&#x30A6;&#x30B9;&#x3067;&#x6C38;&#x7D9A;&#x5316;;&#x307E;&#x305F;&#x3001;Tp53 &#x304C;&#x540C;&#x6642;&#x306B;&#x524A;&#x9664;&#x3055;&#x308C;&#x305F;&#x5834;&#x5408;&#x306E; 15 &#xFF05; &#x3067;&#x3001;&#x9AC4;&#x82BD;&#x306F; WNT &#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x9AC4;&#x82BD;&#x816B;&#x306E;&#x89E3;&#x5256;&#x5B66;&#x3068;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x30D7;&#x30ED;&#x30D5;&#x30A1;&#x30A4;&#x30EB;&#x3092;&#x518D;&#x8868;&#x73FE;&#x5F62;&#x816B;&#x306B;&#x9032;&#x6B69;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005; &#x306F;&#x6211;&#x3005; &#x306E;&#x77E5;&#x8B58;&#x306B;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x9AC4;&#x82BD;&#x816B;&#x306E;&#x7570;&#x306A;&#x308B;&#x7D30;&#x80DE;&#x8D77;&#x6E90;&#x304C;&#x3042;&#x308B;&#x3053;&#x3068;&#x6700;&#x521D;&#x306E;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306E;&#x30C7;&#x30FC;&#x30BF;&#x6CE3;&#x304B;&#x306A;&#x3044;&#x3067;&#x3068; WNT &#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x306E;&#x9AC4;&#x82BD;&#x816B;&#x3068;&#x30DE;&#x30FC;&#x30AF;&#x306E;&#x5206;&#x5B50;&#x304A;&#x3088;&#x3073;&#x81E8;&#x5E8A;&#x306E;&#x9055;&#x3044;&#x306B;&#x3064;&#x3044;&#x3066;&#x8AAC;&#x660E;&#x3057;&#x3001;&#x4ECA;&#x5F8C;&#x306E;&#x7814;&#x7A76;&#x306E;&#x3053;&#x306E;&#x91CD;&#x8981;&#x306A;&#x5C0F;&#x5150;&#x304C;&#x3093;&#x306E;&#x6CBB;&#x7642;&#x306E;&#x305F;&#x3081;&#x306E;&#x6DF1;&#x9060;&#x306A;&#x542B;&#x610F;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;

&#x9AC4;&#x82BD;&#x816B;&#x306E;&#x30B5;&#x30D6;&#x30BF;&#x30A4;&#x30D7;&#x306B;&#x306F;&#x660E;&#x77AD;&#x306A;&#x767A;&#x9054;&#x7684;&#x8D77;&#x6E90;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;

Medulloblastoma &#xD568;&#xAED8; &#xAC00;&#xC7A5; &#xC77C;&#xBC18;&#xC801;&#xC778; &#xC545;&#xC131; &#xC5B4;&#xB9B0; &#xC2DC;&#xC808; &#xB1CC; &#xC885;&#xC591;&#xC744; &#xAD6C;&#xC131; &#xD558;&#xB294; &#xC784;&#xC0C1; &#xBD84;&#xC790;&#xB85C; &#xB2E4;&#xC591; &#xD55C; &#xC885;&#xC591; &#xD558;&#xC704; &#xCEEC;&#xB809;&#xC158;&#xC744; &#xD3EC;&#xD568; &#xD569;&#xB2C8;&#xB2E4;. &#xC18C;&#xB2C9; &#xD5E4; &#xC9C0; &#xD638; &#xADF8; &#xD1B5;&#xB85C; (&#xC774; &#xD558;, &#xC26C; &#xD558;&#xC704; &#xD615;&#xC2DD;)&#xC758; &#xD0C8; &#xC120; &#xD65C;&#xC131;&#xD654; &#xD6C4; &#xC57D; 25%&#xACFC; &#xB9BD; &#xC2E0;&#xACBD; &#xC804;&#xC870; &#xC138;&#xD3EC; (GNPCs)&#xC5D0;&#xC11C; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xC18C; &#xB1CC; &#xB0B4;&#xC5D0;&#xC11C; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xC774;&#xB7EC;&#xD55C; &#xC885;&#xC591; &#xC0DD;&#xAC01; &#xB41C;&#xB2E4;. &#xD558;&#xC704; &#xC720;&#xD615;&#xC744; &#xC54C; &#xC218; &#xC5C6;&#xB294; &#xB2E4;&#xB978; medulloblastoma &#xB4E4;&#xC774; &#xC6B4;&#xC804; &#xBCD1; &#xC801;&#xC778; &#xD504;&#xB85C;&#xC138;&#xC2A4; &#xC0C8;&#xB85C;&#xC6B4; &#xCE58;&#xB8CC; &#xD544;&#xC694;&#xD55C; &#xB300;&#xBD80;&#xBD84;&#xC758; &#xAC1C;&#xBC1C;&#xC744; &#xBC29;&#xD574;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC;&#xB294; WNT &#xACBD;&#xB85C; &#xC774;&#xD399;&#xD130; CTNNB1 (&#xC774; &#xD558;, WNT &#xD558;&#xC704;) &#xD65C;&#xC131;&#xD654; &#xBCC0;&#xC774; &#xD3EC;&#xD568; &#xD558;&#xB294; medulloblastoma&#xC758; &#xAC1C;&#xBCC4; &#xD558;&#xC704; &#xC18C; &#xB1CC; &#xBC16;&#xC5D0; &#xC11C; &#xC9C0; brainstem&#xC758; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xB9C8;&#xB984;&#xBAA8;&#xAF34; &#xC0C1;&#xB2E8;&#xC5D0; &#xB9BD; (URL) &#xBC0F; &#xC18C; &#xB1CC; &#xAC1C;&#xBC1C; &#xBCF4;&#xB2E4; &#xC720;&#xC804;&#xC790; &#xC778;&#xAC04;&#xC758; WNT &#xD558;&#xC704; medulloblastomas &#xB9C8;&#xD0B9; &#xB9C8;&#xB984;&#xBAA8;&#xAF34; &#xC544;&#xB7AB;&#xC785;&#xC220; (LRL) &#xBC0F; &#xBBF8; &#xBC1C;&#xB2EC; &#xB4F1; brainstem&#xC5D0;&#xB294; &#xD45C;&#xD604; &#xB354; &#xC790;&#xC8FC; &#xBC1C;&#xACAC;. &#xC790;&#xAE30; &#xACF5;&#xBA85; &#xC601;&#xC0C1; (MRI) &#xBC0F; &#xB0B4;&#xBD80; &#xBCF4;&#xACE0;&#xC11C; &#xC26C; &#xD558;&#xC704; &#xC885;&#xC591;&#xC740; &#xC18C; &#xB1CC; &#xBC18;&#xAD6C; &#xB0B4;&#xC5D0; &#xC788;&#xB294; &#xC778;&#xAC04;&#xC758; WNT &#xD558;&#xC704; &#xC885;&#xC591; &#xCE68;&#xD22C; &#xB4F1; brainstem &#xBCF4;&#xC600;&#xB2E4;. Ctnnb1&#xC5D0; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xD65C;&#xC131;&#xD654;&#xB294; &#xC18C; &#xB1CC;&#xC5D0; &#xC870;&#xC0C1; &#xC138;&#xD3EC; &#xC778;&#xAD6C;&#xC5D0; &#xAC70;&#xC758; &#xC601;&#xD5A5;&#xC744; &#xBBF8;&#xCE58;&#xC9C0; &#xD588;&#xB2E4; &#xD558;&#xC9C0;&#xB9CC; aberrantly &#xBC29;&#xC9C0;&#xD560; Zic1(+) &#xC804;&#xC870; &#xC138;&#xD3EC;&#xB97C; &#xD3EC;&#xD568; &#xD558;&#xB294; &#xBBF8; &#xBC1C;&#xB2EC; &#xB4F1; brainstem&#xC5D0; &#xC138;&#xD3EC;&#xC758; &#xBE44;&#xC815;&#xC0C1;&#xC801;&#xC778; &#xCD95;&#xC801;&#xC744; &#xBC1C;&#xC0DD;. &#xC774;&#xB7EC;&#xD55C; &#xBCD1; &#xBCC0; &#xC720;&#xC9C0; &#xBAA8;&#xB4E0; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC131;&#xC778; &#xC950;; &#xB610;&#xD55C;,&#xC758; Tp53 &#xB3D9;&#xC2DC;&#xC5D0; &#xC0AD;&#xC81C; &#xB41C; &#xACBD;&#xC6B0; 15%, &#xADF8;&#xB4E4;&#xC740; &#xC778;&#xAC04;&#xC758; WNT &#xD558;&#xC704; medulloblastoma&#xC758; &#xD574;&#xBD80;&#xD559; &#xBC0F; &#xC9C4; &#xC2DD; &#xD504;&#xB85C; &#xD30C;&#xC77C; recapitulated &#xC591;&#xC2DD; medulloblastomas &#xC9C4;&#xD589;. &#xC6B0;&#xB9AC; medulloblastoma&#xC758; &#xD558;&#xC704;&#xAC00; &#xB69C;&#xB837;&#xD55C; &#xC138;&#xD3EC; &#xAE30;&#xC6D0; &#xC6B0;&#xB9AC;&#xC758; &#xC9C0;&#xC2DD; &#xCCAB; &#xBC88;&#xC9F8; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xB370;&#xC774;&#xD130; &#xD45C;&#xC2DC; &#xB41C; &#xBD84;&#xC790;&#xC640; &#xC784;&#xC0C1; &#xCC28;&#xC774; &#xC26C; &#xBC0F; WNT &#xD558;&#xC704; medulloblastomas&#xC5D0; &#xB300; &#xD55C; &#xC124;&#xBA85;&#xC744; &#xC81C;&#xACF5; &#xD558; &#xACE0; &#xBBF8;&#xB798;&#xC758; &#xC5F0;&#xAD6C; &#xD558; &#xACE0;&#xC774; &#xC911;&#xC694; &#xD55C; &#xC18C;&#xC544; &#xC554; &#xCE58;&#xB8CC;&#xC5D0; &#xB300; &#xD55C; &#xAE4A;&#xC740; &#xC758;&#xBBF8;&#xB97C;&#xAC00;&#xC9C0;&#xACE0;.

Medulloblastoma&#xC758; &#xD558;&#xC704; &#xB69C;&#xB837;&#xD55C; &#xBC1C;&#xB2EC; &#xAE30;&#xC6D0; &#xD588;&#xC2B5;&#xB2C8;&#xB2E4;.

Meduloblastoma comprende una colecci&oacute;n de subtipos de tumor cl&iacute;nicamente y molecularmente diversas que abarcan el tumor m&aacute;s com&uacute;n del cerebro de malo de la infancia. Estos tumores se piensan para presentarse en el cerebelo, con aproximadamente 25% procedentes de c&eacute;lulas precursoras de neuronas del gr&aacute;nulo (GNPCs) despu&eacute;s de la activaci&oacute;n aberrante de la v&iacute;a de Sonic Hedgehog (en adelante, SHH subtipo). Los procesos patol&oacute;gicos que impulsan la heterogeneidad entre los otro meduloblastoma subtipos se desconocen, entorpeciendo el desarrollo de gran parte necesita nuevas terapias. Aqu&iacute; proporcionamos evidencia que un subtipo discreto de meduloblastoma que contiene mutaciones activadoras en el efector de la v&iacute;a WNT CTNNB1 (en adelante, subtipo WNT) surge fuera el cerebelo de las c&eacute;lulas de la m&eacute;dula oblonga dorsal. Encontramos que genes humanos meduloblastomas WNT-subtipo de la marca con m&aacute;s frecuencia se expresan en el labio r&oacute;mbico inferior (LRL) y embrionario dorsal del tronco encef&aacute;lico, que en la parte superior rombal (URL) del labio y el desarrollo de cerebelo. Informes intraoperatorias y la proyecci&oacute;n de imagen de resonancia magn&eacute;tica (MRI) demostraron que los tumores humanos de WNT-subtipo infiltran en el dorsal del tronco encef&aacute;lico, mientras que los tumores SHH-subtipo se encuentran dentro de los hemisferios cerebelosos. Activaci&oacute;n de mutaciones en Ctnnb1 tuvo poco impacto sobre las poblaciones de c&eacute;lulas progenitoras en el cerebelo, pero caus&oacute; la acumulaci&oacute;n anormal de c&eacute;lulas en el m&eacute;dula oblonga dorsal embrionario que incluy&oacute; aberrantemente proliferar las c&eacute;lulas del precursor de Zic1(+). Estas lesiones persistieron en todos los ratones mutantes para adultos; por otra parte, en el 15% de los casos en que simult&aacute;neamente se suprimi&oacute; el Tp53, progresaron a forma meduloblastomas que recapitulan los perfiles de expresi&oacute;n de anatom&iacute;a y gene de meduloblastoma de WNT-subtipo humano. Proporcionamos la primera evidencia, a nuestro conocimiento, que subtipos de meduloblastoma tienen or&iacute;genes distintos celulares. Nuestros datos proporcionan una explicaci&oacute;n de las marcadas diferencias moleculares y cl&iacute;nicas entre el subtipo de SHH y WNT meduloblastomas y tienen implicaciones profundas para futuras investigaciones y el tratamiento de este c&aacute;ncer infantil importante.

Subtipos de meduloblastoma tienen or&iacute;genes distintos de desarrollo.

It is generally believed that cerebellar granule neurons originate exclusively from granule neuron precursors (GNPs) in the external germinal layer (EGL). Here we identified a rare population of neuronal progenitors in mouse developing cerebellum that expresses Nestin. Although Nestin is widely considered a marker for multipotent stem cells, these Nestin-expressing progenitors (NEPs) are committed to the granule neuron lineage. Unlike conventional GNPs, which reside in the outer EGL and proliferate extensively, NEPs reside in the deep part of the EGL and are quiescent. Expression profiling revealed that NEPs are distinct from GNPs and, in particular, express markedly reduced levels of genes associated with DNA repair. Consistent with this, upon aberrant activation of Sonic hedgehog (Shh) signaling, NEPs exhibited more severe genomic instability and gave rise to tumors more efficiently than GNPs. These studies revealed a previously unidentified progenitor for cerebellar granule neurons and a cell of origin for medulloblastoma.

Zeng-jie Yang

Cancer Biology Program,
Fox Chase Cancer Center,
Cancer Biology Program, Fox Chase Cancer Center

Temple University Health System

Isolation of Distinct Cell Populations from the Developing Cerebellum by Microdissection

Zeng-jie Yang

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Cancer Biology Program,Fox Chase Cancer Center,Cancer Biology Program, Fox Chase Cancer Center

Temple University Health System

Isolation of Distinct Cell Populations from the Developing Cerebellum by Microdissection

Cancer Biology Program,
Fox Chase Cancer Center,
Cancer Biology Program, Fox Chase Cancer Center