in-vivo-modeling-of-the-morbid-human-genome-using-danio-rerio

<em>In Vivo</em> Modeling of the Morbid Human Genome using <em>Danio rerio</em>

Here,  we present methods for the development of assays to query potentially clinically  significant nonsynonymous changes using in  vivo complementation ...

Duke University Medical Center

Homogeneous and complete trisomy 8 is extremely rare. With one recent neonatal exception, all reported cases have been mosaic, due to mitotic non-disjunction during early zygotic development. We report a case of chromosome 8 trisomy in a human embryo examined at Carnegie stage 11 (25 days post-fertilization). It presented severe cardiovascular and central nervous system malformations.

Cytogenetic and histological features of a human embryo with homogeneous chromosome 8 trisomy.

We describe two fetal cases of microphthalmia/anophthalmia, pulmonary agenesis, and diaphragmatic defect. This rare association is known as Matthew-Wood syndrome (MWS; MIM 601186) or by the acronym "PMD" (Pulmonary agenesis, Microphthalmia, Diaphragmatic defect). Fewer than ten pre- and perinatal diagnoses of Matthew-Wood syndrome have been described to date. The cause is unknown, and the mode of transmission remains unclear. Most cases have been reported as isolated and sporadic, although recurrence among sibs has been observed once. Our two cases both occurred in consanguineous families, further supporting autosomal recessive transmission. In addition, in one family at least one of the elder sibs presented an evocatively similar phenotype. The spatiotemporal expression pattern of the FGF10 and FGFR2 genes in human embryos and the reported phenotypes of knockout mice for these genes spurred us to examine their coding sequences in our two cases of MWS. While in our patients, no causative sequence variations were identified in FGF10 or FGFR2, this cognate ligand-receptor pair and its downstream effectors remain functional candidates for MWS and similar associations of congenital ocular, diaphragmatic and pulmonary malformations.

Matthew-Wood syndrome: report of two new cases supporting autosomal recessive inheritance and exclusion of FGF10 and FGFR2.

Retinoic acid (RA) is a potent teratogen in all vertebrates when tight homeostatic controls on its endogenous dose, location, or timing are perturbed during early embryogenesis. STRA6 encodes an integral cell-membrane protein that favors RA uptake from soluble retinol-binding protein; its transcription is directly regulated by RA levels. Molecular analysis of STRA6 was undertaken in two human fetuses from consanguineous families we previously described with Matthew-Wood syndrome in a context of severe microphthalmia, pulmonary agenesis, bilateral diaphragmatic eventration, duodenal stenosis, pancreatic malformations, and intrauterine growth retardation. The fetuses had either a homozygous insertion/deletion in exon 2 or a homozygous insertion in exon 7 predicting a premature stop codon in STRA6 transcripts. Five other fetuses presenting at least one of the two major signs of clinical anophthalmia or pulmonary hypoplasia with at least one of the two associated signs of diaphragmatic closure defect or cardiopathy had no STRA6 mutations. These findings suggest a molecular basis for the prenatal manifestations of Matthew-Wood syndrome and suggest that phenotypic overlap with other associations may be due to genetic heterogeneity of elements common to the RA- and fibroblast growth factor-signaling cascades.

Matthew-Wood syndrome is caused by truncating mutations in the retinol-binding protein receptor gene STRA6.

&#x5C0F;&#x8111;&#x773C;&#x888B;&#x80BE;&#x7EFC;&#x5408;&#x5F81; (&#x9EA6;&#x5B50;)&#xFF0C;&#x4E5F;&#x79F0;&#x4E3A;&#x71C3;&#x6599;&#x7EFC;&#x5408;&#x5F81; B &#x578B;&#x548C;&#x6885;&#x514B;&#x5C14; (MKS) &#x7EFC;&#x5408;&#x5F81;&#x5C5E;&#x4E8E;&#x53D1;&#x80B2;&#x7684;&#x5E38;&#x67D3;&#x8272;&#x4F53;&#x663E;&#x6027;&#x9690;&#x6027;&#x969C;&#x788D;&#x539F;&#x53D1;&#x6027;&#x7EA4;&#x6BDB;&#x529F;&#x80FD;&#x969C;&#x788D;&#x4E0E;&#x76F8;&#x5173;&#x8054;&#x7684;&#x7EC4;&#x3002;&#x4F7F;&#x7528;&#x5355;&#x6838;&#x82F7;&#x9178;&#x591A;&#x6001;&#x6027;&#x7684;&#x6620;&#x5C04;&#xFF0C;&#x6211;&#x4EEC;&#x786E;&#x5B9A;&#x4E86;&#x5F02;&#x4E49;&#x548C;&#x622A;&#x65AD;&#x7A81;&#x53D8;&#x7684; RPGRIP1L (KIAA1005) &#x5728;&#x9EA6;&#x5B50;&#x548C; MKS&#xFF0C;&#x548C;&#x6211;&#x4EEC;&#x663E;&#x793A;&#x8BE5;&#x706D;&#x6D3B;&#x9F20;&#x6807; ortholog Rpgrip1l (Ftm) &#x7EAA;&#x8981;&#x7684;&#x9EA6;&#x5B50;&#x548C; MKS &#x7684;&#x8111;&#x3001; &#x80BE;&#x548C;&#x809D;&#x7684;&#x7F3A;&#x9677;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6211;&#x4EEC;&#x663E;&#x793A; RPGRIP1L colocalizes &#x5728;&#x57FA;&#x4F53;&#x548C;&#x4E2D;&#x5FC3;&#x4F53;&#x4E0E; NPHP6 &#x548C; NPHP4&#xFF0C;&#x5DF2;&#x77E5; MKS&#x3001; &#x9EA6;&#x5B50;&#x548C; nephronophthisis &#xFF08;&#x76F8;&#x5173;&#x7684;&#x80BE;&#x529F;&#x80FD;&#x969C;&#x788D;&#x548C; ciliopathy&#xFF09; &#x76F8;&#x5173;&#x57FA;&#x56E0;&#x86CB;&#x767D;&#x4EA7;&#x7269;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x5728;&#x9EA6;&#x5B50;&#x4E2A;&#x4EBA;&#x53D1;&#x73B0;&#x7684; RPGRIP1L &#x9519;&#x4E49;&#x7A81;&#x53D8;&#x964D;&#x4F4E;&#x4E86; RPGRIP1L &#x548C; nephrocystin-4 &#x4E4B;&#x95F4;&#x7684;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x663E;&#x793A; RPGRIP1L &#x7684;&#x7A81;&#x53D8;&#x53EF;&#x80FD;&#x4F1A;&#x5BFC;&#x81F4;&#x591A;&#x5668;&#x5B98;&#x8868;&#x578B;&#x5F02;&#x5E38;&#x9EA6;&#x5B50;&#x6216; MKS &#x4E2D;, &#x627E;&#x5230;&#xFF0C;&#x56E0;&#x6B64;&#x6784;&#x6210;&#x4E00;&#x4E2A;&#x7EDF;&#x4E00;&#x4F53;&#x7684;&#x76F8;&#x540C;&#x57FA;&#x7840;&#x969C;&#x788D;&#x3002;

&#x776B;&#x57FA;&#x56E0; RPGRIP1L &#x662F;&#x5C0F;&#x8111;&#x773C;&#x888B;&#x80BE;&#x7EFC;&#x5408;&#x5F81; &#xFF08;&#x71C3;&#x6599;&#x7EFC;&#x5408;&#x75C7; B &#x578B;&#xFF09; &#x548C;&#x6885;&#x514B;&#x5C14;&#x7EFC;&#x5408;&#x5F81;&#x7684;&#x7A81;&#x53D8;&#x3002;

Cerebello-oculo-renal syndrome (CORS), also called Joubert syndrome type B, and Meckel (MKS) syndrome belong to the group of developmental autosomal recessive disorders that are associated with primary cilium dysfunction. Using SNP mapping, we identified missense and truncating mutations in RPGRIP1L (KIAA1005) in both CORS and MKS, and we show that inactivation of the mouse ortholog Rpgrip1l (Ftm) recapitulates the cerebral, renal and hepatic defects of CORS and MKS. In addition, we show that RPGRIP1L colocalizes at the basal body and centrosomes with the protein products of both NPHP6 and NPHP4, known genes associated with MKS, CORS and nephronophthisis (a related renal disorder and ciliopathy). In addition, the RPGRIP1L missense mutations found in CORS individuals diminishes the interaction between RPGRIP1L and nephrocystin-4. Our findings show that mutations in RPGRIP1L can cause the multiorgan phenotypic abnormalities found in CORS or MKS, which therefore represent a continuum of the same underlying disorder.

The ciliary gene RPGRIP1L is mutated in cerebello-oculo-renal syndrome (Joubert syndrome type B) and Meckel syndrome.

C&#xE9;r&#xE9;bello-oculo-r&#xE9;nale (syndrome SCRO), &#xE9;galement appel&#xE9; syndrome de Joubert de type B, et Meckel (MKS) le syndrome appartiennent au groupe des maladies autosomiques r&#xE9;cessives de d&#xE9;veloppement qui sont associ&#xE9;s &#xE0; un dysfonctionnement cil primaire. Utilisation de la cartographie SNP, nous avons identifi&#xE9; des mutations faux-sens et tronquer dans RPGRIP1L (KIAA1005) dans les deux SCRO et MKS, et nous montrons que l&#x26;#39;inactivation du Rpgrip1l souris orthologue (FTM) r&#xE9;capitule les d&#xE9;fauts c&#xE9;r&#xE9;brale, r&#xE9;nale et h&#xE9;patique de la SCRO et MKS. En outre, nous montrons que RPGRIP1L colocalise &#xE0; la basale du corps et de centrosomes avec les produits prot&#xE9;iques de deux NPHP6 et NPHP4, les g&#xE8;nes connus li&#xE9;s &#xE0; MKS, la SCRO et n&#xE9;phronophtise (un trouble apparent&#xE9; r&#xE9;nale et ciliopathie). En outre, les mutations faux-sens RPGRIP1L trouv&#xE9;s chez les individus SCRO diminue l&#x26;#39;interaction entre RPGRIP1L et nephrocystin-4. Nos r&#xE9;sultats montrent que des mutations dans RPGRIP1L peut causer des anomalies ph&#xE9;notypiques plusieurs organes pr&#xE9;sents dans la SCRO ou MKS, qui repr&#xE9;sentent donc un continuum de la maladie sous-jacente m&#xEA;me.

Le RPGRIP1L Gene ciliaire est mut&#xE9; dans c&#xE9;r&#xE9;bello-oculo-r&#xE9;nale Syndrome (syndrome de Joubert de type B) et le syndrome de Meckel

Cerebello-Oculo-renale Syndrom (CORS), auch genannt Joubert-Syndrom Typ B und Meckel (MKS)-Syndrom geh&ouml;ren zur Gruppe der autosomal rezessiv Entwicklungsst&ouml;rungen, die prim&auml;re Zilie Dysfunktion zugeordnet sind. Mit SNP-Zuordnung, wir Missense und das K&uuml;rzen von Mutationen im RPGRIP1L identifiziert (KIAA1005) in CORS und MKS, und wir zeigen, dass Inaktivierung von der Maus Ortholog Rpgrip1l (Ftm) rekapituliert die zerebrale, renale und hepatische defekte CORS und MKS. Dar&uuml;ber hinaus zeigen wir, dass RPGRIP1L am basalen K&ouml;rper und Centrosomes mit der Protein-Produkte von NPHP6 und NPHP4, bekannt von Genen mit MKS, CORS und Nephronophthise (Verwandte renale St&ouml;rung und Ziliopathie,) colocalizes. Dar&uuml;ber hinaus die RPGRIP1L Missense-Mutationen gefunden in CORS Einzelpersonen verringert die Interaktion zwischen RPGRIP1L und Nephrocystin-4. Unsere Ergebnisse zeigen, dass Mutationen im RPGRIP1L die Syndrom ph&auml;notypischen Anomalien gefunden in CORS oder MKS, verursachen k&ouml;nnen, die daher ein Kontinuum von der gleichen zugrunde liegenden Erkrankung darstellen.

Cerebello-Oculo-renale Syndrom (Joubert-Syndrom Typ B) und Meckel-Syndrom ist das Zilien gen RPGRIP1L mutiert.

Sindrome cerebello-oculo-renale (CORS), chiamata anche sindrome di Joubert tipo B e la sindrome di Meckel (MKS) appartengono al gruppo di sviluppo malattie autosomiche recessive che sono associati con disfunzione cilio primario. Utilizzando il mapping del SNP, abbiamo identificato missenso e troncamento di mutazioni in RPGRIP1L (KIAA1005) in CORS e MKS, e ci mostrano che l&#39;inattivazione del mouse ortholog Rpgrip1l (Ftm) ricapitola i difetti cerebrali, renali ed epatici di CORS e MKS. Inoltre, mostriamo che RPGRIP1L colocalizza al basale e cntrosome con i prodotti della proteina di NPHP6 e di NPHP4, noti geni associati MKS, CORS e nefronoftisi (un disturbo renale correlata e Ciliopatia). Inoltre, le mutazioni missenso RPGRIP1L trovate in individui CORS diminuisce l&#39;interazione tra RPGRIP1L e nephrocystin-4. I nostri risultati mostrano che le mutazioni in RPGRIP1L possono causare le anomalie fenotipiche multiorgano trovate in CORS o MKS, che pertanto rappresentano un continuum della stessa patologia sottostante.

Il gene ciliare RPGRIP1L &egrave; mutato nella sindrome cerebello-oculo-renale (sindrome di Joubert tipo B) e la sindrome di Meckel.

&#x5C0F;&#x8133;&#x3001;&#x773C;&#x7403;&#x3001;&#x814E;&#x75C7;&#x5019;&#x7FA4;&#xFF08;CORS&#xFF09;&#x3082;&#x30B8;&#x30E5;&#x30D9;&#x30FC;&#x30EB;&#x75C7;&#x5019;&#x7FA4;B&#x578B;&#x3068;&#x547C;&#x3070;&#x308C;&#x3001;&#x30E1;&#x30C3;&#x30B1;&#x30EB;&#xFF08;MKS&#xFF09;&#x75C7;&#x5019;&#x7FA4;&#x306F;&#x3001;&#x4E00;&#x6B21;&#x7E4A;&#x6BDB;&#x306E;&#x6A5F;&#x80FD;&#x969C;&#x5BB3;&#x306B;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x767A;&#x9054;&#x5E38;&#x67D3;&#x8272;&#x4F53;&#x52A3;&#x6027;&#x907A;&#x4F1D;&#x6027;&#x75BE;&#x60A3;&#x306E;&#x30B0;&#x30EB;&#x30FC;&#x30D7;&#x306B;&#x5C5E;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002; SNP&#x30DE;&#x30C3;&#x30D4;&#x30F3;&#x30B0;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x3001;&#x6211;&#x3005;&#x306F;CORS&#x3068;MKS&#x306E;&#x4E21;&#x65B9;&#x3067;RPGRIP1L&#xFF08;KIAA1005&#xFF09;&#x306E;&#x30DF;&#x30B9;&#x30BB;&#x30F3;&#x30B9;&#x3068;&#x5207;&#x308A;&#x6368;&#x3066;&#x308B;&#x306E;&#x5909;&#x7570;&#x3092;&#x540C;&#x5B9A;&#x3057;&#x3001;&#x6211;&#x3005;&#x306F;&#x3001;&#x30DE;&#x30A6;&#x30B9;&#x306E;&#x30AA;&#x30EB;&#x30BD;&#x30ED;&#x30B0;Rpgrip1l&#xFF08;FTM&#xFF09;&#x3092;&#x7E70;&#x308A;&#x8FD4;&#x3059;&#x3068;&#x3044;&#x3046;CORS&#x3068;MKS&#x306E;&#x8133;&#x3001;&#x814E;&#x81D3;&#x304A;&#x3088;&#x3073;&#x809D;&#x81D3;&#x306E;&#x6B20;&#x9665;&#x305D;&#x306E;&#x4E0D;&#x6D3B;&#x6027;&#x5316;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x6211;&#x3005;&#x306F;RPGRIP1L&#x304C;NPHP6&#x3068;NPHP4&#x4E21;&#x65B9;&#x306E;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x7523;&#x7269;&#x3001;MKS&#x3001;CORS&#x3068;&#x30CD;&#x30D5;&#x30ED;&#x30F3;&#xFF08;&#x95A2;&#x9023;&#x3059;&#x308B;&#x814E;&#x969C;&#x5BB3;&#x3068;ciliopathy&#xFF09;&#x306B;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x65E2;&#x77E5;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x3068;&#x57FA;&#x790E;&#x4F53;&#x6E29;&#x3068;&#x4E2D;&#x5FC3;&#x4F53;&#x306B;&#x5C40;&#x5728;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;CORS&#x306E;&#x500B;&#x4F53;&#x306B;&#x898B;&#x3089;&#x308C;&#x308B;RPGRIP1L&#x30DF;&#x30B9;&#x30BB;&#x30F3;&#x30B9;&#x5909;&#x7570;&#x306F;RPGRIP1L&#x3068;nephrocystin-4&#x3068;&#x306E;&#x9593;&#x306E;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x3092;&#x6E1B;&#x5C11;&#x3055;&#x305B;&#x308B;&#x3002;&#x4ECA;&#x56DE;&#x306E;&#x77E5;&#x898B;&#x306F;&#x3001;RPGRIP1L&#x306E;&#x5909;&#x7570;&#x306F;&#x3001;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;&#x540C;&#x3058;&#x57FA;&#x790E;&#x75BE;&#x60A3;&#x306E;&#x9023;&#x7D9A;&#x3092;&#x8868;&#x3059;CORS&#x307E;&#x305F;&#x306F;MKS&#x3001;&#x306B;&#x898B;&#x3089;&#x308C;&#x308B;&#x81D3;&#x5668;&#x306E;&#x8868;&#x73FE;&#x578B;&#x306E;&#x7570;&#x5E38;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#x6BDB;&#x69D8;&#x4F53;&#x907A;&#x4F1D;&#x5B50;RPGRIP1L&#x306F;&#x3001;&#x5C0F;&#x8133;&#x3001;&#x773C;&#x7403;&#x3001;&#x814E;&#x75C7;&#x5019;&#x7FA4;&#xFF08;&#x30B8;&#x30E5;&#x30D9;&#x30FC;&#x30EB;&#x75C7;&#x5019;&#x7FA4;&#x30BF;&#x30A4;&#x30D7;B&#xFF09;&#x3068;&#x30E1;&#x30C3;&#x30B1;&#x30EB;&#x75C7;&#x5019;&#x7FA4;&#x3067;&#x5909;&#x7570;&#x3057;&#x3066;&#x3044;&#x308B;

Cerebello-oculo-&#xC2E0;&#xC7A5; &#xC99D;&#xD6C4;&#xAD70; (&#xCF54;&#xC2A4;), B &#xD615; Joubert &#xC99D;&#xD6C4;&#xAD70; &#xBC0F; Meckel (MKS) &#xC99D;&#xD6C4;&#xAD70;&#xC774; &#xB77C;&#xACE0;&#xB3C4; &#xAD00;&#xB828; &#xAE30;&#xBCF8; cilium &#xBD80;&#xC804; &#xB41C; &#xBC1C;&#xB2EC; &#xC0C1; &#xC5FC;&#xC0C9;&#xCCB4; &#xC5F4;&#xC131; &#xC9C8;&#xD658;&#xC758; &#xADF8;&#xB8F9;&#xC5D0; &#xC18D;&#xD55C;&#xB2E4;. Missense &#xBC0F; rpgrip1l&#xC5D0; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC790;&#xB978; &#xC2DD;&#xBCC4; SNP &#xB9E4;&#xD551; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; (KIAA1005) &#xCF54;&#xC2A4;&#xC640; MKS, &#xC6B0;&#xB9AC;&#xAC00; &#xB9C8;&#xC6B0;&#xC2A4; ortholog rpgrip1l&#xC758; &#xADF8; &#xBE44;&#xD65C;&#xC131;&#xD654; (Ftm) &#xCF54;&#xC2A4;&#xC640; MKS&#xC758; &#xB1CC;, &#xC2E0;&#xC7A5; &#xBC0F; &#xAC04;&#xC7A5; &#xACB0;&#xD568; &#xBC1C;&#xC0DD; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xC6B0;&#xB9AC;&#xB294; RPGRIP1L &#xAE30;&#xCD08; &#xC2E0;&#xCCB4;&#xC640; NPHP6 &#xBC0F; NPHP4, &#xC54C;&#xB824;&#xC9C4; MKS, &#xCF54;&#xC2A4; &#xBC0F; nephronophthisis (&#xAD00;&#xB828;&#xB41C; &#xC2E0;&#xC7A5; &#xC7A5;&#xC560;, ciliopathy)&#xC640; &#xAD00;&#xB828; &#xB41C; &#xC720;&#xC804;&#xC790;&#xC758; &#xB2E8;&#xBC31;&#xC9C8; &#xC81C;&#xD488;&#xC744; centrosomes&#xC5D0;&#xC11C; colocalizes&#xC744; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xCF54;&#xC2A4; &#xAC1C;&#xC778;&#xC5D0; &#xC788;&#xB294; RPGRIP1L missense &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; RPGRIP1L nephrocystin 4 &#xC0AC;&#xC774;&#xC758; &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC744; &#xAC10;&#xC18C; &#xD55C;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xBC1C;&#xACAC; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; rpgrip1l&#xC5D0;&#xB294; multiorgan phenotypic &#xC774;&#xC0C1;&#xC774; &#xCF54;&#xC2A4; &#xB610;&#xB294; MKS&#xC5D0; &#xB530;&#xB77C;&#xC11C; &#xAC19;&#xC740; &#xADFC;&#xBCF8;&#xC801;&#xC778; &#xC7A5;&#xC560;&#xC758; &#xC5F0;&#xC18D;&#xCCB4;&#xB97C; &#xB300;&#xD45C; &#xD558;&#xB294; &#xBC1C;&#xC0DD;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;.

&#xC18D;&#xB208;&#xC379; &#xC720;&#xC804;&#xC790; RPGRIP1L cerebello-oculo-&#xC2E0;&#xC7A5; &#xC99D;&#xD6C4;&#xAD70; (Joubert &#xC99D;&#xD6C4;&#xAD70; B &#xD615;)&#xACFC; Meckel &#xC99D;&#xD6C4;&#xAD70;&#xC5D0;&#xC11C; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC774;&#xB2E4;.

Cerebelo-&#xF3;culo-renal, s&#xED;ndrome (CORS), tambi&#xE9;n llamado s&#xED;ndrome de Joubert tipo B, y Meckel (MKS) el s&#xED;ndrome de pertenecer al grupo de desarrollo los trastornos autos&#xF3;micos recesivos que est&#xE1;n asociados con la disfunci&#xF3;n cilio primario. Uso de mapas SNP, se identificaron mutaciones missense y truncar en RPGRIP1L (KIAA1005) en ambos CORS y MKS, y nos muestran que la inactivaci&#xF3;n de la Rpgrip1l del rat&#xF3;n ortholog (FTM) recapitula los defectos cerebrales, renales y hep&#xE1;ticas de CORS y MKS. Adem&#xE1;s, nos muestran que RPGRIP1L colocalizes en la basal del cuerpo y los centrosomas con los productos de prote&#xED;na de ambos NPHP6 y los genes NPHP4, conocidos relacionados con MKS, CORS y la tuberculosis renal (un trastorno relacionado renal y ciliopathy). Adem&#xE1;s, las mutaciones missense RPGRIP1L encontrados en individuos CORS disminuye la interacci&#xF3;n entre RPGRIP1L y nephrocystin 4-. Nuestros hallazgos muestran que las mutaciones en RPGRIP1L puede causar las anormalidades fenot&#xED;picas multiorg&#xE1;nicas que se encuentran en CORS o MKS, que por lo tanto, representan un continuo de la enfermedad subyacente.

El RPGRIP1L Gene ciliar se encuentra mutado en cerebelo-&#xF3;culo-renal, s&#xED;ndrome (s&#xED;ndrome de Joubert Tipo B) y el s&#xED;ndrome de Meckel

Pierre Robin sequence (PRS) is an important subgroup of cleft palate. We report several lines of evidence for the existence of a 17q24 locus underlying PRS, including linkage analysis results, a clustering of translocation breakpoints 1.06-1.23 Mb upstream of SOX9, and microdeletions both approximately 1.5 Mb centromeric and approximately 1.5 Mb telomeric of SOX9. We have also identified a heterozygous point mutation in an evolutionarily conserved region of DNA with in vitro and in vivo features of a developmental enhancer. This enhancer is centromeric to the breakpoint cluster and maps within one of the microdeletion regions. The mutation abrogates the in vitro enhancer function and alters binding of the transcription factor MSX1 as compared to the wild-type sequence. In the developing mouse mandible, the 3-Mb region bounded by the microdeletions shows a regionally specific chromatin decompaction in cells expressing Sox9. Some cases of PRS may thus result from developmental misexpression of SOX9 due to disruption of very-long-range cis-regulatory elements.

Highly conserved non-coding elements on either side of SOX9 associated with Pierre Robin sequence.

Pitt-Hopkins syndrome is a severe congenital encephalopathy recently ascribed to de novo heterozygous TCF4 gene mutations. We report a series of 13 novel PHS cases with a TCF4 mutation and show that EEG, brain magnetic resonance imagain (MRI), and immunological investigations provide valuable additional clues to the diagnosis. We confirm a mutational hot spot in the basic domain of the E-protein. Functional studies illustrate that heterodimerisation of mutant TCF4 proteins with a tissue-specific transcription factor is less effective than that homodimerisation in a luciferase reporter assay. We also show that the TCF4 expression pattern in human embryonic development is widespread but not ubiquitous. In summary, we further delineate an underdiagnosed mental retardation syndrome, highlighting TCF4 function during development and facilitating diagnosis within the first year of life.

Mutational, functional, and expression studies of the TCF4 gene in Pitt-Hopkins syndrome.

Matthew-Wood, Spear, PDAC or MCOPS9 syndrome are alternative names used to refer to combinations of microphthalmia/anophthalmia, malformative cardiac defects, pulmonary dysgenesis, and diaphragmatic hernia. Recently, mutations in STRA6, encoding a membrane receptor for vitamin A-bearing plasma retinol binding protein, have been identified in such patients. We performed STRA6 molecular analysis in three fetuses and one child diagnosed with Matthew-Wood syndrome and in three siblings where two adult living brothers are affected with combinations of clinical anophthalmia, tetralogy of Fallot, and mental retardation. Among these patients, six novel mutations were identified, bringing the current total of known STRA6 mutations to seventeen. We extensively reviewed clinical data pertaining to all twenty-one reported patients with STRA6 mutations (the seven of this report and fourteen described elsewhere) and discuss additional features that may be part of the syndrome. The clinical spectrum associated with STRA6 deficiency is even more variable than initially described.

Phenotypic spectrum of STRA6 mutations: from Matthew-Wood syndrome to non-lethal anophthalmia.

FTO &#x662F;&#x5C5E;&#x4E8E; AlkB &#x76F8;&#x5173;&#x975E;&#x56E0;&#x5B50;&#x94C1;&#x548C; 2-oxoglutarate-&#x4F9D;&#x8D56;&#x7684;&#x53CC;&#x52A0;&#x6C27;&#x9176;&#x5BB6;&#x5EAD;&#x6838;&#x86CB;&#x767D;&#x8D28;&#x3002;&#x867D;&#x7136;&#x7B2C;&#x4E00;&#x4E2A;&#x5185;&#x542B;&#x5B50;&#x7684; FTO &#x57FA;&#x56E0;&#x591A;&#x6001;&#x6027;&#x4E0E;&#x80A5;&#x80D6;&#x76F8;&#x5173;&#x8054;&#xFF0C;FTO &#x7684;&#x751F;&#x7406;&#x4F5C;&#x7528;&#x4ECD;&#x7136;&#x662F;&#x672A;&#x77E5;&#x7684;&#x3002;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x663E;&#x793A;&#xFF0C;R316Q &#x7A81;&#x53D8;&#xFF0C;&#x706D;&#x6D3B; FTO &#x9176;&#x6D3B;&#x6027;&#xFF0C;&#x8D1F;&#x8D23;&#x4E3A;&#x5E38;&#x67D3;&#x8272;&#x4F53;&#x9690;&#x6027;&#x81F4;&#x6B7B;&#x7EFC;&#x5408;&#x5F81;&#x3002;&#x53D7;&#x5F71;&#x54CD;&#x7684;&#x4E3B;&#x9898;&#xFF0C;&#x8868;&#x660E;&#x4ECE;&#x4F53;&#x5916;&#x57F9;&#x517B;&#x7684;&#x76AE;&#x80A4;&#x6210;&#x7EA4;&#x7EF4;&#x7EC6;&#x80DE;&#x53D7;&#x635F;&#x6269;&#x6563;&#x548C;&#x52A0;&#x901F;&#x8870;&#x8001;&#x3002;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x8868;&#x660E; FTO &#x662F;&#x4EBA;&#x6743;&#x7684;&#x4E2D;&#x592E;&#x795E;&#x7ECF;&#x53CA;&#x5FC3;&#x8840;&#x7BA1;&#x7CFB;&#x7EDF;&#x7684;&#x6B63;&#x5E38;&#x53D1;&#x5C55;&#x7684;&#x5FC5;&#x8981;&#x6761;&#x4EF6;&#xFF0C;&#x5E76;&#x5EFA;&#x7ACB;&#x5728;&#x4EBA;&#x7C7B;&#x5927;&#x5BB6;&#x5EAD;&#x7684;&#x6210;&#x5458; AlkB &#x76F8;&#x5173;&#x53CC;&#x52A0;&#x6C27;&#x9176;&#x7684;&#x7A81;&#x53D8;&#x5BFC;&#x81F4;&#x4E25;&#x91CD; polymalformation &#x7EFC;&#x5408;&#x5F81;&#x3002;

&#x53CC;&#x52A0;&#x6C27;&#x9176;&#x7F16;&#x7801; FTO &#x57FA;&#x56E0;&#x529F;&#x80FD;&#x635F;&#x5931;&#x7684;&#x7A81;&#x53D8;&#x5BFC;&#x81F4;&#x4E25;&#x91CD;&#x53D1;&#x80B2;&#x8FDF;&#x7F13;&#x548C;&#x591A;&#x4E2A;&#x7578;&#x5F62;&#x3002;

FTO is a nuclear protein belonging to the AlkB-related non-haem iron- and 2-oxoglutarate-dependent dioxygenase family. Although polymorphisms within the first intron of the FTO gene have been associated with obesity, the physiological role of FTO remains unknown. Here we show that a R316Q mutation, inactivating FTO enzymatic activity, is responsible for an autosomal-recessive lethal syndrome. Cultured skin fibroblasts from affected subjects showed impaired proliferation and accelerated senescence. These findings indicate that FTO is essential for normal development of the central nervous and cardiovascular systems in human and establish that a mutation in a human member of the AlkB-related dioxygenase family results in a severe polymalformation syndrome.

Loss-of-function mutation in the dioxygenase-encoding FTO gene causes severe growth retardation and multiple malformations.

FTO est une prot&eacute;ine nucl&eacute;aire appartenant &agrave; la famille de dioxyg&eacute;nase d&eacute;pendante fer et 2-oxoglutarate AlkB li&eacute;s non h&eacute;minique. Bien que les polymorphismes dans le premier intron du g&egrave;ne FTO ont &eacute;t&eacute; associ&eacute;s &agrave; l&#39;ob&eacute;sit&eacute;, le r&ocirc;le physiologique de la FTO reste inconnu. Ici, nous montrons qu&#39;une mutation R316Q, d&eacute;sactivation d&#39;activit&eacute; enzymatique de la FTO, est responsable d&#39;un syndrome l&eacute;tal r&eacute;cessif autosomique. Fibroblastes cultiv&eacute;s de sujets atteints ont montr&eacute; avec facult&eacute;s affaiblies de prolif&eacute;ration et acc&eacute;l&egrave;re la s&eacute;nescence. Ces r&eacute;sultats indiquent que la FTO est essentielle au d&eacute;veloppement normal des syst&egrave;mes cardiovasculaires et nerveuses centrales chez l&#39;homme et &eacute;tablir qu&#39;une mutation dans un homme membre de la famille de li&eacute;s AlkB dioxyg&eacute;nase entra&icirc;ne un syndrome grave polymalformation.

Perte de fonction mutation dans le g&egrave;ne FTO dioxyg&eacute;nase-encodage provoque un retard de croissance s&eacute;v&egrave;re et des malformations multiples.

FTO ist ein nuklearer Protein aus der Alkbh1 im Zusammenhang mit nicht-Haem Eisen - und 2 Oxoglutarate-abh&auml;ngigen Dioxygenase Familie. Obgleich Polymorphien innerhalb der ersten Intron des FTO-Gens Adipositas zugeordnet wurden, ist die physiologische Rolle des FTO unbekannt. Hier zeigen wir, dass eine Mutation R316Q inaktivieren FTO enzymatische Aktivit&auml;t, ist verantwortlich f&uuml;r eine t&ouml;dliche autosomal-rezessive-Syndrom. Kultivierten Hautfibroblasten von betroffenen Probanden zeigten Vermehrung beeintr&auml;chtigt und beschleunigte Altern. Diese Befunde zeigen, dass FTO ist essentiell f&uuml;r die normale Entwicklung der zentralen Nerven- und Herz-Kreislauf-Systeme in der Human- und festzustellen, ob eine Mutation in einem menschlichen aus der Familie der Alkbh1-bezogenen Dioxygenase eine schwere Polymalformation-Syndrom f&uuml;hrt.

Loss of Function-Mutationen im Gen FTO Dioxygenase-Codierung verursacht schwere Wachstumsretardierung und multiple Fehlbildungen.

FTO &egrave; una proteina nucleare appartenente alla famiglia delle diossigenasi di ferro - e 2-ossoglutarato-dipendente Alkbh4-related non-eme. Sebbene polimorfismi all&#39;interno del primo introne del gene FTO sono stati associati con l&#39;obesit&agrave;, il ruolo fisiologico della FTO rimane sconosciuto. Qui ci mostrano che una mutazione R316Q, inattivazione enzimatica attivit&agrave; FTO, &egrave; responsabile di una sindrome letale autosomica recessiva. Fibroblasti cutanei da soggetti interessati ha mostrati ridotta proliferazione e accelerata senescenza. Questi risultati indicano che la FTO &egrave; essenziale per il normale sviluppo dei sistemi cardiovascolari e nervosi centrali umano e stabilire che una mutazione in un membro della famiglia Alkbh4-correlati diossigenasi umano provoca una sindrome grave polymalformation.

Mutazione di perdita di funzione del gene FTO codifica diossigenasi provoca grave ritardo della crescita e malformazioni multiple.

FTO &#x306F;&#x3001;AlkB &#x95A2;&#x9023;&#x975E;&#x30D8;&#x30E0;&#x9244;&#x304A;&#x3088;&#x3073; 2 &#x30AA;&#x30AD;&#x30BD;&#x30B0;&#x30EB;&#x30BF;&#x30EB;&#x9178;&#x4F9D;&#x5B58;&#x30B8;&#x30AA;&#x30AD;&#x30B7;&#x30B2;&#x30CA;&#x30FC;&#x30BC; &#x30D5;&#x30A1;&#x30DF;&#x30EA;&#x30FC;&#x306B;&#x5C5E;&#x3059;&#x308B;&#x6838;&#x86CB;&#x767D;&#x8CEA;&#x3067;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;FTO &#x306E;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x6700;&#x521D;&#x306E;&#x30A4;&#x30F3;&#x30C8;&#x30ED;&#x30F3;&#x5185;&#x306E;&#x591A;&#x5F62;&#x306F;&#x80A5;&#x6E80;&#x3068;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;FTO &#x306E;&#x751F;&#x7406;&#x7684;&#x5F79;&#x5272;&#x306F;&#x4E0D;&#x660E;&#x3067;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;R316Q &#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x3092;&#x793A;&#x3059; FTO &#x306E;&#x9175;&#x7D20;&#x6D3B;&#x6027;&#x3092;&#x4E0D;&#x6D3B;&#x6027;&#x5316;&#x3001;&#x5E38;&#x67D3;&#x8272;&#x4F53;&#x52A3;&#x6027;&#x81F4;&#x6B7B;&#x6027;&#x75C7;&#x5019;&#x7FA4;&#x306E;&#x305F;&#x3081;&#x306E;&#x8CAC;&#x4EFB;&#x3067;&#x3059;&#x3002;&#x5F71;&#x97FF;&#x3092;&#x53D7;&#x3051;&#x308B;&#x88AB;&#x9A13;&#x8005;&#x3092;&#x793A;&#x3057;&#x305F;&#x304B;&#x3089;&#x57F9;&#x990A;&#x76AE;&#x819A;&#x7DDA;&#x7DAD;&#x82BD;&#x7D30;&#x80DE;&#x5897;&#x6B96;&#x3092;&#x969C;&#x5BB3;&#x3057;&#x3001;&#x8001;&#x5316;&#x3092;&#x52A0;&#x901F;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x8ABF;&#x67FB;&#x7D50;&#x679C;&#x306F; FTO &#x4E2D;&#x592E;&#x306E;&#x795E;&#x7D4C;&#x3068;&#x5FC3;&#x8840;&#x7BA1;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x3067;&#x4EBA;&#x9593;&#x306E;&#x6B63;&#x5E38;&#x306A;&#x767A;&#x5C55;&#x306E;&#x305F;&#x3081;&#x306B;&#x4E0D;&#x53EF;&#x6B20;&#x3067;&#x3059; AlkB &#x95A2;&#x9023;&#x30B8;&#x30AA;&#x30AD;&#x30B7;&#x30B2;&#x30CA;&#x30FC;&#x30BC;&#x5BB6;&#x65CF;&#x306E;&#x4EBA;&#x9593;&#x30E1;&#x30F3;&#x30D0;&#x30FC;&#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x306F;&#x91CD;&#x75C7; polymalformation &#x75C7;&#x5019;&#x7FA4;&#x306B;&#x7D50;&#x679C;&#x3053;&#x3068;&#x3092;&#x78BA;&#x7ACB;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

&#x30B8;&#x30AA;&#x30AD;&#x30B7;&#x30B2;&#x30CA;&#x30FC;&#x30BC; &#x30A8;&#x30F3;&#x30B3;&#x30FC;&#x30C7;&#x30A3;&#x30F3;&#x30B0; FTO &#x306E;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x6A5F;&#x80FD;&#x55AA;&#x5931;&#x5909;&#x7570;&#x306F;&#x91CD;&#x75C7;&#x306E;&#x767A;&#x80B2;&#x3068;&#x591A;&#x767A;&#x6027;&#x5947;&#x5F62;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3057;&#x307E;&#x3059;&#x3002;

FTO AlkB &#xAD00;&#xB828; &#xB41C; &#xBE44;-haem &#xCCA0; &#xBC0F; 2-oxoglutarate-&#xC885;&#xC18D; dioxygenase &#xC81C;&#xD488;&#xAD70;&#xC5D0; &#xC18D;&#xD558;&#xB294; &#xD575; &#xB2E8;&#xBC31;&#xC9C8; &#xC774;&#xB2E4;. Polymorphisms FTO &#xC720;&#xC804;&#xC790;&#xC758; &#xCCAB; &#xBC88;&#xC9F8; intron &#xB0B4;&#xC5D0;&#xC11C; &#xBE44;&#xB9CC;&#xACFC; &#xC5F0;&#xAD00; &#xB41C;, &#xC788;&#xC9C0;&#xB9CC; FTO&#xC758; &#xC0DD;&#xB9AC; &#xC801; &#xC5ED;&#xD560;&#xC744; &#xC54C; &#xC218; &#xC5C6;&#xB294; &#xB0A8;&#xC544; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;&#xB294; R316Q &#xBCC0;&#xC774; &#xBCF4;&#xC5EC; &#xC0C1; &#xC5FC;&#xC0C9;&#xCCB4; &#xC5F4;&#xC131; &#xCE58;&#xBA85;&#xC801;&#xC778; &#xC99D;&#xD6C4;&#xAD70;&#xC5D0; &#xB300; &#xD55C; &#xCC45;&#xC784;&#xC740; FTO &#xD6A8;&#xC18C; &#xD65C;&#xB3D9;&#xC744; &#xBE44;&#xD65C;&#xC131;&#xD654; &#xD569;&#xB2C8;&#xB2E4;. &#xAD50;&#xC591;&#xB41C; &#xD53C;&#xBD80; &#xC12C;&#xC720; &#xC544; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xC601;&#xD5A5;&#xC744; &#xBC1B;&#xC740; &#xACFC;&#xBAA9; &#xD588;&#xB2E4; &#xD655;&#xC0B0; &#xC7A5;&#xC560; &#xBC0F; &#xB178;&#xD654; &#xAC00;&#xC18D;. &#xC774;&#xB7EC;&#xD55C; &#xC5F0;&#xAD6C; &#xACB0;&#xACFC; FTO &#xC778;&#xAC04;&#xC5D0;&#xC11C; &#xC911;&#xC559; &#xC2E0;&#xACBD; &#xBC0F; &#xC2EC;&#xC7A5; &#xD608;&#xAD00; &#xC2DC;&#xC2A4;&#xD15C;&#xC758; &#xC815;&#xC0C1;&#xC801;&#xC778; &#xBC1C;&#xC804;&#xC744; &#xC704;&#xD574; &#xD544;&#xC218;&#xC801; &#xC774;&#xB2E4; dioxygenase AlkB &#xAD00;&#xB828; &#xAC00;&#xC871;&#xC758; &#xC778;&#xAC04; &#xC77C;&#xC6D0;&#xC5D0; &#xBCC0;&#xC774; &#xC2EC;&#xD55C; polymalformation &#xC99D;&#xD6C4;&#xAD70;&#xC5D0; &#xACB0;&#xACFC; &#xC124;&#xC815; &#xD569;&#xB2C8;&#xB2E4; &#xB098;&#xD0C0;&#xB0C5;&#xB2C8;&#xB2E4;.

Dioxygenase &#xC778;&#xCF54;&#xB529; FTO &#xC720;&#xC804;&#xC790;&#xC5D0; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xAE30;&#xB2A5; &#xC190;&#xC2E4; &#xD558;&#xBA74; &#xC2EC;&#xD55C; &#xC131;&#xC7A5; &#xC9C0;&#xCCB4;&#xC640; &#xC5EC;&#xB7EC; &#xAE30;&#xD615;.

FTO es una prote&iacute;na nuclear perteneciente a la familia de dioxygenase de hierro 2-oxoglutarato dependientes y relacionados con AlkB no hemo. Aunque polimorfismos dentro del primer intr&oacute;n del gen FTO se han asociado con la obesidad, el papel fisiol&oacute;gico de FTO sigue siendo desconocido. Aqu&iacute; demostramos que una mutaci&oacute;n R316Q, inactivando la actividad enzim&aacute;tica de FTO, es responsable de un s&iacute;ndrome letal recesivo de un autosoma. Fibroblastos cultivados de la piel de los sujetos afectados mostraron alteraci&oacute;n proliferaci&oacute;n y aceleraron la senescencia. Estos resultados indican que la FTO es esencial para el desarrollo normal de los sistemas cardiovasculares y nerviosos centrales en humanos y establecer que una mutaci&oacute;n en un miembro humano de la familia relacionadas con AlkB dioxygenase da lugar a un s&iacute;ndrome polymalformation grave.

P&eacute;rdida de funci&oacute;n mutaci&oacute;n en el gen FTO dioxigenasa-codificaci&oacute;n causa retraso de crecimiento severo y malformaciones m&uacute;ltiples.

The LIM homeodomain gene Islet-1 (ISL1) encodes a transcription factor that has been associated with the multipotency of human cardiac progenitors, and in mice enables the correct deployment of second heart field (SHF) cells to become the myocardium of atria, right ventricle and outflow tract. Other markers have been identified that characterize subdomains of the SHF, such as the fibroblast growth factor Fgf10 in its anterior region. While functional evidence of its essential contribution has been demonstrated in many vertebrate species, SHF expression of Isl1 has been shown in only some models. We examined the relationship between human ISL1 and FGF10 within the embryonic time window during which the linear heart tube remodels into four chambers. ISL1 transcription demarcated an anatomical region supporting the conserved existence of a SHF in humans, and transcription factors of the GATA family were co-expressed therein. In conjunction, we identified a novel enhancer containing a highly conserved ISL1 consensus binding site within the FGF10 first intron. ChIP and EMSA demonstrated its direct occupation by ISL1. Transcription mediated by ISL1 from this FGF10 intronic element was enhanced by the presence of GATA4 and TBX20 cardiac transcription factors. Finally, transgenic mice confirmed that endogenous factors bound the human FGF10 intronic enhancer to drive reporter expression in the developing cardiac outflow tract. These findings highlight the interest of examining developmental regulatory networks directly in human tissues, when possible, to assess candidate non-coding regions that may be responsible for congenital malformations.

ISL1 directly regulates FGF10 transcription during human cardiac outflow formation.

Limb-girdle muscular dystrophy type 1D (LGMD1D) was linked to chromosome 7q36 over a decade ago, but its genetic cause has remained elusive. Here we studied nine LGMD-affected families from Finland, the United States and Italy and identified four dominant missense mutations leading to p.Phe93Leu or p.Phe89Ile changes in the ubiquitously expressed co-chaperone DNAJB6. Functional testing in vivo showed that the mutations have a dominant toxic effect mediated specifically by the cytoplasmic isoform of DNAJB6. In vitro studies demonstrated that the mutations increase the half-life of DNAJB6, extending this effect to the wild-type protein, and reduce its protective anti-aggregation effect. Further, we show that DNAJB6 interacts with members of the CASA complex, including the myofibrillar myopathy-causing protein BAG3. Our data identify the genetic cause of LGMD1D, suggest that its pathogenesis is mediated by defective chaperone function and highlight how mutations in a ubiquitously expressed gene can exert effects in a tissue-, isoform- and cellular compartment-specific manner.

Mutations affecting the cytoplasmic functions of the co-chaperone DNAJB6 cause limb-girdle muscular dystrophy.

Endoglin (CD105) is an endothelial-specific transforming growth factor Î² (TGF-Î²) coreceptor essential for angiogenesis and vascular homeostasis. Although endoglin dysfunction contributes to numerous vascular conditions, the mechanism of endoglin action remains poorly understood. Here we report a novel mechanism in which endoglin and GÎ±-interacting protein C-terminus-interacting protein (GIPC)-mediated trafficking of phosphatidylinositol 3-kinase (PI3K) regulates endothelial signaling and function. We demonstrate that endoglin interacts with the PI3K subunits p110Î± and p85 via GIPC to recruit and activate PI3K and Akt at the cell membrane. Opposing ligand-induced effects are observed in which TGF-Î²1 attenuates, whereas bone morphogenetic protein-9 enhances, endoglin/GIPC-mediated membrane scaffolding of PI3K and Akt to alter endothelial capillary tube stability in vitro. Moreover, we employ the first transgenic zebrafish model for endoglin to demonstrate that GIPC is a critical component of endoglin function during developmental angiogenesis in vivo. These studies define a novel non-Smad function for endoglin and GIPC in regulating endothelial cell function during angiogenesis.

Endoglin regulates PI3-kinase/Akt trafficking and signaling to alter endothelial capillary stability during angiogenesis.

Copy number variants (CNVs) are major contributors to genetic disorders. We have dissected a region of the 16p11.2 chromosome--which encompasses 29 genes--that confers susceptibility to neurocognitive defects when deleted or duplicated. Overexpression of each human transcript in zebrafish embryos identified KCTD13 as the sole message capable of inducing the microcephaly phenotype associated with the 16p11.2 duplication, whereas suppression of the same locus yielded the macrocephalic phenotype associated with the 16p11.2 deletion, capturing the mirror phenotypes of humans. Analyses of zebrafish and mouse embryos suggest that microcephaly is caused by decreased proliferation of neuronal progenitors with concomitant increase in apoptosis in the developing brain, whereas macrocephaly arises by increased proliferation and no changes in apoptosis. A role for KCTD13 dosage changes is consistent with autism in both a recently reported family with a reduced 16p11.2 deletion and a subject reported here with a complex 16p11.2 rearrangement involving de novo structural alteration of KCTD13. Our data suggest that KCTD13 is a major driver for the neurodevelopmental phenotypes associated with the 16p11.2 CNV, reinforce the idea that one or a small number of transcripts within a CNV can underpin clinical phenotypes, and offer an efficient route to identifying dosage-sensitive loci.

KCTD13 is a major driver of mirrored neuroanatomical phenotypes of the 16p11.2 copy number variant.

Nephronophthisis-related ciliopathies (NPHP-RC) are degenerative recessive diseases that affect kidney, retina, and brain. Genetic defects in NPHP gene products that localize to cilia and centrosomes defined them as "ciliopathies." However, disease mechanisms remain poorly understood. Here, we identify by whole-exome resequencing, mutations of MRE11, ZNF423, and CEP164 as causing NPHP-RC. All three genes function within the DNA damage response (DDR) pathway. We demonstrate that, upon induced DNA damage, the NPHP-RC proteins ZNF423, CEP164, and NPHP10 colocalize to nuclear foci positive for TIP60, known to activate ATM at sites of DNA damage. We show that knockdown of CEP164 or ZNF423 causes sensitivity to DNA damaging agents and that cep164 knockdown in zebrafish results in dysregulated DDR and an NPHP-RC phenotype. Our findings link degenerative diseases of the kidney and retina, disorders of increasing prevalence, to mechanisms of DDR.

Exome capture reveals ZNF423 and CEP164 mutations, linking renal ciliopathies to DNA damage response signaling.

Both the transforming growth factor Î² (TGF-Î²) and integrin signalling pathways have well-established roles in angiogenesis. However, how these pathways integrate to regulate angiogenesis is unknown. Here, we show that the extracellular matrix component, fibronectin, and its cellular receptor, Î±5Î²1 integrin, specifically increase TGF-Î²1- and BMP-9-induced Smad1/5/8 phosphorylation via the TGF-Î² superfamily receptors endoglin and activin-like kinase-1 (ALK1). Fibronectin and Î±5Î²1 integrin increase Smad1/5/8 signalling by promoting endoglin/ALK1 cell surface complex formation. In a reciprocal manner, TGF-Î²1 activates Î±5Î²1 integrin and downstream signalling to focal adhesion kinase (FAK) in an endoglin-dependent manner. Î±5Î²1 integrin and endoglin form a complex on the cell surface and co-internalize, with their internalization regulating Î±5Î²1 integrin activation and signalling. Functionally, endoglin-mediated fibronectin/Î±5Î²1 integrin and TGF-Î² pathway crosstalk alter the responses of endothelial cells to TGF-Î²1, switching TGF-Î²1 from a promoter to a suppressor of migration, inhibiting TGF-Î²1-mediated apoptosis to promote capillary stability, and partially mediating developmental angiogenesis in vivo. These studies provide a novel mechanism for the regulation of TGF-Î² superfamily signalling and endothelial function through crosstalk with integrin signalling pathways.

Endoglin mediates fibronectin/Î±5Î²1 integrin and TGF-Î² pathway crosstalk in endothelial cells.

Genomic rearrangements involving AUTS2 (7q11.22) are associated with autism and intellectual disability (ID), although evidence for causality is limited. By combining the results of diagnostic testing of 49,684 individuals, we identified 24 microdeletions that affect at least one exon of AUTS2, as well as one translocation and one inversion each with a breakpoint within the AUTS2 locus. Comparison of 17 well-characterized individuals enabled identification of a variable syndromic phenotype including ID, autism, short stature, microcephaly, cerebral palsy, and facial dysmorphisms. The dysmorphic features were more pronounced in persons with 3'AUTS2 deletions. This part of the gene is shown to encode a C-terminal isoform (with an alternative transcription start site) expressed in the human brain. Consistent with our genetic data, suppression of auts2 in zebrafish embryos caused microcephaly that could be rescued by either the full-length or the C-terminal isoform of AUTS2. Our observations demonstrate a causal role of AUTS2 in neurocognitive disorders, establish a hitherto unappreciated syndromic phenotype at this locus, and show how transcriptional complexity can underpin human pathology. The zebrafish model provides a valuable tool for investigating the etiology of AUTS2 syndrome and facilitating gene-function analysis in the future.

Exonic deletions in AUTS2 cause a syndromic form of intellectual disability and suggest a critical role for the C terminus.

Copy number variants (CNVs) represent a frequent type of lesion in human genetic disorders that typically affects numerous genes simultaneously. This has raised the challenge of understanding which genes within a CNV drive clinical phenotypes. Although CNVs can arise by multiple mechanisms, a subset is driven by local genomic architecture permissive to recombination events that can lead to both deletions and duplications. Phenotypic analyses of patients with such reciprocal CNVs have revealed instances in which the phenotype is either identical or mirrored; strikingly, molecular studies have shown that such phenotypes are often driven by reciprocal dosage defects of the same transcript. Here we explore how these observations can help the dissection of CNVs and inform the genetic architecture of CNV-induced disorders.

Article	Year
Cytogenetic and histological features of a human embryo with homogeneous chromosome 8 trisomy. Prenatal diagnosis\| PubMed ID: 17075794	2006
Matthew-Wood syndrome: report of two new cases supporting autosomal recessive inheritance and exclusion of FGF10 and FGFR2. American journal of medical genetics. Part A\| PubMed ID: 17236193	2007
Matthew-Wood syndrome is caused by truncating mutations in the retinol-binding protein receptor gene STRA6. American journal of human genetics\| PubMed ID: 17503335	2007
The ciliary gene RPGRIP1L is mutated in cerebello-oculo-renal syndrome (Joubert syndrome type B) and Meckel syndrome. Nature genetics\| PubMed ID: 17558409	2007
Highly conserved non-coding elements on either side of SOX9 associated with Pierre Robin sequence. Nature genetics\| PubMed ID: 19234473	2009
Mutational, functional, and expression studies of the TCF4 gene in Pitt-Hopkins syndrome. Human mutation\| PubMed ID: 19235238	2009
Phenotypic spectrum of STRA6 mutations: from Matthew-Wood syndrome to non-lethal anophthalmia. Human mutation\| PubMed ID: 19309693	2009
Loss-of-function mutation in the dioxygenase-encoding FTO gene causes severe growth retardation and multiple malformations. American journal of human genetics\| PubMed ID: 19559399	2009
ISL1 directly regulates FGF10 transcription during human cardiac outflow formation. PloS one\| PubMed ID: 22303449	2012
Mutations affecting the cytoplasmic functions of the co-chaperone DNAJB6 cause limb-girdle muscular dystrophy. Nature genetics\| PubMed ID: 22366786	2012
Endoglin regulates PI3-kinase/Akt trafficking and signaling to alter endothelial capillary stability during angiogenesis. Molecular biology of the cell\| PubMed ID: 22593212	2012
KCTD13 is a major driver of mirrored neuroanatomical phenotypes of the 16p11.2 copy number variant. Nature\| PubMed ID: 22596160	2012
Exome capture reveals ZNF423 and CEP164 mutations, linking renal ciliopathies to DNA damage response signaling. Cell\| PubMed ID: 22863007	2012
Endoglin mediates fibronectin/Î±5Î²1 integrin and TGF-Î² pathway crosstalk in endothelial cells. The EMBO journal\| PubMed ID: 22940691	2012
Exonic deletions in AUTS2 cause a syndromic form of intellectual disability and suggest a critical role for the C terminus. American journal of human genetics\| PubMed ID: 23332918	2013
Genetic architecture of reciprocal CNVs. Current opinion in genetics & development\| PubMed ID: 23747035	2013

Christelle Golzio

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