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1.
Gene ; 706: 62-68, 2019 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-31048069

RESUMO

Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is an autosomal dominant entity characterized by eyelid malformations and caused by mutations in the forkhead box L2 (FOXL2) gene. Clinical and genetic analyses of large cohorts of BPES patients from different ethnic origins are important for a better characterization of FOXL2 mutational landscape. The purpose of this study is to describe the phenotypic features and the causal FOXL2 variants in a Mexican cohort of BPES patients. A total of 12 individuals with typical facial findings were included. Clinical evaluation included palpebral measurements and levator function assessment. The complete coding sequence of FOXL2 was amplified by PCR and subsequently analyzed by Sanger sequencing. A total of 11 distinct FOXL2 pathogenic variants were identified in our cohort (molecular diagnostic rate of 92%), including 5 novel mutations. Our results broaden the BPES-related mutational spectrum and supports considerable FOXL2 allelic heterogeneity in our population.


Assuntos
Blefarofimose/genética , Proteína Forkhead Box L2/genética , Anormalidades da Pele/genética , Anormalidades Urogenitais/genética , Adolescente , Adulto , Blefarofimose/fisiopatologia , Criança , Pré-Escolar , Estudos de Coortes , Pálpebras/metabolismo , Feminino , Proteína Forkhead Box L2/fisiologia , Fatores de Transcrição Forkhead/genética , Humanos , Lactente , Recém-Nascido , Masculino , México , Pessoa de Meia-Idade , Mutação , Fenótipo , Anormalidades da Pele/fisiopatologia , Anormalidades Urogenitais/fisiopatologia
2.
Eur J Med Genet ; 62(7): 103668, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31077882

RESUMO

Blepharophimosis, Ptosis, and Epicanthus inversus Syndrome (BPES) is caused by autosomal dominant mutations in FOXL2. There are two forms of BPES: type I (with primary ovarian insufficiency (POI)) and type II (without POI). Data are presented from a large cohort of 177 BPES probands. Diagnostic testing identified a wide range of mutations in 119 mutation-positive patients (including 38 novel mutations). Although FOXL2 mutations are distributed throughout the gene, over 50% were frameshift mutations within a hotspot region of the gene that can be detected using a single primer pair to provide a cost-effective and rapid screening method. There was a significant proportion of de novo cases in this study, although in 7% there may be undetected parental mosaicism. There was an excess of female compared to male probands and a highly significant bias in the parental original of inherited mutations, with 20/21 found to be paternal in origin (95%). This could be because BPES in a female is more likely to come to clinical attention and because there is a generalised and more widespread clinical effect on fertility, in addition to the established association with POI. This study demonstrates the importance of cascade screening and provides new information on inheritance and parental mosaicism in BPES which will aid genetic counselling and accurate risk management.


Assuntos
Blefarofimose/genética , Proteína Forkhead Box L2/genética , Herança Paterna , Anormalidades da Pele/genética , Anormalidades Urogenitais/genética , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Mutação
4.
Mil Med Res ; 6(1): 4, 2019 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-30760330

RESUMO

BACKGROUND: Zinner syndrome represents a rare congenital malformation of the urinary tract. It comprises a constellation of Wolffian duct anomalies and is almost exclusively encountered as a classic triad of seminal vesicle cysts, ejaculatory duct obstruction and renal agenesis. Patients can be either asymptomatic or symptomatic. Recently, minimally invasive surgical techniques have emerged, superseding traditional surgery for select symptomatic cases. Our case highlights the finding of a rare clinical syndrome that was incidentally detected during a routine mass screening of military recruits in the Greek Armed Forces. CASE PRESENTATION: Herein, we present a case of a 19-year-old male who reported having a solitary right kidney when examined in a military training center of Northern Greece. No additional clinical information was available; thus, referral to a tertiary urology department for further investigation ensued. Imaging studies, namely, computed tomography and magnetic resonance imaging, revealed left renal aplasia, multiple left seminal vesicle cysts, and ejaculatory duct obstruction. Laboratory values and urinalysis were within normal range. Semen analysis was significant for cryptozoospermia. Our patient remained asymptomatic during the entire hospitalization. Long-term follow-up was recommended. Nevertheless, he declined further investigation and sought treatment in a private practice setting. CONCLUSIONS: This article aims to present the incidental diagnosis of a rare syndrome in a military setting. Population screening conducted in the armed forces permits the identification of undiagnosed diseases that warrant further investigation. To the best of our knowledge, this was the first report of Zinner syndrome in a military recruit and the second case cited of a Greek patient in the published literature. Regular follow-up is the key to timely intervention in conservatively managed cases.


Assuntos
Militares , Glândulas Seminais/anormalidades , Anormalidades Urogenitais/complicações , Anormalidades Congênitas/genética , Anormalidades Congênitas/fisiopatologia , Grécia , Humanos , Achados Incidentais , Rim/anormalidades , Rim/fisiopatologia , Masculino , Análise do Sêmen , Rim Único/complicações , Rim Único/fisiopatologia , Tomografia Computadorizada por Raios X/métodos , Anormalidades Urogenitais/diagnóstico , Anormalidades Urogenitais/etiologia , Anormalidades Urogenitais/genética , Anormalidades Urogenitais/fisiopatologia , Adulto Jovem
5.
DNA Cell Biol ; 38(3): 263-271, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30632787

RESUMO

Congenital anomalies of the kidney and urinary tract (CAKUT) are the leading cause of end-stage renal disease in children. Our group has discovered that Holliday Junction resolvase gene Gen1 is a potential candidate gene for CAKUT. Gen1 mutant mice showed CAKUT phenotypes similar to those observed in retinoic acid (RA)-deficient models. The expression of Raldh2, which encodes the key enzyme in RA synthesis, was reduced in Gen1 mutant metanephros through RNA sequencing. By real-time reverse transcription-PCR, the expression of both Raldh2 and downstream Ret was reduced in embryonic day (E) 11.5 Gen1 mutant ureters and E13.5 kidneys, and expression of RA receptor alpha was decreased in E13.5 Gen1 mutant ureters and kidneys. Further studies showed that all-trans retinoic acid (ATRA) rescued solitary kidney phenotype and improved ureteric branching; ATRA should be administered after ureteric budding to avoid increasing the incidence of ectopic budding in Gen1 mutants. Luciferase intensity of RA response element was lower in CHO-K1 cells transfected with Gen1 siRNA than in those transfected with scrambled RNA, and this inhibitory effect could be reversed by ATRA. These findings indicate that Gen1 mutation can result in renal malformation through RA signaling and Gen1-loss-induced CAKUT can be partly rescued by ATRA.


Assuntos
Resolvases de Junção Holliday/genética , Resolvases de Junção Holliday/metabolismo , Anormalidades Urogenitais/genética , Refluxo Vesicoureteral/genética , Animais , Células CHO , Cricetulus , Modelos Animais de Doenças , Rim/citologia , Rim/metabolismo , Camundongos , Mutação , Transdução de Sinais , Tretinoína/metabolismo , Sistema Urinário/metabolismo
6.
Mol Genet Genomics ; 294(2): 493-500, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30604070

RESUMO

Congenital anomalies of the kidney and urinary tract (CAKUT) are a wide range of congenital structural renal defects. CAKUT is the leading cause of chronic renal failure and end-stage renal disease in children. Studies in humans and animal models have confirmed the large genetic contribution to CAKUT. The previous evidence suggested that human TBX6 coding mutations might cause CAKUT via gene-dosage insufficiency. However, the potential involvement of TBX6 noncoding mutations in CAKUT remains to be elucidated. Here, we described DNA sequencing and copy-number analysis of TBX6 in 269 Chinese subjects with CAKUT. Interestingly, we identified two heterozygous noncoding variants of TBX6 in sporadic subjects with CAKUT: one is c.769-7delT, from a subject with duplex renal and collecting system, and the other is a 3' untranslated region (3'-UTR) variant (c.1392C>T) from a subject with unilateral renal hypoplasia. These two TBX6 noncoding variants are novel and extremely rare, respectively, in human populations archived in the ExAC database. The mini-gene splicing assay showed that the TBX6 c.769-7delT variant significantly reduced the splicing efficiency of TBX6 intron 5 when compared to the wild-type control. In this work, we identified a novel splicing variant of TBX6 in human CAKUT. Our experimental observations suggested that the TBX6 noncoding variant can affect gene expression and may potentially be involved in human CAKUT.


Assuntos
Sítios de Splice de RNA/genética , Análise de Sequência de DNA , Proteínas com Domínio T/genética , Anormalidades Urogenitais/genética , Refluxo Vesicoureteral/genética , Adolescente , Criança , Feminino , Humanos , Rim/fisiopatologia , Masculino , Mutação , Fenótipo , Anormalidades Urogenitais/fisiopatologia , Refluxo Vesicoureteral/fisiopatologia , Sequenciamento Completo do Exoma
7.
Nat Genet ; 51(1): 117-127, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30578417

RESUMO

Congenital anomalies of the kidney and urinary tract (CAKUT) are a major cause of pediatric kidney failure. We performed a genome-wide analysis of copy number variants (CNVs) in 2,824 cases and 21,498 controls. Affected individuals carried a significant burden of rare exonic (that is, affecting coding regions) CNVs and were enriched for known genomic disorders (GD). Kidney anomaly (KA) cases were most enriched for exonic CNVs, encompassing GD-CNVs and novel deletions; obstructive uropathy (OU) had a lower CNV burden and an intermediate prevalence of GD-CNVs; and vesicoureteral reflux (VUR) had the fewest GD-CNVs but was enriched for novel exonic CNVs, particularly duplications. Six loci (1q21, 4p16.1-p16.3, 16p11.2, 16p13.11, 17q12 and 22q11.2) accounted for 65% of patients with GD-CNVs. Deletions at 17q12, 4p16.1-p16.3 and 22q11.2 were specific for KA; the 16p11.2 locus showed extensive pleiotropy. Using a multidisciplinary approach, we identified TBX6 as a driver for the CAKUT subphenotypes in the 16p11.2 microdeletion syndrome.


Assuntos
Variações do Número de Cópias de DNA/genética , Rim/anormalidades , Sistema Urinário/anormalidades , Anormalidades Urogenitais/genética , Refluxo Vesicoureteral/genética , Deleção Cromossômica , Feminino , Predisposição Genética para Doença/genética , Estudo de Associação Genômica Ampla/métodos , Humanos , Masculino
8.
Proc Natl Acad Sci U S A ; 115(8): E1849-E1858, 2018 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-29432158

RESUMO

Genitourinary (GU) birth defects are among the most common yet least studied congenital malformations. Congenital anomalies of the kidney and urinary tract (CAKUTs) have high morbidity and mortality rates and account for ∼30% of structural birth defects. Copy number variation (CNV) mapping revealed that 16p11.2 is a hotspot for GU development. The only gene covered collectively by all of the mapped GU-patient CNVs was MYC-associated zinc finger transcription factor (MAZ), and MAZ CNV frequency is enriched in nonsyndromic GU-abnormal patients. Knockdown of MAZ in HEK293 cells results in differential expression of several WNT morphogens required for normal GU development, including Wnt11 and Wnt4. MAZ knockdown also prevents efficient transition into S phase, affects transcription of cell-cycle regulators, and abrogates growth of human embryonic kidney cells. Murine Maz is ubiquitously expressed, and a CRISPR-Cas9 mouse model of Maz deletion results in perinatal lethality with survival rates dependent on Maz copy number. Homozygous loss of Maz results in high penetrance of CAKUTs, and Maz is haploinsufficient for normal bladder development. MAZ, once thought to be a simple housekeeping gene, encodes a dosage-sensitive transcription factor that regulates urogenital development and contributes to both nonsyndromic congenital malformations of the GU tract as well as the 16p11.2 phenotype.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Fatores de Transcrição/metabolismo , Bexiga Urinária/anormalidades , Anormalidades Urogenitais/genética , Animais , Adesão Celular , Cromossomos Humanos Par 16 , Proteínas de Ligação a DNA/genética , Deleção de Genes , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , Masculino , Camundongos , Regiões Promotoras Genéticas , Interferência de RNA , RNA Interferente Pequeno , Fatores de Transcrição/genética , Transcrição Genética , Anormalidades Urogenitais/patologia
9.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 35(1): 96-99, 2018 Feb 10.
Artigo em Chinês | MEDLINE | ID: mdl-29419871

RESUMO

OBJECTIVE To assess the value of whole genome sequencing for the identification of de novo structural chromosomal abnormalities. METHODS Whole genome sequencing was utilized to analyze a boy with a peripheral blood karyotype of 46,XY,ins(3)(q21p13p21). The patient manifested with ocular abnormalities including blepharophimosis and ptosis. RESULTS Whole genome sequencing suggested a fragmentation of chromosome 3 (from position 55 473 257 to 78 341 929) has been inserted into between 136 876 730 to 138 643 831, and the breakpoints have occurred in the intergenic region. Meanwhile, there was a deletion between 138 643 831 and 138 694 476. This region contains FOXL2, a pathogenic gene associated with blepharophimosis-ptosis-epicanthus inversus syndrome. CONCLUSION De novo structural chromosomal abnormalities may be caused by novel breakpoints or microdeletion flanking the deletion region. To confirm its pathogenic nature, a mutation needs to be assessed at both genetic and genomic levels, for which whole genome sequencing is a good option.


Assuntos
Blefarofimose/genética , Deleção Cromossômica , Cromossomos Humanos Par 3/genética , Anormalidades da Pele/genética , Anormalidades Urogenitais/genética , Sequenciamento Completo do Genoma/métodos , Proteína Forkhead Box L2/genética , Deleção de Genes , Humanos , Lactente , Cariotipagem , Masculino
10.
Gene ; 650: 77-85, 2018 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-29410288

RESUMO

OBJECTIVE: The present study aimed to investigate the molecular mechanism underlying congenital obstructive nephropathy (CON). METHODS: The microarray dataset GSE70879 was downloaded from the Gene Expression Omnibus, including 3 kidney samples of megabladder mice and 4 control kidneys. Using this dataset, differentially expressed miRNAs (DEMs) were identified between the kidney samples from megabladder mice and controls, followed by identification of the target genes for these DEMs and construction of a DEM and target gene interaction network. Additionally, the target genes were subjected to Gene Ontology and pathway enrichment analyses, and were used for construction of a protein-protein interaction (PPI) network. Finally, regulatory networks were constructed to analyze transcription factors for the key miRNAs. RESULTS: From 17 DEMs identified between kidney samples of megabladder mice and controls, 3 key miRNAs were screened, including mmu-miR-150-5p, mmu-miR-374b-5p and mmu-miR-126a-5p. The regulatory networks identified vascular endothelial growth factor A (Vegfa) as the common target gene of mmu-miR-150-5p and five transcription factors, including nuclear receptor subfamily 4, group A, member 2 (Nr4a2), Jun dimerisation protein 2 (Jdp2), Kruppel-like factor 6 (Klf6), Neurexophilin-3 (Nxph3) and RNA binding motif protein 17 (Rbm17). The gene encoding phosphatase and tensin homolog (Pten) was found to be co-regulated by mmu-miR-374b-5p and high mobility group protein A1 (Hmga1), whereas the kirsten rat sarcoma viral oncogene (Kras) was identified as a common target gene of mmu-miR-126a-5p and paired box 6 (Pax6). CONCLUSIONS: In summary, the above-listed key miRNAs, transcription factors and key genes may be involved in the development of CON.


Assuntos
MicroRNAs/genética , Fatores de Transcrição/genética , Obstrução Ureteral/genética , Doenças da Bexiga Urinária/congênito , Doenças da Bexiga Urinária/genética , Anormalidades Urogenitais/genética , Animais , Modelos Animais de Doenças , Perfilação da Expressão Gênica , Ontologia Genética , Redes Reguladoras de Genes , Humanos , Camundongos , Análise em Microsséries , Mapas de Interação de Proteínas , Transdução de Sinais/genética , Obstrução Ureteral/congênito , Obstrução Ureteral/patologia , Doenças da Bexiga Urinária/patologia , Anormalidades Urogenitais/patologia
12.
Mol Genet Genomic Med ; 6(2): 261-267, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29378385

RESUMO

BACKGROUND: Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a malformation of the eyelids. Forkhead Box L2 (FOXL2) is the only gene known to be associated with BPES. METHODS: We identified two Han Chinese BPES families with premature ovarian insufficiency (POI). Sanger sequencing and in vitro functional analysis were performed to identify the genetic cause. RESULTS: Sanger sequencing identified two novel mutations (c.462_468del, c.988_989insG) in FOXL2, one in each family. The in vitro functional analysis confirmed that both novel mutations were associated with impaired transactivation of downstream genes. Specifically, the single-base insertion, c.988_989insG, led to subcellular mislocalization and aggregation of the encoded protein, which validated the hypothesis that the two novel FOXL2 mutations are deleterious and associated with POI in the two BPES families. CONCLUSION: The novel mutations identified in the present study will enhance the present knowledge of the mutation spectrum of FOXL2. The in vitro experiments provide further insights into the molecular mechanism by which the two new variants mediate disease pathogenesis and may contribute to elucidating the genotype-phenotype correlation between the two novel FOXL2 mutations and POI.


Assuntos
Blefarofimose/genética , Proteína Forkhead Box L2/genética , Insuficiência Ovariana Primária/genética , Anormalidades da Pele/genética , Anormalidades Urogenitais/genética , Adulto , Sequência de Bases/genética , Blefarofimose/complicações , Blefarofimose/metabolismo , China , Grupos Étnicos/genética , Pálpebras/anormalidades , Feminino , Proteína Forkhead Box L2/metabolismo , Fatores de Transcrição Forkhead/genética , Estudos de Associação Genética , Humanos , Linhagem , Insuficiência Ovariana Primária/complicações , Anormalidades da Pele/metabolismo , Anormalidades Urogenitais/metabolismo
13.
J Hum Genet ; 63(4): 529-532, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29362492

RESUMO

The diphthamide biosynthesis 1 (DPH1) gene encodes one of the essential components of the enzyme catalyzing the first step of diphthamide formation on eukaryotic elongation factor 2 (EEF2). Diphthamide is the posttranslationally modified histidine residue on EEF2 that promotes protein chain elongation in the ribosome. DPH1 defects result in a failure of protein synthesis involving EEF2, leading to growth defects, embryonic lethality, and cell death. In humans, DPH1 mutations cause developmental delay with a short stature, dysmorphic features, and sparse hair, and are inherited in an autosomal recessive manner (MIM#616901). To date, only two homozygous missense mutations in DPH1 (c.17T>A, p.Met6Lys and c.701T>C, p.Leu234Pro) have been reported. We used WES to identify novel compound heterozygous mutations in DPH1 (c.289delG, p.Glu97Lysfs*8 and c.491T>C, p.Leu164Pro) in a patient from a nonconsanguineous family presenting with intellectual disability, a short stature, craniofacial abnormalities, and external genital abnormalities. The clinical phenotype of all patients with DPH1 mutations, including the current patient, revealed core features, although the external genital anomaly was newly recognized in our case.


Assuntos
Obstrução das Vias Respiratórias/diagnóstico , Obstrução das Vias Respiratórias/genética , Heterozigoto , Antígenos de Histocompatibilidade Menor/genética , Mutação , Fenótipo , Proteínas Supressoras de Tumor/genética , Anormalidades Urogenitais/diagnóstico , Anormalidades Urogenitais/genética , Alelos , Substituição de Aminoácidos , Encéfalo/anormalidades , Encéfalo/diagnóstico por imagem , Análise Mutacional de DNA , Facies , Humanos , Recém-Nascido , Masculino , Linhagem , Síndrome
14.
PLoS One ; 13(1): e0191224, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29351342

RESUMO

Congenital anomalies of the kidney and urinary tract (CAKUT) are the most common cause (40-50%) of chronic kidney disease (CKD) in children. About 40 monogenic causes of CAKUT have so far been discovered. To date less than 20% of CAKUT cases can be explained by mutations in these 40 genes. To identify additional monogenic causes of CAKUT, we performed whole exome sequencing (WES) and homozygosity mapping (HM) in a patient with CAKUT from Indian origin and consanguineous descent. We identified a homozygous missense mutation (c.1336C>T, p.Arg446Cys) in the gene Von Willebrand factor A domain containing 2 (VWA2). With immunohistochemistry studies on kidneys of newborn (P1) mice, we show that Vwa2 and Fraser extracellular matrix complex subunit 1 (Fras1) co-localize in the nephrogenic zone of the renal cortex. We identified a pronounced expression of Vwa2 in the basement membrane of the ureteric bud (UB) and derivatives of the metanephric mesenchyme (MM). By applying in vitro assays, we demonstrate that the Arg446Cys mutation decreases translocation of monomeric VWA2 protein and increases translocation of aggregated VWA2 protein into the extracellular space. This is potentially due to the additional, unpaired cysteine residue in the mutated protein that is used for intermolecular disulfide bond formation. VWA2 is a known, direct interactor of FRAS1 of the Fraser-Complex (FC). FC-encoding genes and interacting proteins have previously been implicated in the pathogenesis of syndromic and/or isolated CAKUT phenotypes in humans. VWA2 therefore constitutes a very strong candidate in the search for novel CAKUT-causing genes. Our results from in vitro experiments indicate a dose-dependent neomorphic effect of the Arg446Cys homozygous mutation in VWA2.


Assuntos
Biomarcadores Tumorais/genética , Síndrome de Fraser/genética , Mutação de Sentido Incorreto , Anormalidades Urogenitais/genética , Refluxo Vesicoureteral/genética , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Animais Recém-Nascidos , Biomarcadores Tumorais/química , Criança , Consanguinidade , Sequência Conservada , Éxons , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Homozigoto , Humanos , Masculino , Camundongos , Modelos Animais , Modelos Moleculares , Linhagem , Homologia de Sequência de Aminoácidos , Sistema Urogenital/crescimento & desenvolvimento , Sistema Urogenital/metabolismo
15.
Cell Physiol Biochem ; 45(1): 203-211, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29339661

RESUMO

BACKGROUND/AIMS: Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a rare autosomal dominant disease caused by FOXL2 gene mutations, and it is clinically characterized by an eyelid malformation associated (type I) or not (type II) with premature ovarian failure (POF). Functional study of novel mutations is especially critical for female patients, as it may allow the prediction of infertility and early planning of an appropriate therapy. METHODS: A clinical and molecular genetic investigation was performed in all members of a Chinese family with BPES. Genomic DNA was extracted, and the FOXL2 coding region was sequenced. Subcellular localization was performed by confocal microscopy. Transactivation studies were performed by real-time PCR, dual luciferase reporter assays and electrophoretic mobility shift assays. RESULTS: A novel deletion mutation (C.634_641 del, CCCATGC) between the forkhead domain and the polyalanine domain was found, resulting in a frameshift mutation and a truncated protein. Functional studies showed a strong cytoplasmic mislocalization and abnormal transactivation activity, implying a type I kind mutation with a large chance of infertility. CONCLUSION: This study identifies that this mutation indicates the probability of developing into POF and shows the importance and necessity of early recognition of BPES type through mutation testing for female patients. Prompt personalized therapy and follow-up is of great clinical significance for female patients carrying this kind of mutation.


Assuntos
Blefarofimose/genética , Proteína Forkhead Box L2/genética , Anormalidades da Pele/genética , Anormalidades Urogenitais/genética , Sequência de Bases , Blefarofimose/patologia , Criança , Citoplasma/metabolismo , DNA/química , DNA/isolamento & purificação , DNA/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Feminino , Proteína Forkhead Box L2/metabolismo , Mutação da Fase de Leitura , Humanos , Masculino , Microscopia Confocal , Linhagem , Análise de Sequência de DNA , Deleção de Sequência , Anormalidades da Pele/patologia , Ativação Transcricional , Anormalidades Urogenitais/patologia
16.
Rev Chil Pediatr ; 89(6): 741-746, 2018 Dec.
Artigo em Espanhol | MEDLINE | ID: mdl-30725063

RESUMO

INTRODUCTION: Congenital anomalies of the kidney and urinary tract are caused by genetic alterations mostly unknown. Mutations in the gene that codes for hepatocyte nuclear factor 1B (HNF1B) are the most frequently described monogenic causes. Data are unknown in Chile and Latin America. OBJECTIVE: To determine the presence of variants of the HNF1B gene in Chilean children with conge nital anomalies of the kidney and/or the urinary tract and their clinical characteristics. PATIENTS AND METHOD: Descriptive study with children aged 10 months to 17 years, patients of the Calvo Mackenna Hospital Nephrology Unit, with cystic renal dysplasia, non cystic renal dysplasia/hypoplasia, horses hoe kidney between April and December 2016. HNF1B variants were determined by sequencing of exons 1, 2, 3 and 4 after DNA extraction and amplification. Restriction enzymes were used to define if the variants were homo or heterozygous. Direct family members of index cases were studied with sequencing of the affected exon. RESULTS: 32 patients were included, 43.75% males, median age 11 years. 65.6% of them had non-cystic renal dysplasia, 31.25% cystic renal dysplasia, and 3.15% hor seshoe kidney. In two patients (6.25%) the same heterozygous genetic variant was detected in exon 4, position 1027 (C1027T), not previously described. The study of relatives found the same variant in three out of five individuals, all without congenital nephro-urological anomalies. CONCLUSIONS: We confirmed the presence of a not previously described heterozygous genetic variant of the HNF1B gene. This work initiates the search for this type of mutations in our region which allows us to ap proach the knowledge of causality, determination of extrarenal involvement, and genetic counseling.


Assuntos
Fator 1-beta Nuclear de Hepatócito/genética , Nefropatias/genética , Anormalidades Urogenitais/genética , Doenças Urológicas/genética , Adolescente , Criança , Pré-Escolar , Chile , Feminino , Marcadores Genéticos , Predisposição Genética para Doença , Heterozigoto , Humanos , Lactente , Masculino , Mutação , Estudos Prospectivos
17.
Development ; 144(24): 4704-4719, 2017 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-29158444

RESUMO

Kidney development depends crucially on proper ureteric bud branching giving rise to the entire collecting duct system. The transcription factor HNF1B is required for the early steps of ureteric bud branching, yet the molecular and cellular events regulated by HNF1B are poorly understood. We report that specific removal of Hnf1b from the ureteric bud leads to defective cell-cell contacts and apicobasal polarity during the early branching events. High-resolution ex vivo imaging combined with a membranous fluorescent reporter strategy show decreased mutant cell rearrangements during mitosis-associated cell dispersal and severe epithelial disorganization. Molecular analysis reveals downregulation of Gdnf-Ret pathway components and suggests that HNF1B acts both upstream and downstream of Ret signaling by directly regulating Gfra1 and Etv5 Subsequently, Hnf1b deletion leads to massively mispatterned ureteric tree network, defective collecting duct differentiation and disrupted tissue architecture, which leads to cystogenesis. Consistently, mRNA-seq analysis shows that the most impacted genes encode intrinsic cell-membrane components with transporter activity. Our study uncovers a fundamental and recurring role of HNF1B in epithelial organization during early ureteric bud branching and in further patterning and differentiation of the collecting duct system in mouse.


Assuntos
Polaridade Celular/genética , Fator 1-beta Nuclear de Hepatócito/genética , Túbulos Renais Coletores/embriologia , Ureter/embriologia , Anormalidades Urogenitais/embriologia , Anormalidades Urogenitais/genética , Animais , Adesão Celular/genética , Células Cultivadas , Proteínas de Ligação a DNA/metabolismo , Regulação para Baixo/genética , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Fator 1-beta Nuclear de Hepatócito/metabolismo , Camundongos , Camundongos Knockout , Proteínas Nucleares/metabolismo , Técnicas de Cultura de Órgãos , Fator de Transcrição PAX2/biossíntese , Transdução de Sinais/genética , Fatores de Transcrição/metabolismo
18.
Am J Med Genet A ; 173(12): 3226-3230, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29088509

RESUMO

The STAR syndrome is a rare X-linked dominant developmental disorder caused by point mutations in the single FAM58A gene or deletions involving FAM58A and its flanking genes. The STAR phenotype is characterized by a rather homogeneous constellation of facial dysmorphisms and malformations summarized by its acronym, Syndactyly, Telecanthus, Anogenital, and Renal malformations. Here we describe a female patient with STAR syndrome and a 130 kb deletion at Xq28, including the FAM58A gene. She presented with cleft lip palate, omphalocele, and cerebral malformations not previously considered part of the phenotypic spectrum of this syndrome. She died at 6 weeks from respiratory failure.


Assuntos
Canal Anal/anormalidades , Fissura Palatina/genética , Ciclinas/genética , Hipertelorismo/genética , Rim/anormalidades , Sindactilia/genética , Dedos do Pé/anormalidades , Anormalidades Urogenitais/genética , Canal Anal/patologia , Bandeamento Cromossômico , Fissura Palatina/diagnóstico , Fissura Palatina/patologia , Evolução Fatal , Feminino , Humanos , Hipertelorismo/diagnóstico , Hipertelorismo/patologia , Recém-Nascido , Cariotipagem , Rim/patologia , Mutação Puntual , Sindactilia/diagnóstico , Sindactilia/patologia , Dedos do Pé/patologia , Anormalidades Urogenitais/diagnóstico , Anormalidades Urogenitais/patologia
19.
Am J Hum Genet ; 101(5): 803-814, 2017 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-29100091

RESUMO

Congenital anomalies of the kidney and urinary tract (CAKUT) constitute a major cause of chronic kidney disease in children and 20% of prenatally detected anomalies. CAKUT encompass a spectrum of developmental kidney defects, including renal agenesis, hypoplasia, and cystic and non-cystic dysplasia. More than 50 genes have been reported as mutated in CAKUT-affected case subjects. However, the pathophysiological mechanisms leading to bilateral kidney agenesis (BKA) remain largely elusive. Whole-exome or targeted exome sequencing of 183 unrelated familial and/or severe CAKUT-affected case subjects, including 54 fetuses with BKA, led to the identification of 16 heterozygous variants in GREB1L (growth regulation by estrogen in breast cancer 1-like), a gene reported as a target of retinoic acid signaling. Four loss-of-function and 12 damaging missense variants, 14 being absent from GnomAD, were identified. Twelve of them were present in familial or simplex BKA-affected case subjects. Female BKA-affected fetuses also displayed uterus agenesis. We demonstrated a significant association between GREB1L variants and BKA. By in situ hybridization, we showed expression of Greb1l in the nephrogenic zone in developing mouse kidney. We generated a Greb1l knock-out mouse model by CRISPR-Cas9. Analysis at E13.5 revealed lack of kidneys and genital tract anomalies in male and female Greb1l-/- embryos and a slight decrease in ureteric bud branching in Greb1l+/- embryos. We showed that Greb1l invalidation in mIMCD3 cells affected tubulomorphogenesis in 3D-collagen culture, a phenotype rescued by expression of the wild-type human protein. This demonstrates that GREB1L plays a major role in early metanephros and genital development in mice and humans.


Assuntos
Anormalidades Congênitas/genética , Nefropatias/congênito , Rim/anormalidades , Mutação/genética , Proteínas de Neoplasias/genética , Proteínas/genética , Animais , Criança , Exoma/genética , Feminino , Feto/anormalidades , Heterozigoto , Humanos , Nefropatias/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fenótipo , Sistema Urinário/anormalidades , Anormalidades Urogenitais/genética
20.
Am J Hum Genet ; 101(5): 789-802, 2017 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-29100090

RESUMO

Renal agenesis and hypodysplasia (RHD) are major causes of pediatric chronic kidney disease and are highly genetically heterogeneous. We conducted whole-exome sequencing in 202 case subjects with RHD and identified diagnostic mutations in genes known to be associated with RHD in 7/202 case subjects. In an additional affected individual with RHD and a congenital heart defect, we found a homozygous loss-of-function (LOF) variant in SLIT3, recapitulating phenotypes reported with Slit3 inactivation in the mouse. To identify genes associated with RHD, we performed an exome-wide association study with 195 unresolved case subjects and 6,905 control subjects. The top signal resided in GREB1L, a gene implicated previously in Hoxb1 and Shha signaling in zebrafish. The significance of the association, which was p = 2.0 × 10-5 for novel LOF, increased to p = 4.1 × 10-6 for LOF and deleterious missense variants combined, and augmented further after accounting for segregation and de novo inheritance of rare variants (joint p = 2.3 × 10-7). Finally, CRISPR/Cas9 disruption or knockdown of greb1l in zebrafish caused specific pronephric defects, which were rescued by wild-type human GREB1L mRNA, but not mRNA containing alleles identified in case subjects. Together, our study provides insight into the genetic landscape of kidney malformations in humans, presents multiple candidates, and identifies SLIT3 and GREB1L as genes implicated in the pathogenesis of RHD.


Assuntos
Anormalidades Congênitas/genética , Exoma/genética , Nefropatias/congênito , Rim/anormalidades , Mutação/genética , Proteínas de Neoplasias/genética , Alelos , Animais , Estudos de Casos e Controles , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Feminino , Heterogeneidade Genética , Estudo de Associação Genômica Ampla/métodos , Genótipo , Hereditariedade/genética , Homozigoto , Humanos , Nefropatias/genética , Masculino , Proteínas de Membrana/genética , Camundongos , Fenótipo , RNA Longo não Codificante/genética , Sistema Urinário/anormalidades , Anormalidades Urogenitais/genética , Peixe-Zebra
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