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1.
BMC Neurol ; 20(1): 314, 2020 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-32847546

RESUMO

BACKGROUND: Kartagener syndrome is an autosomal recessive inherited disorder of primary ciliary dyskinesia. Moyamoya syndrome refers to a moyamoya angiopathy associated with other neurological and/or extra-neurological symptoms, or due to a well identified acquired or inherited cause. We herein reported a case of a 48-year-old woman who was favored the diagnosis of Kartagener syndrome and moyamoya syndrome. The whole genome sequencing and bioinformatics analysis showed a homozygotic nonsense mutation in the dynein, axonemal, heavy chain (DNAH) 5 gene, and heterozygotic missense mutation in the DNAH11 gene. This is the first report of the co-occurrence of the two rare diseases. CASE PRESENTATION: A case of a 48-year-old woman was presented with hemiplegia and slurred speech. The magnetic resonance imaging of the brain confirmed acute cerebral infarction in the right basal ganglia region, semi-oval center, insular lobe, and frontal parietal lobe. The electrocardiogram showed inverted "P" waves in L1 and AVL on left-sided chest leads and computed tomography scan of the chest showed bronchiectasis changes, cardiac shadow and apex on the right side, and situs inversus of aortic arch position. The digital subtraction angiography showed inversion of the aortic arch, and bilateral internal carotid arteries are occluded from the ophthalmic segment. The clinical, radiological, and laboratory findings made the diagnosis of Kartagener syndrome and moyamoya syndrome. The whole genome sequencing and bioinformatics analysis showed a homozygotic nonsense mutation in DNAH5 gene, and heterozygotic missense mutation in the DNAH11 gene. CONCLUSION: The combined mutation of DNAH5 and DNAH11 may lead to the overlapping dysfunction of motile and nonmotile cilia, which contribute to the co-occurrence of Kartagener syndrome and moyamoya syndrome. Our report deserves further confirm by more case reports.


Assuntos
Dineínas do Axonema/genética , Síndrome de Kartagener/diagnóstico , Doença de Moyamoya/diagnóstico , Encéfalo/patologia , Feminino , Humanos , Síndrome de Kartagener/genética , Síndrome de Kartagener/patologia , Pessoa de Meia-Idade , Mutação , Mutação de Sentido Incorreto , Sequenciamento Completo do Genoma
2.
Cells ; 8(8)2019 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-31443223

RESUMO

Primary ciliary dyskinesia (PCD) is a rare autosomal recessive disorder characterized by dysfunction of motile cilia causing ineffective mucus clearance and organ laterality defects. In this study, two unrelated Portuguese children with strong PCD suspicion underwent extensive clinical and genetic assessments by whole-exome sequencing (WES), as well as ultrastructural analysis of cilia by transmission electron microscopy (TEM) to identify their genetic etiology. These analyses confirmed the diagnostic of Kartagener syndrome (KS) (PCD with situs inversus). Patient-1 showed a predominance of the absence of the inner dynein arms with two disease-causing variants in the CCDC40 gene. Patient-2 showed the absence of both dynein arms and WES disclosed two novel high impact variants in the DNAH5 gene and two missense variants in the DNAH7 gene, all possibly deleterious. Moreover, in Patient-2, functional data revealed a reduction of gene expression and protein mislocalization in both genes' products. Our work calls the researcher's attention to the complexity of the PCD and to the possibility of gene interactions modelling the PCD phenotype. Further, it is demonstrated that even for well-known PCD genes, novel pathogenic variants could have importance for a PCD/KS diagnosis, reinforcing the difficulty of providing genetic counselling and prenatal diagnosis to families.


Assuntos
Cílios/ultraestrutura , Síndrome de Kartagener/genética , Síndrome de Kartagener/patologia , Dineínas do Axonema/genética , Criança , Dineínas/genética , Feminino , Testes Genéticos , Humanos , Masculino , Mutação , Portugal , Proteínas/genética , Sequenciamento Completo do Exoma/métodos
3.
BMC Pulm Med ; 19(1): 135, 2019 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-31345208

RESUMO

BACKGROUND: Primary ciliary dyskinesia (PCD) is a rare genetic disorder. Although the genetic tests and new diagnostic algorithms have recently been recommended, clinical signs and electron microscope (EM) findings have historically been the mainstays of diagnosis in Asia. To characterize PCD previously reported in Japan, we conducted a systematic review and meta-analysis. METHODS: A search using MEDLINE, EMBASE, and Japana Centra Revuo Medicina (in Japanese) databases was carried out to identify articles reporting PCD, Kartagener syndrome, or immotile cilia syndrome in Japanese patients and published between 1985 and 2015. RESULTS: After excluding duplication from 334 reports, we extracted 316 patients according to the criteria. Diagnosis was most frequently made in adulthood (148 patients [46.8%] ≥ 18 years old, 24 patients [7.6%] < 1 year old, 68 patients [21.5%] 1-17 years old and 76 patients [24.1%] lacking information). Of the 230 patients (72.8%) who received EM examination, there were patients with inner dynein arm (IDA) defects (n = 55; 23.9%), outer dynein arm (ODA) defects (14; 6.1%), both ODA and IDA defects (57; 24.8%), other structural abnormalities (25; 10.9%), no abnormalities (4; 1.7%), and no detailed conclusion or description (75; 32.6%). CONCLUSION: Delayed diagnosis of this congenital disease with high frequency of IDA defects and low frequency of ODA defects appear to be historical features of PCD reported in Japan, when EM was a main diagnostic tool. This review highlights problems experienced in this field, and provides basic information to establish a modernized PCD diagnosis and management system in the future.


Assuntos
Dineínas/deficiência , Síndrome de Kartagener/diagnóstico , Cílios/fisiologia , Cílios/ultraestrutura , Diagnóstico Tardio , Dineínas/ultraestrutura , Humanos , Japão , Síndrome de Kartagener/patologia , Microscopia Eletrônica
4.
J Assist Reprod Genet ; 36(8): 1683-1700, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31273583

RESUMO

PROPOSE: To study CCDC103 expression profiles and understand how pathogenic variants in CCDC103 affect its expression profile at mRNA and protein level. METHODS: To increase the knowledge about the CCDC103, we attempted genotype-phenotype correlations in two patients carrying novel homozygous (missense and frameshift) CCDC103 variants. Whole-exome sequencing, quantitative PCR, Western blot, electron microscopy, immunohistochemistry, immunocytochemistry, and immunogold labelling were performed to characterize CCDC103 expression profiles in reproductive and somatic cells. RESULTS: Our data demonstrate that pathogenic variants in CCDC103 gene negatively affect gene and protein expression in both patients who presented absence of DA on their axonemes. Further, we firstly report that CCDC103 is expressed at different levels in reproductive tissues and somatic cells and described that CCDC103 protein forms oligomers with tissue-specific sizes, which suggests that CCDC103 possibly undergoes post-translational modifications. Moreover, we reported that CCDC103 was restricted to the midpiece of sperm and is present at the cytoplasm of the other cells. CONCLUSIONS: Overall, our data support the CCDC103 involvement in PCD and suggest that CCDC103 may have different assemblies and roles in cilia and sperm flagella biology that are still unexplored.


Assuntos
Axonema/patologia , Transtornos da Motilidade Ciliar/genética , Infertilidade Masculina/patologia , Síndrome de Kartagener/genética , Proteínas Associadas aos Microtúbulos/genética , Mutação , Cauda do Espermatozoide/patologia , Axonema/genética , Transtornos da Motilidade Ciliar/patologia , Dineínas/metabolismo , Feminino , Humanos , Infertilidade Masculina/etiologia , Síndrome de Kartagener/patologia , Masculino , Pessoa de Meia-Idade , Reprodução , Situs Inversus/genética , Situs Inversus/patologia , Cauda do Espermatozoide/metabolismo
5.
Int J Pediatr Otorhinolaryngol ; 115: 94-96, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30368402

RESUMO

OBJECTIVE: To determine whether adenoid epithelium is superior to nasal mucosa for biopsy of ciliated epithelium for electron microscopy (EM) to evaluate pediatric patients with rhinosinusitis for primary ciliary dyskinesia (PCD). METHODS: A retrospective review compared electron microscopic results in children with chronic or recurrent rhinosinusitis who underwent both adenoidectomy or nasopharyngeal biopsy and nasal mucosa biopsy in the course of evaluation for PCD at a tertiary care institution. RESULTS: Forty pediatric patients met inclusion criteria for this study. Nine of these patients had a prior adenoidectomy and therefore underwent nasopharyngeal biopsy for collection of adenoid tissue. All nine of the nasopharyngeal biopsies and 25 of the 31 (80.6%) adenoid biopsies had sufficient cilia for EM evaluation of the ultrastructure. Of the 40 patients who also had a nasal biopsy, only 12 (30.0%) had sufficient cilia for EM analysis. The distribution of sufficient versus insufficient cilia for analysis between adenoid and nasal mucosa was statistically significant (P < 0.05). Abnormal cilia were found in only 2.5% of our patients. CONCLUSIONS: In current practice, the nasal cavity is a common location for obtaining ciliated epithelium for EM analysis, as it is easily accessible for biopsy and the procedure itself causes relatively low patient morbidity. Chronic rhinosinusitis, however, has been associated with decreased cilia density on nasal respiratory epithelium. Given that adenoidectomies are often performed in children with chronic rhinosinusitis, our data suggest that adenoid tissue is a better source of ciliated tissue for analysis compared to turbinate epithelium.


Assuntos
Tonsila Faríngea/patologia , Síndrome de Kartagener/patologia , Mucosa Nasal/patologia , Rinite/patologia , Sinusite/patologia , Adenoidectomia , Biópsia/métodos , Criança , Pré-Escolar , Doença Crônica , Cílios/patologia , Humanos , Lactente , Microscopia Eletrônica , Recidiva , Estudos Retrospectivos
6.
Respir Res ; 19(1): 125, 2018 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-29940967

RESUMO

BACKGROUND: Primary ciliary dyskinesia can result from a number of different ciliary defects that adversely affect ciliary function resulting markedly reduced or absent mucociliary clearance. Improvement in diagnostic testing is an area of current research. During diagnostic evaluation of PCD we observed ciliated conical protrusions from part of the apical surface of ciliated cells in those diagnosed with PCD. The aim of this study was to investigate if this abnormality was specific to PCD. METHODS: Epithelial edges from 67 consecutively diagnosed PCD patients, 67 patients consecutively referred for PCD diagnostic testing in whom PCD was excluded, 22 with asthma and 18 with Cystic Fibrosis (CF) were studied retrospectively in a blinded manner using light microscopy. RESULTS: Forty six out of 67 patients with PCD had ciliated conical epithelial protrusions, whereas none were seen in patients where PCD was excluded, or in patients with asthma or CF. The sensitivity, specificity, positive predictive value and negative predictive value for the presence of the ciliated conical protrusions to predict a diagnosis of PCD were 76.5, 100, 100 and 77% respectively. CONCLUSIONS: Characteristic ciliated conical protrusions from ciliated epithelial cells maybe a useful pointer to the diagnosis of PCD. However, their absence does not exclude the diagnosis of PCD.


Assuntos
Cílios/patologia , Cílios/fisiologia , Síndrome de Kartagener/patologia , Depuração Mucociliar/fisiologia , Mucosa Respiratória/patologia , Mucosa Respiratória/fisiologia , Células Cultivadas , Humanos
7.
Am J Physiol Lung Cell Mol Physiol ; 314(6): L909-L921, 2018 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-29493257

RESUMO

The respiratory tract is lined with multiciliated epithelial cells that function to move mucus and trapped particles via the mucociliary transport apparatus. Genetic and acquired ciliopathies result in diminished mucociliary clearance, contributing to disease pathogenesis. Recent innovations in imaging technology have advanced our understanding of ciliary motion in health and disease states. Application of imaging modalities including transmission electron microscopy, high-speed video microscopy, and micron-optical coherence tomography could improve diagnostics and be applied for precision medicine. In this review, we provide an overview of ciliary motion, imaging modalities, and ciliopathic diseases of the respiratory system including primary ciliary dyskinesia, cystic fibrosis, chronic obstructive pulmonary disease, and idiopathic pulmonary fibrosis.


Assuntos
Cílios , Fibrose Cística , Síndrome de Kartagener , Depuração Mucociliar/genética , Doença Pulmonar Obstrutiva Crônica , Animais , Cílios/genética , Cílios/metabolismo , Cílios/patologia , Fibrose Cística/genética , Fibrose Cística/metabolismo , Fibrose Cística/patologia , Humanos , Síndrome de Kartagener/genética , Síndrome de Kartagener/metabolismo , Síndrome de Kartagener/patologia , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia
8.
Biomed Res Int ; 2018: 1854269, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29511670

RESUMO

Primary ciliary dyskinesia (PCD) is a clinical rare peculiar disorder, mainly featured by respiratory infection, tympanitis, nasosinusitis, and male infertility. Previous study demonstrated it is an autosomal recessive disease and by 2017 almost 40 pathologic genes have been identified. Among them are the leucine-rich repeat- (LRR-) containing 6 (LRRC6) codes for a 463-amino-acid cytoplasmic protein, expressed distinctively in motile cilia cells, including the testis cells and the respiratory epithelial cells. In this study, we applied whole-exome sequencing combined with PCD-known genes filtering to explore the genetic lesion of a PCD patient. A novel compound heterozygous mutation in LRRC6 (c.183T>G/p.N61K; c.179-1G>A) was identified and coseparated in this family. The missense mutation (c.183T>G/p.N61K) may lead to a substitution of asparagine by lysine at position 61 in exon 3 of LRRC6. The splice site mutation (c.179-1G>A) may cause a premature stop codon in exon 4 and decrease the mRNA levels of LRRC6. Both mutations were not present in our 200 local controls, dbSNP, and 1000 genomes. Three bioinformatics programs also predicted that both mutations are deleterious. Our study not only further supported the importance of LRRC6 in PCD, but also expanded the spectrum of LRRC6 mutations and will contribute to the genetic diagnosis and counseling of PCD patients.


Assuntos
Cílios/genética , Síndrome de Kartagener/genética , Proteínas/genética , Sequenciamento Completo do Exoma , China , Cílios/patologia , Proteínas do Citoesqueleto , Exoma/genética , Feminino , Predisposição Genética para Doença , Heterozigoto , Humanos , Síndrome de Kartagener/diagnóstico por imagem , Síndrome de Kartagener/patologia , Masculino , Mutação , Linhagem , Tomografia Computadorizada por Raios X
9.
Reprod Biol Endocrinol ; 16(1): 10, 2018 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-29402277

RESUMO

Primary ciliary dyskinesia (PCD) is a rare, autosomal recessive disease with abnormalities in the structure of cilia, causing impairment of muco-ciliary clearance with respiratory tract infections, heterotaxia and abnormal sperm motility with male infertility. Here, with a comprehensive literature review, we report a couple with an infertility history of 9 years and three unsuccessful IVF treatments, where male partner has Kartagener's Syndrome, a subtype of PCD, displaying recurrent respiratory infections, dextrocardia and total asthenozoospermia. His diagnosis was verified with transmission electron microscopy and genetic mutation screening, revealing total absence of dynein arms in sperm tails and homozygous mutation in the ZMYND10, heterozygous mutations in the ARMC4 and DNAH5 genes. Laser assisted viability assay (LAVA) was performed by shooting the sperm tails during sperm retrieval for microinjection, following detection of pentoxifylline resistant immotile sperm. Live births of healthy triplets, one boy and two monozygotic girls, was achieved after double blastocyst transfer.


Assuntos
Infertilidade Masculina/terapia , Síndrome de Kartagener/complicações , Lasers , Nascimento Vivo , Análise do Sêmen/métodos , Espermatozoides/fisiologia , Sobrevivência Celular , Feminino , Humanos , Infertilidade Masculina/etiologia , Infertilidade Masculina/genética , Síndrome de Kartagener/genética , Síndrome de Kartagener/patologia , Masculino , Pentoxifilina , Gravidez , Injeções de Esperma Intracitoplásmicas , Espermatozoides/ultraestrutura
12.
Paediatr Respir Rev ; 24: 19-20, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28687245

RESUMO

Primary ciliary dyskinesia is an inherited disease characterized by impaired ciliary function leading to diverse clinical manifestations, including chronic sinopulmonary disease, persistent middle ear effusions, laterality defects, and infertility. Our understanding of the complex genetics and functional phenotypes of primary ciliary dyskinesia has rapidly grown, and over 35 disease-associated genes have been identified, which segregate into genes that encode axonemal motor proteins, regulatory proteins within the cilium, and cytoplasmic proteins involved in ciliary assembly. These findings have yielded unexpected insights into the clinical heterogeneity of disease and are beginning to revolutionize diagnostic testing for primary ciliary dyskinesia.


Assuntos
Cílios/ultraestrutura , Testes Genéticos , Síndrome de Kartagener/diagnóstico , Cílios/genética , Genótipo , Humanos , Síndrome de Kartagener/genética , Síndrome de Kartagener/patologia , Microscopia Eletrônica de Transmissão , Mutação , Fenótipo
13.
Ann Am Thorac Soc ; 14(7): 1184-1196, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28481653

RESUMO

RATIONALE: Primary ciliary dyskinesia (PCD) is a rare disorder causing chronic otosinopulmonary disease, generally diagnosed through evaluation of respiratory ciliary ultrastructure and/or genetic testing. Nasal nitric oxide (nNO) measurement is used as a PCD screening test because patients with PCD have low nNO levels, but its value as a diagnostic test remains unknown. OBJECTIVES: To perform a systematic review to assess the utility of nNO measurement (index test) as a diagnostic tool compared with the reference standard of electron microscopy (EM) evaluation of ciliary defects and/or detection of biallelic mutations in PCD genes. DATA SOURCES: Ten databases were searched for reference sources from database inception through July 29, 2016. DATA EXTRACTION: Study inclusion was limited to publications with rigorous nNO index testing, reference standard diagnostic testing with EM and/or genetics, and calculable diagnostic accuracy information for cooperative patients (generally >5 yr old) with high suspicion of PCD. SYNTHESIS: Meta-analysis provided a summary estimate for sensitivity and specificity and a hierarchical summary receiver operating characteristic curve. The Quality Assessment of Diagnostic Accuracy Studies-2 tool was used to assess study quality, and Grading of Recommendations Assessment, Development, and Evaluation was used to assess the certainty of evidence. In 12 study populations (1,344 patients comprising 514 with PCD and 830 without PCD), using a reference standard of EM alone or EM and/or genetic testing, summary sensitivity was 97.6% (92.7-99.2) and specificity was 96.0% (87.9-98.7), with a positive likelihood ratio of 24.3 (7.6-76.9), a negative likelihood ratio of 0.03 (0.01-0.08), and a diagnostic odds ratio of 956.8 (141.2-6481.5) for nNO measurements. After studies using EM alone as the reference standard were excluded, the seven studies using an extended reference standard of EM and/or genetic testing showed a summary sensitivity of nNO measurements of 96.3% (88.7-98.9) and specificity of 96.4% (85.1-99.2), with a positive likelihood ratio of 26.5 (5.9-119.1), a negative likelihood ratio of 0.04 (0.01-0.12), and a diagnostic odds ratio of 699.3 (67.4-7256.0). Certainty of the evidence was graded as moderate. CONCLUSIONS: nNO is a sensitive and specific test for PCD in cooperative patients (generally >5 yr old) with high clinical suspicion for this disease. With a moderate level of evidence, this meta-analysis confirms that nNO testing using velum closure maneuvers has diagnostic accuracy similar to EM and/or genetic testing for PCD when cystic fibrosis is ruled out. Thus, low nNO values accompanied by an appropriate clinical phenotype could be used as a diagnostic PCD test, though EM and/or genetics will continue to provide confirmatory information.


Assuntos
Síndrome de Kartagener/diagnóstico , Óxido Nítrico/análise , Humanos , Síndrome de Kartagener/patologia , Microscopia Eletrônica , Sensibilidade e Especificidade
14.
Vet Pathol ; 54(5): 802-812, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28494707

RESUMO

Mucociliary clearance is a main defense mechanism of the respiratory tract, which can be inherently impaired in primary ciliary dyskinesia (PCD) or reversibly altered in secondary ciliary dyskinesia (SCD). Limited diagnostic test availability likely leads to misdiagnosis or underdiagnosis of PCD in animals. This study evaluated the light and transmission electron microscopy (TEM) changes in the respiratory mucosa of 15 dogs with chronic respiratory disease suspected of PCD. Necropsy was performed in 1 case and 2 dogs were used as negative controls. PCD was confirmed in 1 dog, which presented with chronic otitis, bronchopneumonia, hydrocephalus and ultrastructural abnormalities in 84% of the assessed cilia, including absence of dynein arms and microtubular changes. The 14 other cases showed only nonspecific alterations, such as ciliary disorientation, compound cilia, ciliary edema, and axoneme bubbles in a minority of the evaluated cilia and were classified as SCD. Ciliary ultrastructural analysis can confirm a diagnosis of PCD if specific abnormalities exist. TEM remains an important investigational tool in veterinary medicine, as no other specific test for PCD in dogs has been standardized yet.


Assuntos
Doenças do Cão/diagnóstico , Síndrome de Kartagener/veterinária , Animais , Doença Crônica/veterinária , Cílios/patologia , Cílios/ultraestrutura , Doenças do Cão/patologia , Cães , Feminino , Síndrome de Kartagener/diagnóstico , Síndrome de Kartagener/patologia , Masculino , Microscopia Eletrônica de Transmissão/veterinária , Depuração Mucociliar , Mucosa Respiratória/patologia , Mucosa Respiratória/ultraestrutura
15.
Ital J Pediatr ; 43(1): 34, 2017 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-28403885

RESUMO

BACKGROUND: Primary ciliary dyskinesia (PCD) and cystic fibrosis (CF) are increasingly compared. There are no chest magnetic resonance imaging (MRI) comparative studies of PCD and CF. We assessed clinical, functional, microbiological and MRI findings in PCD and mild CF patients in order to evaluate different expression of lung disease. METHODS: Twenty PCD (15.1 years) and 20 CF subjects with mild respiratory impairment (16 years, 70% with pancreatic insufficiency) underwent MRI, spirometry, and sputum cultures when clinically stable. MRI was scored using the modified Helbich system. RESULTS: PCD was diagnosed later than CF (9.9 versus 0.6 years, p = 0.03), despite earlier symptoms (0.1 versus 0.6 years, p = 0.02). In the year preceding the study, patients from both groups underwent two systemic antibiotic courses (p = 0.48). MRI total scores were 11.6 ± 0.7 and 9.1 ± 1 in PCD and CF, respectively. FEV1 and FVC Z-scores were -1.75 (range, -4.6-0.7) and -0.6 (-3.9-1.8) in PCD, and -0.9 (range, -5.4-2.3) and -0.3 (-3.4-2.5) in CF, respectively. No difference was found between lung function or structure, despite a higher MRI subscore of collapse/consolidation in PCD versus CF (1.6 ± 0.1 and 0.6 ± 0.2, p < 0.001). These findings were confirmed after data-control for diagnostic delay. Pseudomonas aeruginosa and Staphylococcus aureus were more frequent in CF than in PCD (p = 0.05 and p = 0.003, respectively). CONCLUSIONS: MRI is a valuable radiation-free tool for comparative PCD and CF lung disease assessment. Patients with PCD may exhibit similar MRI and lung function changes as CF subjects with mild pulmonary disease. Delay in PCD diagnosis is unlikely the only determinant of similarities.


Assuntos
Fibrose Cística/diagnóstico , Síndrome de Kartagener/diagnóstico , Imagem por Ressonância Magnética/métodos , Espirometria/métodos , Tomografia Computadorizada por Raios X/métodos , Adolescente , Adulto , Criança , Estudos de Coortes , Fibrose Cística/patologia , Diagnóstico Diferencial , Feminino , Humanos , Síndrome de Kartagener/patologia , Masculino , Projetos Piloto , Estudos Prospectivos , Índice de Gravidade de Doença , Escarro/microbiologia , Adulto Jovem
16.
Nat Commun ; 8: 14279, 2017 02 08.
Artigo em Inglês | MEDLINE | ID: mdl-28176794

RESUMO

By moving essential body fluids and molecules, motile cilia and flagella govern respiratory mucociliary clearance, laterality determination and the transport of gametes and cerebrospinal fluid. Primary ciliary dyskinesia (PCD) is an autosomal recessive disorder frequently caused by non-assembly of dynein arm motors into cilia and flagella axonemes. Before their import into cilia and flagella, multi-subunit axonemal dynein arms are thought to be stabilized and pre-assembled in the cytoplasm through a DNAAF2-DNAAF4-HSP90 complex akin to the HSP90 co-chaperone R2TP complex. Here, we demonstrate that large genomic deletions as well as point mutations involving PIH1D3 are responsible for an X-linked form of PCD causing disruption of early axonemal dynein assembly. We propose that PIH1D3, a protein that emerges as a new player of the cytoplasmic pre-assembly pathway, is part of a complementary conserved R2TP-like HSP90 co-chaperone complex, the loss of which affects assembly of a subset of inner arm dyneins.


Assuntos
Proteínas Reguladoras de Apoptose/genética , Dineínas do Axonema/metabolismo , Genes Ligados ao Cromossomo X/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Síndrome de Kartagener/genética , Proteínas dos Microtúbulos/genética , Chaperonas Moleculares/genética , Adolescente , Adulto , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Axonema/patologia , Criança , Pré-Escolar , Cílios/patologia , Cílios/ultraestrutura , Citoplasma/patologia , Modelos Animais de Doenças , Feminino , Doenças Genéticas Ligadas ao Cromossomo X/patologia , Células HEK293 , Proteínas de Choque Térmico HSP90/metabolismo , Humanos , Recém-Nascido , Peptídeos e Proteínas de Sinalização Intracelular , Síndrome de Kartagener/patologia , Masculino , Microscopia Eletrônica de Transmissão , Linhagem , Filogenia , Mutação Puntual , Dobramento de Proteína , Alinhamento de Sequência , Deleção de Sequência , Motilidade Espermática/genética , Sequenciamento Completo do Exoma , Peixe-Zebra
17.
Eur Spine J ; 26(6): 1595-1599, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28180983

RESUMO

PURPOSE: Primary ciliary dyskinesia (PCD) is a respiratory syndrome in which 'random' organ orientation can occur; with approximately 46% of patients developing situs inversus totalis at organogenesis. The aim of this study was to explore the relationship between organ anatomy and curve convexity by studying the prevalence and convexity of idiopathic scoliosis in PCD patients with and without situs inversus. METHODS: Chest radiographs of PCD patients were systematically screened for existence of significant lateral spinal deviation using the Cobb angle. Positive values represented right-sided convexity. Curve convexity and Cobb angles were compared between PCD patients with situs inversus and normal anatomy. RESULTS: A total of 198 PCD patients were screened. The prevalence of scoliosis (Cobb >10°) and significant spinal asymmetry (Cobb 5-10°) was 8 and 23%, respectively. Curve convexity and Cobb angle were significantly different within both groups between situs inversus patients and patients with normal anatomy (P ≤ 0.009). Moreover, curve convexity correlated significantly with organ orientation (P < 0.001; ϕ = 0.882): In 16 PCD patients with scoliosis (8 situs inversus and 8 normal anatomy), except for one case, matching of curve convexity and orientation of organ anatomy was observed: convexity of the curve was opposite to organ orientation. CONCLUSIONS: This study supports our hypothesis on the correlation between organ anatomy and curve convexity in scoliosis: the convexity of the thoracic curve is predominantly to the right in PCD patients that were 'randomized' to normal organ anatomy and to the left in patients with situs inversus totalis.


Assuntos
Síndrome de Kartagener/patologia , Escoliose/patologia , Situs Inversus/patologia , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Síndrome de Kartagener/complicações , Síndrome de Kartagener/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Prevalência , Radiografia , Escoliose/diagnóstico por imagem , Escoliose/epidemiologia , Escoliose/etiologia , Situs Inversus/complicações , Situs Inversus/diagnóstico por imagem , Adulto Jovem
18.
J Assist Reprod Genet ; 34(2): 275-281, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27988889

RESUMO

PURPOSE: Kartagener syndrome (KS), also known as visceral inversion-nasosinusitis-bronchiectasis syndrome, or familial bronchiectasis, is an autosomal recessive inherited disease. In this study, through two cases of KS, we aimed to assess the clinical and genetic characteristics of KS caused by DNAH5 mutations. METHODS: The two cases of KS from the same family underwent extensive clinical assessments, with next-generation DNA sequencing and bioinformatics analysis to identify pathogenic genes. In addition, Sanger sequencing was used to verify the pedigrees. RESULTS: The present study employed a directional capture strategy for hereditary disease screening, which correctly identified the virulence sites in the pedigree, and facilitated the differential diagnosis among multiple genes. Two novel mutations were detected in DNAH5: c.7778C>T (missense mutation) and c.13729G>A (nonsense mutation). They were not found in dbSNP, 1000 Genomes, and ExAC. CONCLUSIONS: These findings demonstrated that new DNAH5 mutations could be used for molecular diagnosis of KS, providing families with genetic counseling and prenatal diagnosis.


Assuntos
Dineínas do Axonema/genética , Bronquiectasia/genética , Síndrome de Kartagener/genética , Bronquiectasia/diagnóstico , Bronquiectasia/patologia , Criança , Códon sem Sentido/genética , Códon sem Sentido/isolamento & purificação , Feminino , Testes Genéticos , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Síndrome de Kartagener/diagnóstico , Síndrome de Kartagener/patologia , Masculino , Mutação de Sentido Incorreto/genética , Linhagem , Fenótipo
19.
Am J Physiol Lung Cell Mol Physiol ; 312(2): L258-L267, 2017 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-27979861

RESUMO

Air-liquid interface (ALI) culture of primary airway epithelial cells enables mucociliary differentiation providing an in vitro model of the human airway, but their proliferative potential is limited. To extend proliferation, these cells were previously transduced with viral oncogenes or mouse Bmi-1 + hTERT, but the resultant cell lines did not undergo mucociliary differentiation. We hypothesized that use of human BMI-1 alone would increase the proliferative potential of bronchial epithelial cells while retaining their mucociliary differentiation potential. Cystic fibrosis (CF) and non-CF bronchial epithelial cells were transduced by lentivirus with BMI-1 and then their morphology, replication kinetics, and karyotype were assessed. When differentiated at ALI, mucin production, ciliary function, and transepithelial electrophysiology were measured. Finally, shRNA knockdown of DNAH5 in BMI-1 cells was used to model primary ciliary dyskinesia (PCD). BMI-1-transduced basal cells showed normal cell morphology, karyotype, and doubling times despite extensive passaging. The cell lines underwent mucociliary differentiation when cultured at ALI with abundant ciliation and production of the gel-forming mucins MUC5AC and MUC5B evident. Cilia displayed a normal beat frequency and 9+2 ultrastructure. Electrophysiological characteristics of BMI-1-transduced cells were similar to those of untransduced cells. shRNA knockdown of DNAH5 in BMI-1 cells produced immotile cilia and absence of DNAH5 in the ciliary axoneme as seen in cells from patients with PCD. BMI-1 delayed senescence in bronchial epithelial cells, increasing their proliferative potential but maintaining mucociliary differentiation at ALI. We have shown these cells are amenable to genetic manipulation and can be used to produce novel disease models for research and dissemination.


Assuntos
Brônquios/citologia , Diferenciação Celular , Cílios/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Muco/metabolismo , Complexo Repressor Polycomb 1/metabolismo , Animais , Dineínas do Axonema/metabolismo , Proliferação de Células , Forma Celular , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Dineínas/metabolismo , Impedância Elétrica , Fenômenos Eletrofisiológicos , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , Síndrome de Kartagener/metabolismo , Síndrome de Kartagener/patologia , Síndrome de Kartagener/fisiopatologia , Cariotipagem , Camundongos , Microtúbulos/metabolismo , Modelos Biológicos , Fenótipo , Transdução Genética
20.
BMC Microbiol ; 16(1): 200, 2016 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-27586172

RESUMO

BACKGROUND: Burkholderia cenocepacia is a Gram-negative, opportunistic pathogen that is a cause of morbidity and mortality in patients with cystic fibrosis (CF). Research efforts over the past few decades contributed to our understanding of these infections by identifying virulence factors. However, little is known about how this pathogen adapts to the harsh environment found inside the CF airways, which is characterized by a unique mucus containing high concentrations of inflammatory markers. The current study developed a novel model to further investigate this phenomenon. RESULTS: Monolayers of human A549 lung carcinoma cells (HLCCs) were exposed to a mixture of artificial CF sputum medium (ASMDM) in tissue culture growth medium, and subsequently infected with B. cenocepacia K56-2 for 24 h. The data showed that this model supported B. cenocepacia growth. In addition, consistent with similar studies using current models such as CF airway tissue samples, HLCC viability was reduced by more than 70 % when grown in 60 % ASMDM and infected with B. cenocepacia compared to mock-infected controls and medium alone. Furthermore, the amount of B. cenocepacia cells associated with the HLCC monolayer was more than 10 times greater in 60 % ASMDM when compared to medium controls. CONCLUSIONS: These findings suggest that HLCC monolayers in 60 % ASMDM serve as a valid alternative to study B. cenocepacia infections in patients with CF, and possibly other chronic diseases of the airways. Furthermore, the results obtained in this study suggest an important role for CF sputum in B. cenocepacia pathogenesis.


Assuntos
Infecções por Burkholderia/microbiologia , Burkholderia cenocepacia/patogenicidade , Fibrose Cística/microbiologia , Síndrome de Kartagener/microbiologia , Neoplasias Pulmonares/microbiologia , Células A549 , Infecções por Burkholderia/patologia , Burkholderia cenocepacia/efeitos dos fármacos , Doença Crônica , Meios de Cultivo Condicionados , Humanos , Síndrome de Kartagener/patologia , Neoplasias Pulmonares/patologia , Viabilidade Microbiana , Escarro/microbiologia , Tetraciclina/farmacologia , Fatores de Virulência
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