scholarly journals Primary Ciliary Dyskinesia in Mice Lacking the Novel Ciliary Protein Pcdp1

2007 ◽  
Vol 28 (3) ◽  
pp. 949-957 ◽  
Author(s):  
Lance Lee ◽  
Dean R. Campagna ◽  
Jack L. Pinkus ◽  
Howard Mulhern ◽  
Todd A. Wyatt ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Male Infertility ◽  
Primary Ciliary Dyskinesia ◽  
Single Gene ◽  
Ependymal Cells ◽  
The Novel ◽  
Gene Encoding ◽  
Respiratory Epithelial ◽  
Ciliated Epithelial Cells ◽  
Ciliary Dyskinesia

ABSTRACT Primary ciliary dyskinesia (PCD) results from ciliary dysfunction and is commonly characterized by sinusitis, male infertility, hydrocephalus, and situs inversus. Mice homozygous for the nm1054 mutation develop phenotypes associated with PCD. On certain genetic backgrounds, homozygous mutants die perinatally from severe hydrocephalus, while mice on other backgrounds have an accumulation of mucus in the sinus cavity and male infertility. Mutant sperm lack mature flagella, while respiratory epithelial cilia are present but beat at a slower frequency than wild-type cilia. Transgenic rescue demonstrates that the PCD in nm1054 mutants results from the loss of a single gene encoding the novel primary ciliary dyskinesia protein 1 (Pcdp1). The Pcdp1 gene is expressed in spermatogenic cells and motile ciliated epithelial cells. Immunohistochemistry shows that Pcdp1 protein localizes to sperm flagella and the cilia of respiratory epithelial cells and brain ependymal cells in both mice and humans. This study demonstrates that Pcdp1 plays an important role in ciliary and flagellar biogenesis and motility, making the nm1054 mutant a useful model for studying the molecular genetics and pathogenesis of PCD.

Bi-allelic BRWD1 variants cause male infertility with asthenoteratozoospermia and likely primary ciliary dyskinesia

2021 ◽  
Author(s):  
Ting Guo ◽  
Chao-Feng Tu ◽  
Dan-Hui Yang ◽  
Shui-Zi Ding ◽  
Cheng Lei ◽  
...  
Keyword(s):  
Male Infertility ◽  
Primary Ciliary Dyskinesia ◽  
Ciliary Dyskinesia

scholarly journals A Revised Protocol for Culture of Airway Epithelial Cells as a Diagnostic Tool for Primary Ciliary Dyskinesia

2020 ◽  
Vol 9 (11) ◽  
pp. 3753
Author(s):  
Janice L. Coles ◽  
James Thompson ◽  
Katie L. Horton ◽  
Robert A. Hirst ◽  
Paul Griffin ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Primary Ciliary Dyskinesia ◽  
High Speed ◽  
Ex Vivo ◽  
Cell Damage ◽  
Airway Epithelial Cells ◽  
University Hospital ◽  
Mitigation Measures ◽  
Diagnostic Certainty ◽  
Ciliary Dyskinesia

Air–liquid interface (ALI) culture of nasal epithelial cells is a valuable tool in the diagnosis and research of primary ciliary dyskinesia (PCD). Ex vivo samples often display secondary dyskinesia from cell damage during sampling, infection or inflammation confounding PCD diagnostic results. ALI culture enables regeneration of healthy cilia facilitating differentiation of primary from secondary ciliary dyskinesia. We describe a revised ALI culture method adopted from April 2018 across three collaborating PCD diagnostic sites, including current University Hospital Southampton COVID-19 risk mitigation measures, and present results. Two hundred and forty nasal epithelial cell samples were seeded for ALI culture and 199 (82.9%) were ciliated. Fifty-four of 83 (63.9%) ex vivo samples which were originally equivocal or insufficient provided diagnostic information following in vitro culture. Surplus basal epithelial cells from 181 nasal brushing samples were frozen in liquid nitrogen; 39 samples were ALI-cultured after cryostorage and all ciliated. The ciliary beat patterns of ex vivo samples (by high-speed video microscopy) were recapitulated, scanning electron microscopy demonstrated excellent ciliation, and cilia could be immuno-fluorescently labelled (anti-alpha-tubulin and anti-RSPH4a) in representative cases that were ALI-cultured after cryostorage. In summary, our ALI culture protocol provides high ciliation rates across three centres, minimising patient recall for repeat brushing biopsies and improving diagnostic certainty. Cryostorage of surplus diagnostic samples was successful, facilitating PCD research.

scholarly journals Mutations in C11ORF70 cause primary ciliary dyskinesia with randomization of left/right body asymmetry due to outer and inner dynein arm defects

2017 ◽  
Author(s):  
Inga M. Höben ◽  
Rim Hjeij ◽  
Heike Olbrich ◽  
Gerard W. Dougherty ◽  
Tabea Menchen ◽  
...  
Keyword(s):  
Male Infertility ◽  
Primary Ciliary Dyskinesia ◽  
Genetic Defect ◽  
Clinical Importance ◽  
Airway Disease ◽  
Cytoplasmic Dynein ◽  
Loss Of Function ◽  
Reading Frame ◽  
Dynein Arms ◽  
Ciliary Dyskinesia

AbstractPrimary ciliary dyskinesia (PCD) is characterized by chronic airway disease, male infertility and randomization of the left/right body axis caused by defects of motile cilia and sperm flagella. We identified loss-of-function mutations in the open reading frame C11ORF70 in PCD individuals from five distinct families. Transmission electron microscopy analyses and high resolution immunofluorescence microscopy demonstrate that loss-of-function mutations in C11ORF70 cause immotility of respiratory cilia and sperm flagella, respectively, due to loss of axonemal outer (ODAs) and inner dynein arms (IDAs), indicating that C11ORF70 is involved in cytoplasmic assembly of dynein arms. Expression analyses of C11ORF70 showed that C11ORF70 is expressed in ciliated respiratory cells and that the expression of C11ORF70 is upregulated during ciliogenesis, similar to other previously described cytoplasmic dynein arm assembly factors. Furthermore, C11ORF70 shows an interaction with cytoplasmic ODA/IDA assembly factor DNAAF2, supporting our hypothesis that C11ORF70 is a novel preassembly factor involved in the pathogenesis of PCD. The identification of a novel genetic defect that causes PCD and male infertility is of great clinical importance as well as for genetic counselling.

scholarly journals Higher throughput drug screening for rare respiratory diseases: Readthrough therapy in primary ciliary dyskinesia

2021 ◽  
pp. 2000455
Author(s):  
Dani Do Hyang Lee ◽  
Daniela Cardinale ◽  
Ersilia Nigro ◽  
Colin R. Butler ◽  
Andrew Rutman ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
High Throughput Screening ◽  
Primary Ciliary Dyskinesia ◽  
Respiratory Diseases ◽  
Beat Frequency ◽  
Screening Method ◽  
Respiratory Epithelium ◽  
Basal Cells ◽  
Translational Readthrough ◽  
Ciliary Dyskinesia

Development of therapeutic approaches for rare respiratory diseases is hampered by the lack of systems that allow medium-to-high-throughput screening of fully differentiated respiratory epithelium from affected patients. This is a particular problem for primary ciliary dyskinesia (PCD), a rare genetic disease caused by mutations in genes that adversely affect ciliary movement and consequently mucociliary transport. Primary cell culture of basal epithelial cells from nasal brush biopsies, followed by ciliated differentiation at air-liquid interface (ALI) has proven to be a useful tool in PCD diagnostics but the technique's broader utility, including in pre-clinical PCD research, has been restricted by the limited number of basal cells that it is possible to expand from such biopsies. Here, we describe an immunofluorescence screening method, enabled by extensive expansion of PCD patient basal cells and their culture into differentiated respiratory epithelium in miniaturised 96-well transwell format ALI cultures. Analyses of ciliary ultrastructure, beat pattern and beat frequency indicate that a range of different PCD defects can be retained in these cultures. As proof-of-principle, we performed a personalised investigation in a patient with a rare and severe form of PCD (reduced generation of motile cilia, RGMC), in this case caused by a homozygous nonsense mutation in the MCIDAS gene. The screening system allowed drugs that induce translational readthrough to be evaluated alone or in combination with nonsense-mediated decay inhibitors. Restoration of basal body formation in the patient's nasal epithelial cells was seen in vitro, suggesting a novel avenue for drug evaluation and development in PCD.

LSC Abstract – Oxidative stress in nasal epithelial cells from patients with primary ciliary dyskinesia

2016 ◽  
Author(s):  
Ana Reula ◽  
Shirley Camacho ◽  
Amparo Escribano ◽  
Sara Pastor ◽  
Silvia Castillo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Primary Ciliary Dyskinesia ◽  
Nasal Epithelial Cells ◽  
Ciliary Dyskinesia
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