Gene expression during ciliagenesis: Towards a novel candidate gene list for Primary Ciliary Dyskinesia.

Primary Ciliary Dyskinesia (PCD) is a hereditary disorder occurring in 1:15.000-30.000 live births, with increased frequency in genetically isolated populations. PCD is characterised by dyskinesia of epithelial cilia causing chronic respiratory disease. Diagnosing patients can be challenging, as there is no gold standard test. Due to the many unknown disease causing gene defects, genetic testing is still not possible in most cases. To this date, our understanding of the molecular composition of cilia is far from complete. Investigating which genes are important in cilia genesis will contribute to a more complete candidate gene list and development of a diagnostic test for Primary Ciliary Dyskinesia.



Objective of the study:

We aim to assess gene expression patterns in cilia-producing cells from healthy controls to develop a candidate gene list for PCD.



Study design:

This observational study will be conducted at the VU University Medical Center. Nasal curette and brush biopsies will be performed on healthy volunteers to obtain respiratory epithelial cells. Cells will be cultured and RNA will be isolated at three different time points during cilia genesis. Subsequently, Affymetrix gene expression arrays will be performed on cDNA and analyzed by cluster analysis.



Study population:

Six healthy volunteers (employees from the Dept. of Pulmonary Medicine/Pediatrics/Clinical Genetics) will be recruited to participate in this study.



Primary study parameters/outcome of the study:

Gene expression levels and pattern.



Secundary study parameters/outcome of the study:

Cell culture success rate with brush biopsies.

We aim to assess gene expression patterns in cilia-producing cells from healthy controls to develop a candidate gene list for PCD. We hypothesise that:

1. Known PCD genes show similar gene expression patterns during ciliagenesis;

2. Novel candidate genes can be identified by clustering genes with similar expression patterns.

RNA will be isolated at three different time points during ciliagenesis (before, during and after the growth of cilia).

Nasal curette and brush biopsies will be performed on healthy volunteers to obtain respiratory epithelial cells. Cells will be cultured and RNA will be isolated at three different time points during cilia genesis. Subsequently, Affymetrix gene expression arrays will be performed on cDNA and analyzed by cluster analysis.