TY - JOUR T1 - Evaluation of gene expression during in vitro ciliogenesis to search for novel candidate genes in primary ciliary dyskinesia JF - European Respiratory Journal JO - Eur Respir J VL - 42 IS - Suppl 57 SP - P529 AU - Tamara Paff AU - Eric Haarman AU - Hans Daniels AU - Youssef Moutaouakil AU - Dimitra Micha AU - Maarten Massink AU - Piet Postmus AU - Gerard Pals Y1 - 2013/09/01 UR - http://erj.ersjournals.com/content/42/Suppl_57/P529.abstract N2 - IntroductionMutations in genes that are important in ciliary motility often cause primary ciliary dyskinesia (PCD). In half of the patients, causal mutations in any of the eighteen known PCD genes cannot be identified.AimThe aim of this pilot study is to investigate gene expression in respiratory epithelial cells during ciliogenesis. We hypothesized that a) all known PCD genes show similar expression profiles and that b) clustering genes with similar expression may provide novel PCD candidate genes.MethodsWe collected nasal epithelial curette biopsies from 2 healthy controls. Cells were cultured using a monolayer-suspension cell culture (Willems,T et al. J Cyst Fibros 2004). RNA was isolated at three time points: at the end of the monolayer culture (no cilia), after three days in suspension culture (growing cilia) and at the end of the suspension culture (full grown cilia). Whole genome expression arrays were performed and differential gene expression was analyzed by ANOVA.ResultsExcept for TXNDC3, we observed a time dependent increase in expression levels in all known PCD genes. However, for HEATR2 and CCDC40 this difference did not reach statistical significance. A total of 1511 genes were significantly up-regulated during ciliogenesis. This cluster was highly enriched for genes related to cilia.ConclusionWe identified a cluster of 1511 genes that showed an increase in expression levels during in vitro ciliogenesis, including 15 of the 18 known PCD genes. Using this novel application on a larger sample size may provide a more accurate cluster of possible PCD candidate genes. ER -