Distinct pattern of lung gene expression in theCftr-KO mice developing spontaneous lung disease compared with their littermate controls

Abstract

Cystic fibrosis (CF) is caused by a defect in the CF transmembrane conductance regulator (CFTR) protein that functions as a chloride channel. Dysfunction of the CFTR protein results in salty sweat, pancreatic insufficiency, intestinal obstruction, male infertility, and severe pulmonary disease. Most of the morbidity and mortality of CF patients results from pulmonary complications. Differences in susceptibility to bacterial infection and variable degree of CF lung disease among CF patients remain unexplained. Many phenotypic expressions of the disease do not directly correlate with the type of mutation in the Cftr gene. Using a unique CF mouse model that mimics aspects of human CF lung disease, we analyzed the differential gene expression pattern between the normal lungs of wild-type mice (WT) and the affected lungs of CFTR knockout mice (KO). Using microarray analysis followed by quantitation of candidate gene mRNA and protein expression, we identified many interesting genes involved in the development of CF lung disease in mice. These findings point to distinct mechanisms of gene expression regulation between mice with CF and control mice.

Oxidative stress modulates the expression of genes involved in cell survival in ΔF508 cystic fibrosis airway epithelial cells

“…3) by RT-qPCR. Furthermore, many of the upregulated genes identified in our study, such as IL-6, IL-8, IL-1B, and IL-1A, have also been identified in other CF tissues (19,23,36,53). However, in recent experiments on CFBEs, the authors report no compelling evidence that mutations in CFTR induce a hyperinflammatory response in CF epithelial cells (20).…”

“…Several investigations of CF mouse models have reported overexpression of inflammatory genes in CF lungs (19,23,53) and intestinal tissue (36). However, these observations could not be consistently reproduced in CF bronchial epithelial cell lines or primary culture of CF epithelial cells (20).…”

Fenretinide prevents the development of osteoporosis in Cftr-KO mice

“…Overall, CF mice still represent the most useful and costefficient animal model available to study such a multifaceted and devastating disease. We have previously reported that the Cftr-KO mouse model develops spontaneous inflammatory lung disease [39][40][41][42]. In addition, we have recently demonstrated that they also exhibit an imbalance between DHA and AA, similar to CF patients [29].…”

The tetraspanin Tm4sf3 is localized to the ventral pancreas and regulates fusion of the dorsal and ventral pancreatic buds

“…As we had only two replicates for each sample (too small a number to use methods based on analysis of variance or integral methods, such as RMA or PLIER), we employed the method of consecutive sampling and coincidence test (Guilbault et al, 2006;Novak et al, 2006a;Novak et al, 2006b;Novak et al, 2002). In two-array comparisons, the genes are ordered according to mean signal intensity and grouped in bins containing n consecutive genes (in the present case, n=25).…”

“…Four different samples, two dorsal and two ventral, were reverse transcribed, labeled and hybridized to the Xenopus Affymetrix Genechip. Results were analyzed using the Affymetrix Expression Console and MAS5 algorithm, and because we only had two replicates, we employed the method of consecutive sampling and coincidence test (Guilbault et al, 2006;Novak et al, 2006a;Novak et al, 2006b;Novak et al, 2002). The results of this analysis yielded 158 genes as being ventral enriched and 68 genes as being dorsal enriched (see Tables S1 and S2 in the supplementary material).…”