CFTR Gene

CF is one of the most common autosomal recessive diseases in Caucasians (~1 per 2,500 births, 1/1,500 in Ireland). It has long known to be associated with defects in chloride electrolyte metabolism. Chloride sweat test can be used to identify affected patients by administering pilocarpine and a mild electric current to make a part of the skin sweat. In children, a sweat chloride concentration of more than 60 mmol/L suggests the possibility of CF; the level in adults is 80 mmol/L. The median age at death in Ireland: 25 years (in 2004, 17 in 2000) compared to 30 in U.K. The genetic defect underlying cystic fibrosis disrupts the functioning of several organs by causing ducts or other tubes to become clogged usually by thick, sticky mucus or other secretions. Clogging and infection of bronchial passages impede breathing. The infections progressively destroy the lungs. The only definitive treatment is lung transplantation, although physiotherapy can help offset severity of symptoms. Plugging of small bile ducts impedes digestion and disrupts liver function in perhaps 5% of patients. Occlusion of ducts prevents the pancreas from delivering critical digestive enzymes to the bowel in 85% of patients. Diabetes can result as well. Obstruction of the gut by thick stool necessitates surgery in about 10% of newborns. Absence of fine ducts, such as the vas deferens, renders 95% of male infertile. Occasionally, females are made infertile by a dense plug of mucus that blocks sperm from entering the uterus. The search for the CF gene was highly competitive throughout the 1980s. Tsui and his group managed to map the CF gene to chromosome 7 in 1985 which was further narrowed to 7q31 by identifying 2 flanking markers. 18 Restriction fragment length polymorphisms (RFLPs) were identified in a region of chromosome 7. The recombination fraction between these markers was low therefore the majority of markers were uninformative. Some markers were in linkage disequilibrium. Potential genes were searched for by comparing the sequence with other organisms and 3 conserved segments were obtained. One was expressed in the cDNA library from the sweat glands of affected patients. The sequence identified was small (113bp) and required further analysis to clone the entire gene. The final product protein, CFTR, predicted to contain 1,480 amino acids. Approximately 70% of CG patients had a deletion of a single codon (phenyalanine at position 508). The remaining 30% of CF mutations have slowly been unravelled. Greater than 1300 variations are now known. In Ireland, 8 mutations (including F508 deletion) account for 83% of the defective genes variants in the population. This high number of possible mutations makes it difficult to detect carriers with 100% accuracy. The standard test involves performing a heel prick with screening for levels of immunoreactive trypsin levels in serum. This is followed by a sweat and DNA test. Most DNA tests screen for the 25 most common mutations (8090% of Caucasians in the US). Current treatments involve nutritional repletion by a change in diet and pancreatic enzyme supplements, relief of airway obstruction by postural drainage, mechanical vests and recombinant Dnase, treatment of airway infection through oral and intravenous antibiotics, suppression of inflammation using steroids or high-dose ibuprofen or lung transplantation. Future treatments involve ion channel drugs that aim to block the Na+ channels, stimulate alternative CaCC channels or

increase Cl- secretion by stimulating K+ channels. Other treatments target protein repair such as suppression of premature stop codons through the use of aminoglycoside antibiotics, chemical, molecular or pharmacological chaperones to stabilise protein structure and protein folding, increase overall cell surface content, or stimulate existing channels with potentiators.