GAUCHER HASTALIĞININ TANISI VE ŞİDDETİNİN GÖSTERGESİ OLARAK PTERİDİN METABOLİTLERİNİN ARAŞTIRILMASI
Project no. 20414 · Status: Kapanmış (Yayın bekleniyor) · 2019
Summary
The aim of this project is to investigate the potential benefit of levels of pteridine metabolites in the diagnosis and monitoring of Gaucher's disease. Pteridines and their derivatives function as intermediates in the metabolism of various vitamins and cofactors, and their relevance to disease has inspired new research to study their role as disease biomarkers. For this purpose 14 patients with a diagnosis of lysosomal Gaucher who were being followed up in the policlinic of Ege University Faculty of Medicine Child Health and Diseases Department of Metabolism were included in the study. While blood samples were taken from patients for routine monitoring samples were also collected and studied for this study. Pterine metabolites and neopterin levels in blood and urine were determined by the LC MS/ MS-based method. Representative chromatograms obtained as a result of optimization of LC and MS/MS conditions, the peaks were symmetrical, noise and tailing were almost nonexistent, and signal/noise ratio was high. The R2 values of the linear range graphs obtained for each metabolite were found to be 0.995. In the repeatability findings; CV% values are below 15% for high level both within day and between days. Urine Neopterin levels were determined with both ELISA kit and LC MS/MS methods. In the comparison of the two methods, both methods showed positive correlation. When the chitotriosidase enzyme activities of Gaucher patients were compared with the control group, CHIT levels of the patients were found significantly higher (p <0.001) than those of control grourp. While Neopterin levels were found to be higher in the urine of the patients compared to the healthy individuals, the tetrahydrobiopterin levels were found to be low. Dihydropteridine and 7-biopterin (primapterin) levels were also found higher in patients but this elevation was not statistically significant. Considering the neopterin/tetrahydrobiopterin ratio, this ratio was found to be significantly higher in patients compared to healthy individuals. When the ROC curve was drawn, it was determined that Neopterin alone did not have sufficient sensitivity as a disease indicator while the Neopterin/BH4 ratio showed similar specificity and sensitivity with chitotrioidase. Chitotriosidase activity in healthy individuals did not show any correlation with pterin metabolites while chitotriosidase activity in patients was positively correlated with urinary neopterin (p = 0.018, r = 0.622) and 7-biopterin (p = 0.018, r = 0.622) levels. High levels of neopterin in the urine of Gaucher patients indicate that NP may be a reliable marker that can be used in screening and prognosis of the disease in these patients. High neopterin levels and low BH4 levels were evaluated as macrophage activation indicators. The most important finding obtained in this study is that the Neopterin/BH4 ratio was found to be significantly high in the urine of Gaucher patients and showed a sensitivity equivalent to Chitotriosidase. Our data suggests that the determination of BH4 an Neopterin levels in urine by LC MS/MS will be particularly useful in patients with genetically low chitotriosidase. We think that the Neopterin/BH4 ratio can be used in the clinic as an important indicator because the urine sample can be obtained non-invasively, the results can be obtained quickly with LC MS / MS and the accuracy and precision of the demonstrated test.