20.500.12556/DKUM-44552 ASOCIACIJSKA ANALIZA ZA POTRDITEV KANDIDATNIH DNA POLIMORFIZMOV NA KROMOSOMU 2 IN 22 PREDHODNO POVEZANIH S SLOVENSKIMI BOLNIKI S CROHNOVO BOLEZNIJO Z UPORABO MIKROMREŽE ICHIP ASSOCIATIVE ANALYSIS TO CONFIRM THE CANDIDATE DNA POLYMORPHISMS ON CHROMOSOME 2 AND 22 PREVIOUSLY ASSOCIATED WITH THE SLOVENIAN PATIENTS WITH CROHN'S DISEASE USING MICROARRAYS ICHIP Povzetek Kronična vnetna črevesna bolezen (KVČB) je kompleksna bolezen. Znana sta dva glavna podtipa in sicer, Crohnova bolezen (CB) ter ulcerozni kolitis (UK). Zakaj bolezen nastane, ni popolnoma jasno, so pa študije pokazale, da k razvoju bolezni prispevajo okoljski in genetski dejavniki. Veliko število polimorfizmov posameznega nukleotida (SNP) je vpleteno v mehanizem bolezni. Predhodno je bila s pomočjo mikromreže (»Immunochip«) izvedena genska tipizacija za približno 200 000 polimorfizmov pri 250 slovenskih bolnikih s KVČB ter prav tako 250 kontrol zdravih posameznikih. Rezultat asociacijske analize je na kromosomu 2 prikazal 11 neodvisno razporejenih lokusov na katerih je 332 SNP-jev s p vrednostjo p<0,05 ter na kromosomu 22, kjer je prav tako 5 neodvisno razporejenih lokusov ter 101 SNP-ja jev s p vrednostjo p<0,05. Ker lahko pri multiplem testiranju pride do pozitivno lažnih rezultatov, smo v raziskavi za potrditev kandidatnih polimorfizmov izvedli potrditveno asociacijsko analizo na dodatnih bolnikih ter zdravih kontrolah. Namen diplomskega dela je bil ugotoviti ali so izbrani polimorfizmi na kromosomu 2 in 22 povezani s slovensko populacijo bolnikov s KVČB. Tako smo v raziskavi izbrali 4 SNP-je, tri na kromosomu 2 in 1 na kromosomu 22 ter izvedli genotipizacijo z metodo HRM na 482 vzorcih bolnih in 350 vzorcih zdravih kontrolah posameznikov. V nalogi smo potrdili povezavo med vsemi analiziranimi SNP-ji. V primeru alela C (p=0,001; OR= 1,578; CI(95%) = 1,200-2,076) za SNP rs6733174 smo odkrili, da veča tveganje za razvoj KVČB (p=0,001; OR= 1,578; CI(95%) = 1,200-2,076). SNP rs137618 ima povečano tveganje za razvoj bolezni v primeru alela C (p= 0,007; OR= 0,678; CI(95%)= 0,512-0,898) ter smo tudi odkrili, da homozigoti tega istega SNP-ja z genotipom C/C (p= 0,00159; OR= 2,070; CI(95%) = 1,313-3,262), povečujejo tveganje za KVČB (p= 0,00159; OR= 2,070; CI(95%) = 1,313-3,262). Do enakega zaključka smo prišli pri pregledu rezultatov za SNP rs7600901. V primeru alela G (p= 0,000347; OR= 1,778; CI(95%) = 1,778-2,441), le-ta nosi povišano tveganje za razvoj bolezni (p= 0,000347; OR= 1,778; CI(95%) = 1,778-2,441). Zanimivo pa je to, da smo tudi za SNP rs7590126 (gen PAPOLG), našli signifikantne povezave s KVČB v primeru alela C (p=0,026; OR= 1,476; CI(95%) = 1,051-2,073) . Tako smo potrdili statistično najbolj signifikantno povezavo ravno pri bolnikih s CB. KVČB še tako vedno ostaja dokaj »neprepoznavna« bolezen, saj splošni zdravniki zelo pozno ali pa sploh ne prepoznajo osnovnih simptomov. Zato je smiselno področje genetike KVČB raziskovati še naprej, kajti z novo odkritimi polimorfizmi SNP, povezanimi z boleznijo, pripomoremo k hitrejšemu odkrivanju bolezni, razvoju novih zdravil, natančnejšemu napovedovanju poteka bolezni ter uspešnejšemu zdravljenju, hkrati pa lahko preprečimo marsikatere težave, ki bi nastale pri zdravljenju bolezni. Abstract Inflammatory bowel disease (IBD) is a complex disease. Known are two major subtypes namely, Crohn's disease (CB) and ulcerative colitis (UK). Why the disease occurs is not entirely clear, but studies have shown that the disease contribute to the development of environmental and genetic factors. A large number of single nucleotide polymorphisms (SNP), is involved in the mechanism of the disease. Previously, with the help of microarrays ("Immunochip") genotyping of approximately 200 000 polymorphisms in 250 Slovenian patients were conducted, as well as 250 controls of healthy individuals. The results from the associative analysis on chromosome 2 showed 11 locus on wich 323 SNP's with a p value of p<0,05 and on chromosome 22, from wich are 5 locus and 101 SNP's with a p value of p<0,05. Since the multiplex test may show false positive results, we conducted a associative polymorphisms check on the patient in order to confirm the study . As well as on sick and helathy individuals. The purpose of diploma work was to determine if the polymorphisms on the chromosome 2 and 22 are connected with the patients among the slovene population, that have chronic inflammatory bowel disease. Thus, we chose to study four SNPs, three on chromosome 2 and one on chromosome 22 and performed genotyping by HRM method on 482 samples of diseased and 350 healthy controls patterns of individuals. In the study, we confirmed the link between all the analyzed SNPs. In the case of allele C (p = 0,001; OR = 1,578; CI (95 % ) = 1,200 – 2,076) of the SNP rs6733174 was found to increase the risk for development of IBD (p = 0,001; OR = 1,578; CI ( 95 % ) = 1,200 – 2,076). SNP rs137618 has an increased risk of developing the disease in the case of allele C (p = 0,007; OR = 0.678 ; CI ( 95 % ) = 0,512-0,898) , and we have also found that this same homozygote SNP genotype C/C (p = 0,00159; OR = 2,070; CI (95%) = 1,313 – 3,262) , increasing the risk of IBD ( p = 0,00159; OR = 2,070; CI (95%) = 1,313 – 3,262). To the same conclusion we came in the review of the results for SNP rs7600901. In the case of the G allele (p = 0,000347; OR = 1,778; CI ( 95 % ) = 1,778 – 2,441), the latter to bear an increased risk for the development of the disease ( p = 0,000347; OR = 1,778 ; CI ( 95 % ) = 1,778 – 2,441) . It is interesting that we have for SNP rs7590126 (gene PAPOLG) also found significant relationship with IBD in the case of C allele (p = 0,026; OR = 1,476; CI (95%) = 1,051 – 2,073). Thus, we confirmed the statistically most significant link straight in patients with CB. IBD has thus remains quite "unrecognizable" disease because general practitioners very late or not at all identify basic symptoms. So it makes sense to explore the genetics of IBD continue, because the newly discovered SNP polymorphisms associated with the disease, contributing to more rapid detection of disease, the development of new drugs, more accurate prediction of the disease and successful treatment, that we can prevent many problems that would arise in the treatment of diseases. Analiza kromosomov pri slovenskih bolnikih s kronično vnetno črevesno boleznijo. Analysis of chromosomes in Slovenian patients with inflammatory bowel disease. true true false [E. Friedl] Slovenski jezik Angleški jezik Diplomsko delo 2014-06-05 09:25:50 2014-06-27 10:35:01 2022-05-13 03:06:38 2021-05-01 03:27:44 2014 0 Maribor 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616.35-002:616.253(497.4):168.2:576.316 24655160 URN:SI:UM:DK:AQVFIRMD VS_Friedl_Evelina_2014.pdf VS_Friedl_Evelina_2014.pdf 1 0019A78109992196CF2AC4776A283650 42cc227a276ac523862adaa5a8929273e9d8a82167a109706018cfac2bacdef6 e1cbb577-7c0a-11eb-bb7a-00155d0001ca https://dk.um.si/Dokument.php?lang=slv&id=64008 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0