magistrsko delo
Tina Turel (Avtor), Nataša Debeljak (Mentor), Marina Klemenčič (Član komisije za zagovor), Marko Novinec (Član komisije za zagovor)

Povzetek

Eritrocitoza je redka pridobljena ali prirojena bolezen, za katero je značilno povišano število rdečih krvničk. Prirojene eritrocitoze (ECYT1-8) so skupina bolezni z genetsko spremembo v enem izmed genov, ki so vključeni v proces regulacije eritropoeze. S prirojeno eritrocitozo je do zdaj neposredno povezanih devet genov, in sicer EPOR, VHL, EGLN1, EPO, EPAS1, HBB, HBA1, HBA2 in BPGM. V razvoj pridobljene eritrocitoze sta vključena gena JAK2 in SH2B3, neopredeljena pa ostaja njuna vloga pri razvoju prirojenih eritrocitoz. Kljub sekvenčni analizi kodirajočih regij genov, vključenih v eritrocitozo, ostaja 70 % bolnikov s sumom na prirojeno eritrocitozo neopredeljenih. Namen magistrske naloge je bil določiti genetske različice v nekodirajočih regijah 11 genov, ki vplivajo na razvoj eritrocitoze ter potrditi njihovo prisotnost v genomih bolnikov. V prvem sklopu naloge smo z bioinformatskimi orodji analizirali nekodirajoče regije 11 genov. Pri pregledu literature smo določili genomske lokacije nekodirajočih regij, lokacije histonov, mesta metilacij in genomske lokacije CpG otočkov. Pridobljene podatke smo primerjali z rezultati analize NGS iz predhodne raziskave na bolnikih ter določili genetske različice, ki bi lahko vplivale na razvoj prirojene eritrocitoze. Pri nadaljnji analizi smo genetskim različicam določili oznako (ID SNP), povezavo z eritrocitozo oziroma povečanim hematokritom, sovpadanje z regijo kriptičnega eksona ter predikcijo patogenosti CADD. Izmed izbranih genetskih različic smo ID SNP lahko določili samo sedmim genetskim različicam. Dve od teh sta bili povezani s povišanim hematokritom in povišanim izražanjem EPO. Ker je CADD pri vseh genetskih različicah nižji od 20, kar je meja za patogenost, večine genetskih različic nismo vključili v nadaljnjo eksperimentalno analizo. V drugem sklopu naloge smo v eksperimentalno analizo vključili genetski različici rs551238 in rs1617640 gena EPO, ki sta bili predhodno povezani s povišanim hematokritom. Izvedli smo genotipizacijo na 26 DNA vzorcih bolnikov. Rezultati so pokazali na morebitno povezavo med analiziranima SNP, saj so se genotipi pri posameznem vzorcu ujemali. Na podlagi določitve genotipa enega SNP smo lahko napovedali tudi genotip drugega. Analiza genotipov družinskih članov je nakazala avtosomno dominantno dedovanje.

Ključne besede

eritrociti;prirojena eritrocitoza;eritropoeza;hormon eritropoetin;EPO;nekodirajoče regije;genotipizacija;magistrska dela;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.09 - Magistrsko delo
Organizacija: UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik: [T. Turel]
UDK: 577.21:616.15(043.2)
COBISS: 90752515 Povezava se bo odprla v novem oknu
Št. ogledov: 204
Št. prenosov: 34
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Influence of non-coding regulatory regions of erythropoiesis-related genes on the development of congenital erythrocytosis
Sekundarni povzetek: Erythrocytosis is a rare, acquired or natural disease characterized by an increased number of red blood cells. Congenital erythrocytosis (ECYT1-8) is a group of diseases with a genetic change in one of the genes involved in the process of regulating erythropoiesis. So far, nine genes have been directly linked to congenital erythrocytosis, namely EPOR, VHL, EGLN1, EPO, EPAS1, HBB, HBA1, HBA2 and BPGM. The JAK2 gene in SH2B3 is involved in the development of acquired erythrocytosis, but their role in the development of congenital erythrocytosis remains undefined. In addition, sequential analysis of the coding regions of the genes involved in erythrocytosis leaves 70% of patients with suspected natural erythrocytosis undecided. The purpose of the master's thesis was to determine the genetic differences in the non-coding regions of 11 genes that influence the development of erythrocytosis and to confirm their presence in the genomes of patients. In the first part of the task, we analyzed the non-coding regions of 11 genes with bioinformatics tools. In reviewing the literature, we determined the genomic locations of noncoding regions, histone locations, methylation sites, and genomic locations of CpG islands. The obtained data were compared with the results of NGS analyzes from previous studies on patients and genetic variants that could influence the development of the nature of erythrocytosis were determined. In addition, we analyzed genetic variants determined by tag (ID SNP), association with erythrocytosis, or increased hematocrit, matching with the cryptic exon region and CADD pathogenicity prediction. Of the selected genetic variants, only seven genetic variants could be assigned to the SNP ID. Two of these were associated with elevated hematocrit and elevated EPO expression. Because CADD is lower than 20 in all genetic variants, which is the limit for pathogenicity, most genetic variants were not included in further experimental analysis. In the second part of the task, genetic variants of rs551238 and rs1617640 of the EPO gene, which were previously associated with elevated hematocrit, were included in the experimental analysis. Genotyping was performed on 26 sample DNA samples. The results indicated a possible association between the analyzed SNPs, as the genotypes matched in each sample. Based on the determination of the genotype of one SNP, we were also able to predict the genotype of the other. Analysis of genotypes of family members indicated autosomal dominant inheritance.
Sekundarne ključne besede: congenital erythrocytosis;non-coding regions;EPO;genotyping;
Vrsta dela (COBISS): Magistrsko delo/naloga
Študijski program: 1000377
Konec prepovedi (OpenAIRE): 1970-01-01
Komentar na gradivo: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, smer Biokemija
Strani: II, 70 str.
ID: 13826372