Analiza variabilnosti gena za eritropoetin pri družinski eritrocitozi.

Prijatelj, Tinkara (author), Debeljak, Nataša (mentor), Kunej, Tanja (comentor)

Eritrocitoza zajema heterogeno skupino motenj, ki se odražajo v povečanem številu eritrocitov. Družinska eritrocitoza (DE) je prirojena motnja z različnim genetskim ozadjem. Mutacije v eritropoinskem receptorju so značilne za primarno DE tipa 1, medtem ko so za sekundarne DE tipa 2-4 odgovorne mutacije v genih vključenih v biološko pot zaznavanja kisika ter regulacijo izražanja eritropoetina (EPO). Cilj magistrske naloge je bil pregledati objavljeno literaturo na področju razvoja DE v povezavi z različicami zaporedja DNA v genu EPO, s pomočjo bioinformacijskih orodij napovedati nove biooznačevalce in eksperimentalno preveriti različice v genu EPO pri dveh bolnikih s sumom na DE. V objavljenih kliničnih študijah smo našli šest različic v genu EPO, povezanih z eritrocitozo ali povišanim hematokritom. Poenotili smo heterogeno nomenklaturo zapisa različic zaporedja DNA. Orodji SIFT in PolyPhen predvidita največji škodljiv vpliv na funkcijo proteina za dodatnih sedem različic gena EPO. Analiza nukleotidnega zaporedja dveh bolnikov s sumom na DE je potrdila prisotnost različice zaporedja DNA v ojačevalcu gena EPO, vendar bi bilo z dodatnimi raziskavami potrebno potrditi vzročnost različice za pojav DE. Metoda je bila prenesena v klinično uporabo. Poleg tega bi bilo potrebno opredeliti vpliv ostalih različic v regulatornih regijah gena. Standardizirano poročanje povezav genotip-fenotip bi olajšala primerjavo med študijami in hitrejši razvoj področja.

• Language: Slovenian
• Keywords: eritropoetin, gen, različice zaporedja DNA, določanje nukleotidnega zaporedja, eritrocitoza, družinska eritrocitoza
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [T. Prijatelj]
• Year: 2018
• PID: 20.500.12556/RUL-99897
• UDC: 601.4:577.12:606:616-056.7(043.2)
• COBISS.SI-ID: 8931961
• Publication date in RUL: 20.02.2018

Secondary language

• Language: English
• Title: Analysis of the erythropoietin gene variability in familial erythrocytosis

Abstract

Erythrocytosis is characterized by the expansion of erythrocyte compartment including elevated red blood cell number. Familial erythrocytosis (FE) is a congenital disorder with different genetic background. Type 1 FE is primary FE caused by mutation in erythropoietin receptor gene. Type 2-4 FE are secondary FEs caused by mutations of genes involved in oxygen sensing pathway important for erythropoietin (EPO) regulation. The aim of the master thesis was to: review published literature of associations between EPO gene variations and development of FE, prioritization of novel biomarkers using bioinformatics tools, and experimentally analyze EPO gene variations in two patients suspected of FE. We collected in published clinical studies six variants of the EPO gene associated with erythrocytosis or upper haematocrit. Heterogeneous nomenclature of sequence variants was unified. Seven additional EPO gene variants had the highest predicted deleterious effect on protein function using SIFT and PolyPhen. The sequence analysis of two related patients suspected of FE confirmed the presence of one variation in EPO enhancer, future studies should be performed to investigate causative for FE. The method was transferred into clinical use. Role of variants in regulatory regions should be further investigated. Standardized reporting format of genotype-phenotype associations would enable comparison between the studies and faster development of the study field.

• Keywords: erythropoietin, gene variations, sequencing, erythrocytosis, familial erythrocytosis