Vpliv polimorfizma rs642961 v regulatorni regiji gena IRF6 na nastanek orofacialnih shiz in prirojenih srčnih napak v slovenski populaciji : magistrski študijski program Laboratorijska biomedicina

Glavič, Marjetka (author), Karas Kuželički, Nataša (mentor), Šmid, Alenka (comentor)

Pogostost pojavljanja razvojnih nepravilnosti v splošni populaciji znaša 2–3 %, med najpogostejše pa sodijo srčne napake (CHD) in orofacialne shize (OFC). Po vzroku nastanka ločimo sindromske oblike, kjer se napaka pojavi kot eden od patogenetskih znakov bolezni, katere vzrok je bolj ali manj pojasnjen, in nesindromske, katerih etiologija nastanka je kompleksna. Pri slednjih gre večinoma za večfaktorska obolenja, kjer poleg genetskih faktorjev k povečanemu tveganju prispevajo tudi okoljski in demografski dejavniki. Eden od polimorfizmov, ki ga povezujejo s pojavom OFC, je rs642961, v regulatorni regiji gena IRF6. Namen magistrskega dela je bil ugotoviti, kako polimorfizem rs642961 vpliva na pojavnost OFC in CHD v slovenski populaciji, ob upoštevanju vpliva izbranih okoljskih in demografskih dejavnikov. V raziskavo smo vključili 179 otrok z OFC in njihovih mater, 150 otrok s CHD in njihovih mater ter 200 otrok brez klinično potrjene prirojene okvare in njihovih mater . V raziskavi sorojencev je dodatno sodelovalo 46 otrok brez klinično potrjene OFC, iz družin vključenih v prvem delu. S pomočjo reagenčnega kita smo izolirali DNA iz brisa bukalne sluznice in z uporabo TaqMan hidrolizirajočih sond določili genotip za IRF6 rs642961. Za pridobitev podatkov o demografskih in okoljskih dejavnikih so matere izpolnile anketni vprašalnik. S pomočjo statističnega programa smo analizirali spremenljivke, ki bi lahko imele vpliv na pojavnost CHD oz. OFC. Z modeli logistične regresije smo dokazali, da je za obe skupini malformacij pozitivna družinska anamneza največji dejavnik tveganja. Verjetnost za nastanek obeh je večja tudi v primeru prisotnosti drugih kroničnih bolezni pri materi in pri materah kadilkah. Povišana telesna temperatura v nosečnosti je bila značilen dejavnik tveganja le za pojav OFC. Dokazali smo, da vnos folatnih pripravkov v nosečnosti zmanjša pojavnost OFC in CHD, ne glede na obdobje jemanja. Statistično značilne razlike med testno (OFC oz. CHD) in kontrolno skupino v preiskovanem vzorcu za genotip polimorfizma rs642961 v genu IRF6 nismo dokazali. V študiji sorojencev smo ugotovili, da je bil delež otrok, ki imajo vsaj en mutiran alel za rs642961 v genu IRF6 za kar 40% višji v skupini otrok z OFC kot pri njihovih zdravih sorojencih (p = 0,004).

• Language: Slovenian
• Keywords: orofacialne shize prirojene srčne napake polimorfizem rs642961 študije sorojencev primeri s kontrolami
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FFA - Faculty of Pharmacy
• Place of publishing: Ljubljana
• Publisher: [M. Glavič]
• Year: 2018
• Number of pages: X, 88 str.
• PID: 20.500.12556/RUL-120169
• UDC: 577.12:616.12-039(043.3)
• COBISS.SI-ID: 4646001
• Publication date in RUL: 16.09.2020

Secondary language

• Language: English
• Title: The influence of polymorphism rs642961 in the regulator region of IRF6 gene on the occurrence of orofacial clefts and congential herat disease in the Slovenian population

Abstract

The incidence of congenital malformations in the general population is 2-3%, the most common are congenital heart disease (CHD) and orofacial clefts (OFC). In syndromic forms, where the error occurs as one of the pathogenetic signs of the disease, the cause of which is more or less known, whereas in non-syndromic, the etiology is complex. The latter are mainly multifactorial diseases, where, in addition to genetic factors, environmental and demographic factors also contribute to the increased disease risk. One of the polymorphisms associated with the occurrence of OFC is rs642961 in the regulatory region of the IRF6 gene. The purpose of the master's thesis was to determine how polymorphism rs642961 effects the occurrence of OFC and CHD in the Slovenian population, taking into account the influence of selected environmental and demographic factors. The study included 179 children with OFC and their mothers , 150 children with CHD and their mothers, and 200 children without clinically confirmed congenital malformation and their mothers . In addition, 46 children without clinically confirmed OFC from the families included in the first part were also recruited for the sibling study. Using the reagent kit, we isolated the DNA from the buccal mucosa and determined the genotype for IRF6 rs642961 using TaqMan hydrolysis probes. In order to obtain data on demographic and environmental factors, the mothers filled in a questionnaire. Using statistical analysis software, we analyzed all the variables that could have an impact on the incidence of CHD or OFC. With the logistic regression model, we have shown that the greatest risk for both OFC and CHD is a positive family history of the disease. The likelihood of both malfomations is even greater in mothers with other chronic diseases and in smoking mothers. Fever during pregnancy was a significant risk factor for the occurrence of OFC only. We have proven that the use of folate preparations in pregnancy protects from the occurrence of OFC and CHD, regardless of the duration of the treatment. Statistically significant differences between the test (OFC or CHD) and the control group in IRF6 rs642961 genotypes distribution were not detected. In a study of siblings, the proportion of children with at least one mutated allele for rs642961 in the IRF6 gene was significantly higher by as much as 40 % in the OFC children compared to their healthy siblings.

• Keywords: orofacial clefts congenital heart disease polymorphism rs642961 case-control study sibling study