Analiza vpliva genetskega polimorfizma v genu GRHL3 na razvoj orofacialnih shiz: študija primerov in kontrol ter študija družin

Bavčar, Manca (author), Karas Kuželički, Nataša (mentor), Urbančič, Dunja (comentor)

Orofacialne shize (ang. orofacial clefts; OFC) so prirojene nepravilnosti obraza, ki so posledica napačnega razvoja ustnice, mehkega in/ali trdega neba, nosne pregrade ter čeljustnega grebena. Posamezniki se zaradi prirojene nepravilnosti soočajo z estetskimi, zdravstvenimi, funkcionalnimi in psihološkimi izzivi, ki zahtevajo doživljenjsko multidisciplinarno zdravstveno oskrbo. Pojavnost orofacialnih shiz se giblje od 1 na 2500 do 1 na 500 živorojenih otrok in se razlikuje glede na spol, raso ter etnično pripadnost. Na nastanek orofacialnih shiz vpliva preplet genetskih, demografskih in okoljskih dejavnikov, kar so v preteklosti dokazali v več študijah. V magistrski nalogi smo se osredotočili na proučevanje spremembe enega nukleotida v genu GRHL3, polimorfizma rs41268753 C>T, v povezavi z razvojem orofacialnih shiz pri posamezniku. Proučevali smo 303 posameznike znotraj 125 različnih družin, kjer je eden od otrok imel prisoten razcep v orofacialnem področju. Izvedli smo študijo primerov in kontrol, in v kontrolno skupino vključili 137 zdravih odraslih preiskovancev. Iz vseh vzorcev smo izolirali DNA ter za ugotavljanje genotipa posameznikov uporabili genotipizacijo s hidrolizirajočimi sondami, ki temelji na verižni reakciji s polimerazo. Ugotovili smo, da je frekvenca variantnih alelov v slovenski populaciji pričakovano enaka kot v splošni evropski populaciji. Pokazali smo, da je frekvenca variantnih alelov T znotraj skupine bolnikov z orofacialno shizo signifikantno višja od frekvence variantnih alelov T v kontrolni skupini zdravih preiskovancev, kar bi lahko nakazovalo na vpliv genetskega polimorfizma rs41268753 na razvoj OFC. V šestih različnih družinah, kjer se je orofacialna shiza pojavila v več generacijah, smo izvedli študijo družin, izrisali družinska drevesa in proučili, ali variantni alel T segregira s patološkim fenotipom v družinah, kjer se pojavlja OFC. Naše ugotovitve so pokazale, da polimorfizem rs41268753 ne vpliva na razvoj shiz po monogenskem načinu dedovanja, ampak gre verjetno za multifaktorski vpliv, kjer igrajo pomembno vlogo tudi drugi genetski, okoljski in demografski dejavniki.

• Language: Slovenian
• Keywords: orofacialne shize, genetski polimorfizem, gen GRHL3, študija primerov in kontrol, študija družin.
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2025
• PID: 20.500.12556/RUL-167964
• Publication date in RUL: 21.03.2025

Secondary language

• Language: English
• Title: Analysis of the effect of the genetic polymorphism in the GRHL3 gene on the development of orofacial clefts: a case-control study and a family study

Abstract

Orofacial clefts (OFC) are congenital abnormalities of the face that result from incorrect development of the lips, soft and/or hard palate, nasal septum, and jaw ridge. Due to the congenital abnormality, individuals face aesthetic, health, functional, and psychological challenges that require lifelong multidisciplinary medical care. The incidence of orofacial clefts ranges from 1 in 2,500 to 1 in 500 live births and varies by gender, race, and ethnicity. The formation of orofacial clefts results from the interplay between genetic, demographic, and environmental factors, which has been proven in the past in several studies. In the master's thesis, we focused on the single nucleotide changes in the GRHL3 gene, the polymorphism labeled as rs41268753 C>T, in connection with the development of orofacial clefts in affected individuals. We studied 303 individuals within 125 different families, where one of the children had a cleft in the orofacial area. We conducted a case-control study and included 137 healthy adult subjects as a control group. We isolated DNA from all samples and determined the genotype of the study subject by hydrolysis probes and polymerase chain reaction. We found that the frequency of variant alleles in the Slovenian population is expected to be the same as in the general European population. We have shown that the frequency of variant T alleles within the group of patients with orofacial clefts is significantly higher than the frequency of variant T alleles in the control group of healthy subjects, which could indicate the influence of the genetic polymorphism rs41268753 on OFC development. Additionally, we performed a family study in six different families where orofacial clefts occurred in several generations. We created family trees and examined whether the variant allele T segregates with the pathological phenotype in families with OFC. Our findings show that the rs41268753 polymorphism does not trigger the development of orofacial clefts through a monogenic inheritance model. The onset of the defects is probably multifactorial, with other genetic, environmental, and demographic factors playing an important role in its formation.

• Keywords: orofacial clefts, genetic polymorphism, gene GRHL3, case-control study, family study.