Uloga polimorfizama gena CES1 u kliničkoj medicini

master's thesis

• Title (english): The role of CES1 gene polymorphisms in clinical medicine
• Author: Ivana Prpić
• Mentor: Dunja Rogić (mentor)
• Mentor: LIVIJA ŠIMIČEVIĆ (mentor)
• Committee member: Dunja Rogić (predsjednik povjerenstva)
• Committee member: LIVIJA ŠIMIČEVIĆ (član povjerenstva)
• Committee member: Lidija Bach-Rojecky (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; Zagreb
• Defense date and country: 2023-09-22, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Dabigatran je aktivni oblik prolijeka dabigatraneteksilata koji pripada skupini direktnih oralnih antikoagulansa (DOAK)
te djeluje kao izravni oralni inhibitor trombina. Ranija istraživanja su pokazala značajnu interindividualnu varijabilnost
u plazmatskim koncentracijama dabigatrana zbog čega je u kasnijim studijama istraživan utjecaj genetičkih i
negenetičkih čimbenika na farmakokinetiku dabigatrana i kliničke ishode liječenja. Provođenjem cjelogenomske studije
povezanosti otkriveno je nekoliko polimorfizama jednog nukeotida (SNP) dva gena, CES1 i ABCB1, za koje do danas
postoji najviše dokaza kako njihove genetičke varijante utječu na farmakokinetiku dabigatrana i potencijalno mogu
dovesti do pojave neželjenih štetnih učinaka. Stoga je cilj ovoga istraživanja istražiti utjecaj dva polimorfizma CES1
(rs2244613 i rs8192935) na kliničke ishode terapije dabigatranom, točnije vidjeti doprinose li povećanom riziku od
neželjenih učinaka. Osim genetičkih čimbenika, istraživan je i utjecaj negenetičkih čimbenika te interakcija lijek-lijek.
Istraživanje je provedeno kao dio uspostavnoga istraživačkog projekta „Uloga farmakogenomike u predviđanju
nuspojava kardiovaskularnih lijekova (PGx CardioDrug)“ Hrvatske zaklade za znanost (HRZZ-UIP-2020-02-8189).
Genotipizacija istraživanih polimorfizama provedena je TaqMan® metodom lančane reakcije polimeraze u stvarnom
vremenu na ABI 7500 Real-Time PCR System uređaju. Na temelju medicinske dokumentacije ispitanika prikupljeni su
klinički značajni parametri poput dobi, spola, doze lijeka, indeksa tjelesne mase, komorbiditeta, eGFR, serumske
koncentracije kreatinina i konkomitantne terapije. U istraživanju je sudjelovalo 96 ispitanika (41 žena i 55 muškaraca)
u dobi od 21 do 93 godine na terapiji dabigatraneteksilatom. Šest ispitanika je isključeno iz statističke analize na temelju
isključnih kriterija. Krvarenje se pojavilo u 27 ispitanika (30 %), a tromboembolijski događaj u osam ispitanika (8,9 %).
Polimorfizam CES1 rs2244613 isključen je iz statističke obrade zbog odstupanja od Hardy-Weinbergove ravnoteže.
Statističkom analizom nije pronađena značajna povezanost polimorfizama CES1 rs8192935 i rizika od krvarenja,
odnosno tromboembolijskih događaja što je u skladu s rezultatima drugih ranije provedenih istraživanja. S druge strane,
pronađena je statistički značajna povezanost ženskoga spola, niže doze lijeka (110 mg) i oštećene bubrežne funkcije te
interakcija dabigatrana i lijekova koji potencijalno smanjuju njegovu učinkovitost s rizikom od krvarenja. S obzirom
kako je glavno ograničenje ovoga istraživanja premali broj ispitanika, u budućnosti je potrebno provesti opsežniju
studiju s većim uzorkom kako bi se potvrdila povezanost genetičkih varijanti CES1 s kliničkim ishodima terapije i
potencijalna klinička korist genetičkih testiranja s ciljem prepoznavanja rizičnih skupina pacijenata i optimizacije
terapije.

Abstract (english)

Dabigatran is an active form of prodrug dabigatran etexilate and acts as an oral direct thrombin inhibitor which is why
it belongs to the novel group of anticoagulants called direct oral anticoagulants (DOACs). Previous research showed
notable interindividual variability in plasma levels of dabigatran which is why the effect of genetic and nongenetic
factors on the pharmacokinetics of dabigatran and clinical outcomes was investigated. Genome-wide association studies
revealed several single nucleotide polymorphisms (SNPs) of two genes, CES1 and ABCB1, for which there is the most
evidence that their genetic variants affect the pharmacokinetics of dabigatran and can potentially lead to adverse effects.
The aim of this research is to investigate the impact of CES1 polymorphisms (rs2244613 and rs8192935) on the clinical
outcomes of dabigatran therapy. Besides genetic factors, we also studied the impact of nongenetic factors and drug-drug
interactions. The research was conducted as part of the installation research project of the Croatian Science Foundation
"Pharmacogenomics in Prediction of Cardiovascular Drugs Adverse Reaction (PGx CardioDrug)". Genotyping of
variants was performed by real-time PCR using TaqMan® SNP Assays on ABI 7500 Real-Time PCR System.
Furthermore, for each patient, a clinically relevant data such as age, gender, dose, BMI, comorbidities, eGFR, serum
creatinine level and concomitant therapy was collected from their clinical documentation. 96 patients treated with
dabigatran participated in this study (41 women and 55 men, age 21-93). Six patients were excluded from statistical
analysis due to exclusion criteria. Bleeding occurred in twenty-seven patients (30%), whereas only eight patients
experienced a thromboembolic event (8,9%). CES1 rs2244613 polymorphism was excluded from statistical analysis due
to deviation from Hardy-Weinberg equilibrium. We found no significant association between CES1 rs8192935 variant
and risk of bleeding or thromboembolic events which is consistent with previous studies. A statistically significant
association of female gender, lower dose (110 mg), impaired renal function and interactions between dabigatran and
drugs that potentially reduce its effectiveness with the risk of bleeding was found. Since the main limitation of this study
is a small sample size, in the future it is necessary to conduct a more comprehensive study on a larger sample to confirm
the association of CES1 variants with clinical outcomes, as well as the potential clinical benefit of genetic testing with
the aim of identifying patients at higher risk and optimizing therapy.

• Keywords: CES1; dabigatran; farmakogenetika; interakcije lijek-lijek; neželjeni klinički ishodi; polimorfizmi jednog nukleotida; SNP
• Keywords (english): CES1; dabigatran; pharmacogenetics; drug-drug interactions; adverse clinical outcomes; single nucleotide polymorphisms; SNP
• Language: croatian
• URN:NBN: urn:nbn:hr:163:191751
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije (magistar/magistra medicinske biokemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-01-18 14:16:29