Farmakogenetički markeri odgovora na terapiju tiopurinskim lekovima, metotreksatom i vinkristinom kod dece sa akutnom limfoblastnom leukemijom

Abstract

Akutna limfoblastna leukemija (ALL) je najčešći malignitet dečjeg doba. Napredak u lečenju usmeren je ka smanjenju pojave neželjenih efekata terapije, primenom principa farmakogenetike. U ovoj studiji istraživanje je bilo usmereno ka otkrivanju farmakogenetičkih i farmakotranskriptomskih markera odgovora na lekove koji su okosnica terapijskih protokola lečenja dece obolele od ALL (tiopurinski lekovi, metotreksat i vinkristin). Analizirani su uzorci krvi i koštane srži 139 dece sa ALL. Varijante u ispitivanim genima detektovane su metodom PCR-a i Sangerovim sekvenciranjem. Nivo ekspresije NUDT15 u mononuklearnim ćelijama određen je real-time PCR-om. Korelacija farmakomarkera sa kliničkim parametrima vršena je statističkim testovima. Odgovor na ispitivane lekove procenjivan je u retrospektivnim kliničkim studijama. Pokazano je da nivo ekspresije NUDT15 na prezentaciji bolesti ne korelira sa pojavom neželjenih efekata terapije tiopurinskim lekovima. U ispitivanoj grupi detektovano je 5 varijanti u genu NUDT15 koje nemaju patogeni efekat. Varijanta rs4149056 u genu SLCO1B1 je izučavana kao marker farmakokinetike i toksičnosti metotreksata. U ovoj studiji nije potvrđen njen klinički značaj. Analiziran je farmakogenetički potencijal varijanti u genima CYP3A5, CEP72, ACTG1, MIR3117 i MIR4481 kao markera odgovora na terapiju vinkristinom. Nije potvrđena korelacija ovih varijanti sa nastankom perifernih neuropatija kod dece sa ALL. Razvijen je predikcioni model za hepatotoksičnost izazvanu metotreksatom, zasnovan na poligenskom skoru rizika. Ova studija je doprinela znanjima koja vode ka personalizaciji lečenja dece obolele od ALL.

Acute lymphoblastic leukemia (ALL) is the most common childhood malignancy. Treatment progress is aimed at reducing the occurrence of side effects of therapy, by applying the principles of pharmacogenetics. The aim of this study was to discover pharmacogenetic and pharmacotranscriptomic markers of response to drugs that are the backbone of therapeutic protocols for the treatment of children with ALL (thiopurine drugs, methotrexate and vincristine). Blood and bone marrow samples of 139 children with ALL were analysed. Variants in the tested genes were detected by PCR and Sanger sequencing. The expression level of NUDT15 in mononuclear cells was determined by real-time PCR. The correlation of pharmacomarkers with clinical parameters was performed using statistical tests. The response to the investigated drugs was evaluated in retrospective clinical studies. It was shown that the level of NUDT15 expression at the presentation of the disease does not correlate with the occurrence of side effects of thiopurine drug therapy. In this study 5 variants in the NUDT15 gene were detected, which do not have a pathogenic effect. The rs4149056 variant in the SLCO1B1 gene was assessed as a marker of methotrexate pharmacokinetics and toxicity. Its clinical significance was not confirmed in this study. The pharmacogenetic potential of variants in CYP3A5, CEP72, ACTG1, MIR3117 and MIR4481 genes as markers of response to vincristine therapy was analysed. The correlation of these variants with the onset of peripheral neuropathies in children with ALL has not been confirmed. A predictive model for methotrexate-induced hepatotoxicity based on the polygenic risk score was developed. This study contributed to the knowledge that leads to the personalization of the treatment of children with ALL.