Važnost određivanja IDH1 i IDH2 mutacija u bolesnika s dijagnozom akutne mijeloične leukemije

master's thesis

• Title (english): Importance of detecting IDH1 and IDH2 mutations in patients with acute myeloid leukemia diagnosis
• Author: Marija Vasiljević
• Mentor: Dunja Rogić (mentor)
• Committee member: Dunja Rogić (predsjednik povjerenstva)
• Committee member: Ivana Lapić (član povjerenstva)
• Committee member: Željka Vogrinc (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2024-11-05, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Akutna mijeloična leukemija visoko je heterogena hematološka zloćudna bolest, koju karakterizira abnormalna
proliferacija nezrelih stanica mijeloidne loze. AML je bolest starije životne dobi s medijanom od 67 godina. U pozadini
AML-a stoje broje genetske promjene pa najnovije smjernice za stratifikaciju rizika i postavljanje dijagnoze ističu
važnost određivanja citogenetskih i genskih promjena. Iako je došlo do znatnog napretka u dijagnostici i razumijevanju
patofiologije AML-a, osnovna terapija (intenzivna kemoterapija) ostaje ista dok je petogodišnje ukupno preživljavanje
i dalje nisko posebno kod recidiva, starijih osoba, sekundarnog i rezistentnog AML. Odobrene spasonosne terapije
postoje samo za one s FLT3 te IDH1/2 mutacijama. Terapija IDH inhibitorima nije se pokazala učinkovitijom od
standardnih terapija kod novodijagnosticiranog AML-a, ali kod bolesnika s R/R AML-om omogućuje postizanje CR u
oko 20 % slučajeva dok stopa ukupnog odgovora uključujući i hematološko poboljšanje iznosi oko 40 %. Zbog toga je
rana detekcija ovih mutacija postala bitna. ELN preporuča da se genska analiza na prisutnost IDH1/2 mutacija provede
unutar 3 do 5 dana od zaprimanja uzoraka. Prema tome, cilj ovog rada bio je uspostaviti dijagnostički postupak za
dokazivanje najčešćih mutacija u genima IDH1 i IDH2 u rutinskoj laboratorijskoj dijagnostici. Odabran je brzi i osjetljivi
komercijalni test, kojim je analizirano 40 uzoraka izdvojene DNA bolesnika s dijagnozom AML-a kojima je prethodno
dokazana NPM1 mutacija. Dokazane su ukupno 22 mutacije – 12 u IDH1 genu (7 na kodonu R132 i 5 varijanti G105G)
i 10 u IDH2 genu (7 na kodonu R140 i 3 na kodonu R172). Dio rezultata potvrđen je metodom Sangerovog sekvenciranja
uz potpuno slaganje. Utvrđena je očekivana učestalost IDH mutacija u skupini bolesnika s NPM1 mutiranim AML-om
te test zadovoljava temeljne kriterije za uporabu u rutinskoj laboratorijskoj dijagnostici.

Abstract (english)

Acute myeloid leukemia (AML) is a highly heterogeneous hematologic malignancy marked by abnormal proliferation
of myeloid progenitors, primarily affecting elderly patients, with a median age of 67 years. Given the numerous genetic
abnormalities in AML, current guidelines for risk stratification and diagnosis underscore the importance of identifying
cytogenetic and genetic alterations. Although significant advances have been made in AML diagnostics and
understanding AML's pathophysiology, intensive chemotherapy remains the standard treatment. Still, 5-year overall
survival remains low, particularly for the elderly, relapsed, secondary, and resistant AML. Currently, approved targeted
therapies exist only for patients with FLT3 and IDH1/2 mutations. While IDH inhibitors have not demonstrated superior
efficacy to standard therapy in newly diagnosed AML, in relapsed/refractory AML, they enable complete remission in
about 20% of cases, with an overall response rate of approximately 40% when including hematologic improvements.
Thus, early detection of these mutations is critical. The European LeukemiaNet (ELN) recommends that genetic analysis
for IDH1/2 mutations be conducted within 3 to 5 days of sample collection. This thesis aimed to establish a diagnostic
protocol for the routine laboratory detection of IDH1 and IDH2 mutations. A fast, sensitive commercial test was
employed to analyze DNA from 40 AML patients with a previously confirmed NPM1 mutation. 22 mutations were
detected—12 in the IDH1 gene (7 at codon R132 and 5 G105G variants) and 10 in the IDH2 gene (7 at codon R140 and
3 at codon R172). Sanger sequencing confirmed a subset of results with full concordance. The study confirmed the
expected frequency of IDH mutations among NPM1-mutated AML patients and demonstrated that the chosen test meets
essential criteria for use in routine laboratory diagnostics.

• Keywords: akutna mijeloična leukemija; AML; IDH1; IDH2; mutacija; qPCR; rutinska dijagnostika
• Keywords (english): acute myeloid leukemia; AML; IDH1; IDH2; mutation; qPCR; Sanger sequencing; routine diagnostics
• Language: croatian
• URN:NBN: urn:nbn:hr:163:873380
• Study programme: Title: Medical biochemistry - integrated undergraduate and graduate - university study; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: sveučilišni magistar medicinske biokemije i laboratorijske medicine (sveučilišni magistar medicinske biokemije i laboratorijske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2025-02-05 12:11:17