Mutacija gena BRAF V600E u leukemiji vlasastih stanica

master's thesis

• Title (english): BRAF V600E gene mutations in hairy cell leukemia
• Author: Nikolina Žuljević
• Mentor: Rajko Kušec (mentor)
• Committee member: Vlatko Pejša (predsjednik povjerenstva)
• Committee member: Ljiljana Šerman (član povjerenstva)
• Committee member: Rajko Kušec (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; (Department of Internal Medicine); Zagreb
• Defense date and country: 2018-07-13, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Internal Medicine

Abstract

Leukemija vlasastih stanica ili triholeukemija je rijetka, limfoproliferativna bolest s malobrojnim cirkulirajućim tumorskim stanicama u perifernoj krvi. Trihociti su stanice čiji su citoplazmatski izdanci nalik vlasima te karakteriziraju ovu bolest. TL je kronična tijeka i sporo progresivna, te dobro reagira na terapiju purinskim analozima koji su standard u liječenju. Bolest uglavnom zahvaća slezenu, a limfni čvorovi su pošteđeni. TL je povezana s mutacijom BRAF gena i to s najčešćom mutacijom gena, BRAF V600E. Ta mutacija dovodi do konstitutivne aktivnosti RAS-RAF-MEK-ERK signalnog puta i do proliferacije stanica. BRAF mutacije do sad su povezane s brojnim malignomima, a najčešće s melanomom. Otkriće povezanosti triholeukemije s BRAF mutacijom donosi mogućnosti za novu terapiju BRAF inhibitorima, vemurafenibom i dabrafenibom, te praćenje bolesti uz pomoć RT-PCR metode. U ovom istraživanju analizirano je 15 ispitanika dijagnosticiranih s TL-om u KB Dubrava, u Zagrebu. Korišteni su uzorci periferne krvi i koštane srţi uklopljene u parafin. U istraživanju se utvrđivalo postojanje BRAF V600E mutacije u oboljelih te ocjenjivala uloga alel specifičnog RT-PCR-a u praćenju bolesti i otkrivanju minimalne ostatne bolesti. Zaključeno je da je RT-PCR metoda dovoljno osjetljiva za otkriće mutacije u aktivnoj bolesti te relapsu, a slabije osjetljiva u remisiji bolesti te kod minimalno aktivne bolesti. Povećan postotak limfocita u perifernoj krvi u ispitanika se pokazao kao dobar prediktor utvrđivanja BRAF V600E mutacije.

Abstract (english)

Hairy cell leukemia, also called trycholeukemia, is a rare lymphoproliferative disorder with few circulating tumor cells in the peripheral blood. Hairy cells, characteristic of this disorder, are cells that have hair-like cytoplasmic projections. HCL has a chronic course. It is slowly progressive and has a good response to the standard treatment with purine analogues. Usually it affects the spleen, and lymph nodes are spared. HCL is associated with the BRAF gene mutation, more precisely with the most common, BRAF V600E mutation. That mutation leads to the constitutive activity of the RAS-RAF-MEK-ERK signal pathway and cell proliferation. BRAF mutations are related to many malignomas, but most commonly to melanoma. The discovery of the association between the HCL and the BRAF mutation lead to new possibilites in the treatment of the disorder, such as BRAF inhibitors, vemurafenib, and dabrafenib, as well as the possibility to follow up the disease with the RT-PCR method. In our study, 15 patients diagnosed with HCL were analyzed in the Clinical Hospital Dubrava in Zagreb. We used peripheral blood samples and samples of bone marrow embedded in paraffin. In this study we tried to detect the BRAF V600E mutation in patients diagnosed with HCL, and we also evaluated the allele-specific RT-PCR in the follow up of the disease and in the detection of the minimal residual disease. We concluded that the RT-PCR method is sensitive enough to detect the mutation in the active disease and in the relapse, but is less sensitive in the remission and in the minimal residual disease. A high percentage of lymphocytes in the peripheral blood has proved to be a good predictor of the BRAF V600E mutation.

• Keywords: triholeukemija; leukemija vlasastih stanica; BRAF; BRAF V600E; RAS-RAF-MEK-ERK; vemurafenib; alel specifični RT-PCR
• Keywords (english): hairy cell leukemia; trycholeukemia; BRAF; BRAF V600E; RAS-RAF-MEK-ERK; vemurafenib; allele-specific RT-PCR
• Language: croatian
• URN:NBN: urn:nbn:hr:105:440667
• Study programme: Title: Medicine; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: doktor/doktorica medicine (doktor/doktorica medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2019-01-10 10:45:58