Povezanost povrata bolesti i gubitka nepodudarnog haplotipa HLA kod haploidentične transplantacije krvotvornih matičnih stanica

master's thesis

• Title (english): Disease relapse and loss of mismatched HLA haplotype in haploidentical stem cell transplantation
• Author: Sara Bralić
• Mentor: Zorana Grubić (mentor)
• Committee member: Zorana Grubić (predsjednik povjerenstva)
• Committee member: Ana Galov (član povjerenstva)
• Committee member: Jasna Lajtner (član povjerenstva)
• Committee member: Petar Kružić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2019-12-12, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Haploidentična transplantacija krvotvornih matičnih stanica (TKMS) je metoda liječenja za bolesnike koji nemaju genotipski HLA identičnog davatelja u obitelji ili fenotipski HLA podudarnog nesrodnog davatelja. Cilj rada bio je istražiti učestalost gubitka primateljevog nepodudarnog haplotipa HLA među bolesnicima s povratom bolesti nakon haploidentične TKMS primjenom metode lančane reakcije polimerazom i početnicama specifičnih sekvenci (PCR-SSP). Bolesnicima s relapsom bolesti važno je utvrditi radi li se o klasičnom povratu bolesti ili relapsu s gubitkom nepodudarnog haplotipa HLA zbog nastavka liječenja drugom transplantacijom. Istraživanje je obuhvatilo skupinu nesrodnih bolesnika (N=28) i njihovih HLA haploidentičnih srodnih davatelja. Svim bolesnicima praćen je status kimerizma, odnosno postotak primateljevih i davateljevih stanica nakon TKMS metodom lančane reakcije polimerazom u stvarnom vremenu (qPCR). Bolesnicima s povratom bolesti, kao i njihovim davateljima, određeni su geni lokusa HLA-A, -B, -C, -DRB1 i -DQB1 metodom PCR-SSP, te su im određeni informativni lokusi koji su dalje korišteni u praćenju gubitka nepodudarnog haplotipa HLA. Povrat bolesti utvrđen je kod devet (32,1%) bolesnika, od kojih je kod dva (22,2%) bolesnika utvrđen gubitak nepodudarnog haplotipa HLA. Određivanje gena HLA metodom PCR-SSP pokazalo se primjenjivim i u slučajevima malog udjela primateljevih stanica (2%) što govori u prilog osjetljivosti ove metode u analizama gubitka nepodudarnog haplotipa HLA.

Abstract (english)

Hematopoietic stem cell transplantation (HSCT) from a haploidentical donor is a curative option for patients who lack an HLA genotypically identical donor or HLA matched unrelated donor. The aim of this study was to analyze the frequency of the genomic loss of the recipient's mismatched HLA haplotype (HLA loss) among patients who relapsed after haploidentical HSCT using the polymerase chain reaction and sequence specific primers (PCR-SSP) method. In the group of patients who relapsed it is important to determine whether it was a normal disease relapse or HLA loss relapse. Unrelated patients (N=28) and their HLA haploidentical related donors were included in this study. In all patients the percentage of recipient and donor cells after HSCT was determined using the Real-Time PCR (qPCR) method. HLA-A, -B, -C, -DRB1 and -DQB1 genes were typed in the group of patients who relapsed and their donors using the PCR-SSP method. Informative loci were defined and were used for HLA loss analysis. Disease relapse was detected in nine (32,1%) patients, and two (22,2%) of them had HLA loss. The results of this analysis demonstrated that PCR-SSP method is very sensitive in HLA loss analysis because it is possible to detect HLA genes even in cases of the small percentage of recipient cells (2%).

• Keywords: haploidentična transplantacija; nepodudarni haplotip HLA; geni HLA
• Keywords (english): haploidentical transplantation; mismatched HLA haplotype; HLA genes
• Language: croatian
• URN:NBN: urn:nbn:hr:217:966370
• Study programme: Title: Experimental Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2021-12-12
• Created on: 2020-06-09 10:51:34