Immunoglobulin genes and T-cell receptors as molecular markers in children with acute lymphoblastic leukaemia

Uvod. Akutna limfoblastna leukemija (ALL) je maligna klonalna bolest i jedna od najčešćih neoplazmi dečjeg uzrasta. Primenom savremenih protokola postižu se visok stepen remisije i dugogodišnja stopa preživljavanja. Uvođenjem metoda molekularne genetike omogućeni su submikroskopska klasifikacija i praćenje minimalne rezidualne bolesti (MRB), odgovorne za nastanak recidiva. Cilj rada. Cilj rada je bilo ispitivanje učestalosti rearanžmana genskih lokusa za teške lance imunoglobulina (IgH) i T-ćelijski receptor (TCR) i njihova korelacija s kliničkim parametrima. Metode rada. Ispitano je 41 dete obolelo od ALL kojima je na dijagnozi rađena molekularna detekcija rearanžmana genskih lokusa za IgH i TCR metodom lančanog umnožavanja DNK (PCR). Posmatranje razvoja MRB je rađeno u indukcionoj fazi lečenja, kada se očekuje morfološki odgovor na terapiju. Rezultati. Rearanžman genskog lokusa za IgH zabeležen je kod 82,9%, a za TCR kod 56,1% ispitanika. Posle 33 dana indukcione terapije rearanžmani za IgH genski lokus su zabeleženi kod 39%, a za TCR kod 36,5% dece. Zaključak. Otkrivanje genetskih aberacija na molekularnom nivou i njihova korelacija sa standardnim prognostičkim parametrima ukazala je na značaj nove stratifikacije rizičnih grupa, koje zahtevaju promenu intenziteta hemioterapije. Praćenje MRB se pokazalo preciznim prediktivnim faktorom u nastanku recidiva bolesti. PR Projekat Ministarstva nauke Republike Srbije, br. 143051.Introduction. Acute lymphoblastic leukaemia (ALL) is a malignant clonal disease, one of the most common malignancies in childhood. Contemporary protocols ensure high remission rate and long term free survival. The ability of molecular genetic methods help to establish submicroscopic classification and minimal residual disease (MRD) follow up, in major percent responsible for relapse. Objective. The aim of the study was to detect the frequency of IgH and TCR gene rearrangements and their correlation with clinical parameters. Methods. Forty-one children with ALL were enrolled in the study group, with initial diagnosis of IgH and TCR gene rearrangements by polimerase chain reaction ( PCR). MRD follow-up was performed in induction phase when morphological remission was expected, and after intensive chemiotherapy. Results. In the study group IgH rearrangement was detected in 82.9% of children at the diagnosis, while TCR rearrangement was seen in 56.1%. On induction day 33, clonal IgH rearrangements persisted in 39% and TCR rearrangements in 36.5% of children. Conclusion. Molecular analysis of genetic alterations and their correlation with standard prognostic parameters show the importance of risk stratification revision which leads to new therapy intensification approach. MRD stands out as a precise predictive factor for the relapse of disease

Importance of pharmacogenetic markers in the methylenetetrahydrofolate reductase gene during methotrexate treatment in pediatric patients with acute lymphoblastic leukemia

Despite remarkable progress in survival of children with acute lymphoblastic leukemia (ALL) which has reached about 85%, early toxicity and relapse rate remain issues that need to to be resolved. Genetic variants are important factors influencing the metabolism of cytotoxic drugs in ALL treatment. Variants in genes coding for methotrexate (MTX)-metabolizing enzymes are under constant scientific interest due to their potential impact on drug toxicity and relapse rate. We investigated methylenetetrahydrofolate reductase (MTHFR) c.677C gt T and MTHFR c.1298A gt C variants as pharmacogenetic markers of MTX toxicity and predictors of relapse. The study enrolled 161 children with ALL, treated according to the current International Berlin-Frankfurt-Munster group (BFM) for diagnostics and treatment of leukemia and lymphoma protocols. Genotyping was performed using PCR-RFLP and allele-specific PCR assays. Our results revealed similar distributions of MTHFR c.677C gt T and MTHFR c.1298A gt C genotypes among 104 healthy individuals as compared to pediatric ALL patients. A lower incidence of early MTX toxicity was noted in the MTHFR c.677TT genotype (p=0.017), while MTHFR c.1298A gt C genotypes were not associated with MTX toxicity. Carriers of any MTHFR c.677C gt T and MTHFR c.1298A gt C genotypes did not experience decreased overall survival (OAS) or higher relapse rates. Genetic variants in the MTHFR gene are not involved in leukemogenesis in pediatric ALL. The presence of the MTHFR c.677TT genotype was recognized as a predictive factor for decreased MTX toxicity during the intensification phase of therapy. Neither MTHFR c.677C gt T nor MTHFR c.1298A gt C genotypes correlated with an increased number of toxic deaths or relapse rate. Our study emphasizes the importance of implementing pharmacogenetic markers in order to optimize pediatric ALL therapy

ACTIVITIES OF PROXIMAL TUBULE ENZYMES AND ALBUMIN CONCENTRATION IN URINE OF CHILDREN TREATED WITH METHOTREXATE

In order to study methotrexate nephrotoxicity, the activities of proximal tubule epithelial cell membrane enzymes: alanine aminopeptidase (AAP) and gamma-glutamyltransferase (GGT), as well as of lysosomal N-acetyl-beta-D-glucosaminidase (NAG) and urinary albumin concentrations were determined in 12-h-urine samples of 30 patients with lymphoblastomous leukemia. The patients were i.v. receiving 4 individual methotrexate doses of 2000 mg/m2 every 15 days followed by leucovorin as a protector. Control and methotrexate-treated group, each consisting of 30 examinees, included 4–10 years old children of both sexes.Statistically significant increase of AAP and GGT activities, expressed as U/mmol creatinine was observed after the first two (p < 0.05), as well as after the remaining two therapies (p < 0.01) in relation to the control. Enzymatic activities of these two enzymes decreased to control value before the second and the third methotrexate application, but they increased again after the third application and remained elevated up to the end of experiments. Significant increase of NAG activity expressed as U/mmol creatinine (p < 0.01), as well as urinary albumin levels (mg/mmol creatinine; p < 0.01) were registered after the third methotrexate therapy and this elevation of the same statistical significance of the differences remained stable till the end of the therapy. Based on these results it can be concluded that during the time period of two first applications nephrotoxic methotrexate action is reversible and at the level of proximal tubule epithelial cell membranes. During the last two applications impairment is irreversible and at the level of cell organelles and glomerular filtration.In order to study methotrexate nephrotoxicity, the activities of proximal tubule epithelial cell membrane enzymes: alanine aminopeptidase (AAP) and gamma-glutamyltransferase (GGT), as well as of lysosomal N-acetyl-beta-D-glucosaminidase (NAG) and urinary albumin concentrations were determined in 12-h-urine samples of 30 patients with lymphoblastomous leukemia. The patients were i.v. receiving 4 individual methotrexate doses of 2000 mg/m2 every 15 days followed by leucovorin as a protector. Control and methotrexate-treated group, each consisting of 30 examinees, included 4–10 years old children of both sexes.Statistically significant increase of AAP and GGT activities, expressed as U/mmol creatinine was observed after the first two (p < 0.05), as well as after the remaining two therapies (p < 0.01) in relation to the control. Enzymatic activities of these two enzymes decreased to control value before the second and the third methotrexate application, but they increased again after the third application and remained elevated up to the end of experiments. Significant increase of NAG activity expressed as U/mmol creatinine (p < 0.01), as well as urinary albumin levels (mg/mmol creatinine; p < 0.01) were registered after the third methotrexate therapy and this elevation of the same statistical significance of the differences remained stable till the end of the therapy. Based on these results it can be concluded that during the time period of two first applications nephrotoxic methotrexate action is reversible and at the level of proximal tubule epithelial cell membranes. During the last two applications impairment is irreversible and at the level of cell organelles and glomerular filtration

ALKALINE PHOSPHATASE ENZYME AND LACTATE DEHYDROGENASE ACTIVITY IN URINE OF PATIENTS TREATED WITH METHOTREXATE

In order study methotrexate nephrotoxicity, the activities of proximal tubule epithelial cell membrane enzymes: alkaline phosphatase (AP) and lactate dehydrogenase (LDH) were determined in 12-h-urine samples of 30 patients with lymphoblastomous leukemia. The patients were i.v. receiving 4 individual methotrexate doses of 2000 mg/m2 every 15 days followed by leucovorin as a protector. Control and methotrexate-treated group, each consisting of 30 examinees, included 4-10 years old children of both sexes. Statistically significant increase of AP and LDH activities, expressed as unuts/mmol creatinine was observed after the second therapy (p < 0.05) in relation to the control. Based on these results it can concluded that nephrotoxic methotrexate action is ireversible during the time period after the second applications at the level of proximal tubule epithelal cell.In order study methotrexate nephrotoxicity, the activities of proximal tubule epithelial cell membrane enzymes: alkaline phosphatase (AP) and lactate dehydrogenase (LDH) were determined in 12-h-urine samples of 30 patients with lymphoblastomous leukemia. The patients were i.v. receiving 4 individual methotrexate doses of 2000 mg/m2 every 15 days followed by leucovorin as a protector. Control and methotrexate-treated group, each consisting of 30 examinees, included 4-10 years old children of both sexes. Statistically significant increase of AP and LDH activities, expressed as unuts/mmol creatinine was observed after the second therapy (p < 0.05) in relation to the control. Based on these results it can concluded that nephrotoxic methotrexate action is ireversible during the time period after the second applications at the level of proximal tubule epithelal cel

The state of research into children with cancer across Europe : new policies for a new decade

Overcoming childhood cancers is critically dependent on the state of research. Understanding how, with whom and what the research community is doing with childhood cancers is essential for ensuring the evidence-based policies at national and European level to support children, their families and researchers. As part of the European Union funded EUROCANCERCOMS project to study and integrate cancer communications across Europe, we have carried out new research into the state of research in childhood cancers. We are very grateful for all the support we have received from colleagues in the European paediatric oncology community, and in particular from Edel Fitzgerald and Samira Essiaf from the SIOP Europe office. This report and the evidence-based policies that arise from it come at a important junction for Europe and its Member States. They provide a timely reminder that research into childhood cancers is critical and needs sustainable long-term support.peer-reviewe

SIDEROPENIJSKA ANEMIJA U ADOLESCENCIJI

Sideropenijska anemija je najčešći zdravstveni problem dojenčadi i male djece u cijelomsvijetu. Globalna prevalencija sideropenijske anemije prema Svjetskoj zdravstvenojorganizaciji iznosi 32,9% (1). Adolescencija je vrijeme povećanih potreba za željezomi adolescenti imaju često neprepoznatu i/ili kasno otkrivenu sideropenijsku anemiju.Kada se pojave simptomi, oni su uzrokovani prvenstveno anemijom i uključujuslabost, glavobolju, razdražljivost i različit stepen umora te brzo zamaranje u fizičkomnaporu. Osim toga, dugotrajni manjak željeza može narušiti kognitivnu funkciju, koncentracijui sposobnost učenja. Simptomi se razvijaju postupno i dugo se zanemaruju.Laboratorijske manifestacije nedostatka željeza javljaju se u nekoliko faza i karakterišeih progresivno nastajanje anemije. Faktori rizika za razvoj sideropenije a potom i sideropenijskeanemije su: povećanje mišićne i ukupne tjelesne mase te povećanje volumenakrvi, obilni menstrualni gubici krvi, pothranjenost, teški tjelesni napori, vegetarijanskaprehrana ali i pretilost i infekcija s Helicobacter pylori kao i hronične bolesti.Kriteriji za anemiju zavise od životne dobi i za adolescente iznose: djevojčice: 12 godinai starije: feritin <15 μgr/L i hemoglobin <12 g/dL; a za dječake: od 12 do 15 godina:feritin <15 μgr/L i hemoglobin <12 g/dL a za dječake koji su 15 godina i stariji: feritin<15 μgr /L i hemoglobin <13 g/dL

PRIMARNI POREMEĆAJI IMUNITETA

Primarni poremećaji imuniteta (primarne imunodeficijencije) spadaju ugrupu rijetkih oboljenja. Postoji više pokušaja da se na sveobuhvatan način sagleda dijagnostičkai laboratorijska evaluacija primarnih poremećaja imuniteta i predstavi ljekarimakoji se primarno ne bave imunologijom, a dolaze u kontakt sa ovakvim pacijentima.Evropsko društvo za imunodeficijencije i druge kolege napravili su više dijagnostičkihprotokola za primarne poremećaje imuniteta. Primarni poremećaji imunitetamogu biti prisutni u svim životnim dobima te je evaluacija značajna kako za pedijatretako i za ljekare koji liječe odrasle pacijente

PREDIKTIVNI FAKTORI AKUTNE TOKSIČNOSTI TERAPIJE U DJECE SA AKUTNOM LIMFOBLASTNOM LEUKEMIJOM

Savremenom terapijom akutne limfoblastne leukemije (ALL) koja podrazumijeva intenzifikaciju imodeliranje citostatske terapije, danas je postignut značaj napredak u liječenju ovih bolesnika. Mi smo analizirali mogućeprediktivne faktore akutne toksičnosti hemioterapije u toku liječenja ALL dječije dobi

HISTIOCITOZA LANGERHANSOVIH ĆELIJA - OD JEDNOSISTEMSKOG DO MULTISISTEMSKOG OBLIKA

Histiocitoza Langerhansovih ćelija je rijetko oboljenje nastalo usljed proliferacije antigen prezentujućihLangerhansovih dendriditičnih ćelija iz koštane srži koje infiltrišu različite organe. Bolest je nepredvidivog toka, od mogućespontane remisije pa do teških diseminovanih oblika sa oštećenjem vitalnih organa i letalnim ishodom uprkos primjenjenojterapiji.U radu je prikazan slučaj 22 mjeseca stare djevojčice kod koje su se od drugog mjeca života javile promjenepo koži sa tendencijom pogoršanja tokom vremena. U dobi od 22 mjeseca djevojčici je verifikovana tumorska masa izalijevog uha sa znacima diabetes insipidusa (žedjanje, poliurija, polidipsija). Dijagnoza LCH je postvaljena patohistološki naosnovu bioptata kože i tumorske mase lijeve temporalne kosti. Uprkos terapiji bolest se reaktivirala nakon godinu danaterapije zbog čega je primjenjena i sekundarna hemoterapija. Usljed razvijenog diabetes inspidusa uvedena je doživotnasupstituciona terapija antiduretskim hormonom.Izolovane kožne lezije koje ne reaguju na lokalnu terapiju i koje imaju tendenciju pogoršanja bi trebale pobuditisumnju na LCH. U ovim slučajevima dodatna ispitivanja i bipsija kože su mandatorni radi blagovremene potvrde dijagnozei adekvatnog liječenja. Uprkos terapiji djeca sa razvijenim multisistemskim oblikom bolesti mogu imati neadekvatanodgovor na terapiju

Immunoglobulin genes and T-cell receptors as molecular markers in children with acute lymphoblastic leukaemia

Introduction. Acute lymphoblastic leukaemia (ALL) is a malignant clonal disease, one of the most common malignancies in childhood. Contemporary protocols ensure high remission rate and long term free survival. The ability of molecular genetic methods help to establish submicroscopic classification and minimal residual disease (MRD) follow up, in major percent responsible for relapse. Objective. The aim of the study was to detect the frequency of IgH and TCR gene rearrangements and their correlation with clinical parameters. Methods. Forty-one children with ALL were enrolled in the study group, with initial diagnosis of IgH and TCR gene rearrangements by polimerase chain reaction ( PCR). MRD follow-up was performed in induction phase when morphological remission was expected, and after intensive chemiotherapy. Results. In the study group IgH rearrangement was detected in 82.9% of children at the diagnosis, while TCR rearrangement was seen in 56.1%. On induction day 33, clonal IgH rearrangements persisted in 39% and TCR rearrangements in 36.5% of children. Conclusion. Molecular analysis of genetic alterations and their correlation with standard prognostic parameters show the importance of risk stratification revision which leads to new therapy intensification approach. MRD stands out as a precise predictive factor for the relapse of disease