Influence of clonal b cell proliferation and cytokine gene polymorphisms on disease severity and occurence of monoclonal gammopathy in patients with Gaucher disease

Gošeova bolest je oboljenje lizozoma kod kojeg je korelacija između genotipa i fenotipske ekspresije bolesti slaba te se samim tim ni težina bolesti ne može pouzdano predvideti. Citopatološke promene u ovoj bolesti nisu izazvane samo nakupljanjem nerazgrađenog glikolipida, već i aktivacijom makrofaga, a koncentracije različitih citokina, važnih za proces inflamacije, su povišene u serumu ovih bolesnika. Takođe, nekoliko studija u literaturi navode dokaze o povećanoj incidenciji hematoloških komplikacija u bolesnika sa Gošeovom bolešću, uključujući monoklonsku i poliklonsku gamapatiju i hematološke malignitete, a posebno multipli mijelom. U kohorti naših bolesnika u Srbiji je ranije već opisana visoka incidencija rearanžmana gena za teški lanac imunoglobulina, što sugeriše da se aktivacija imunskog sistema može javiti relativno rano tokom bolesti. Predmet rada istraživanja ove studije je bila analiza uticaja klonskog rearanžmana gena za teški lanac imunoglobulina i/ili prisustva pojedinih alela gena koji kodiraju molekule važne u inflamaciji (TNFα, IL-10 i IL-6) na težinu kliničke slike obolelih od Gošeove bolesti. Materijal i metod: U ovu kohortnu studiju su bili uključeni pacijenti oboleli od Gošeove bolesti u Srbiji koji su u periodu od 2008 – 2016. godine lečeni na Klinici za hematologiju i Klinici za endokrinologiju, dijabetes i bolesti metabolizma Kliničkog centra Srbije, u Beogradu, kao i Klinici za hematologiju Kliničkog centra u Nišu. Kriterijumi za isključenje iz studije su bili: odbijanje pacijenta da učestvuje u studiji, smrt pacijenta ili emigracija u druge države. Kontrolna grupa za poređenje učestalosti ispitivanih polimorfnih genotipova za citokinske gene je bila selektovana iz baze DNK uzoraka Laboratorije za imunologiju Instituta za Mikrobiologiju i imunologiju. Svaki od ispitanika je detaljno obavešten o cilju studije i potpisao je dobrovoljan pristanak za učestvovanje u njoj. Iz uzoraka pune venske krvi je vršena izolacija DNK, a detekcija i analiza polimorfizama citokinskih gena TNF, IL10 i IL6 je vršena metodom Real-time PCR, u Laboratoriji za imunologiju Instituta za mikrobiologiju i imunologiju Medicinskog fakulteta Univerziteta u Beogradu...Gaucher disease is a lysosomal storage disease showing weak genotype-phenotype correlation, with poorly predictable disease severity. Cytopathological alterations are caused not only by the accumulation of the undegraded glycolipid substrate, but also by macrophage activation, displaying serum elevation of various cytokines with important influence to inflammation. Several studies in the literature showed increased incidence of hematological complications in Gaucher patients, including monoclonal and polyclonal gammopathies as well as hematological malignancies, especially multiple myeloma. We had already described a high incidence of immunoglobulin heavy chain gene rearrangements in a cohort of Serbian patients, suggesting that an early occurrence of immune system activation is possible early in the disease course. The object of this study was the analysis of clonal rearrangements of IGH genes and frequencies of alleles of cytokine genes important for inflammation (TNFα, IL-10 and IL-6) as well as their influence on disease severity in Gaucher patients. Material and methods: the cohort study included patients with Gaucher disease treated in Serbia since 2008 – 2016, at the Clinic of hematology and Clinic for endocrinology, diabetes and metabolic diseases of Clinical center of Serbia in Belgrade, as well as Clinic of hematology, Clinical Center of Niš. Exclusion criteria were: patient refusal to participate in the study, death of a patient and / or emmigration of a patient. Control group for frequency analysis of polymorphic genotypes of cytokine genes was selected out of the database of DNA samples acquired from Laboratory of immunology, Institute of microbiology and immunology. Every subject was thoroughly informed of the aim of the study and signed the informed consent in order to participate in it voluntarily. Venous blood samples were taken with subsequent DNA isolation permitting detection and analysis of cytokine gene polymorphisms (TNF, IL10 and IL6) using the method of Real-time PCR. This was performed in the Laboratory of immunology, Institute of microbiology and immunology, School of medicine, University of Belgrade..