20.500.12556/RUL-166068 Uspešnost imunoselekcije plazmocitomskih celic pri diseminiranem plazmocitomu Immunoselection efficiency of plasmacytoma cells in multiple myeloma Diseminirani plazmocitom (DP) je maligna bolezen plazmatk, za katero je značilno nenadzorovano razraščanje plazmocitomskih celic v kostnem mozgu (KM), kar zavira normalno hematopoezo in lahko povzroči resne zaplete. Najpogosteje se bolezen razvije iz monoklonske imunoglobulinemije neopredeljenega pomena (MINP), ki je običajno asimptomatska. Diagnostika DP temelji na odvzemu vzorca KM s punkcijo ali biopsijo, na katerem se izvajajo citomorfološki pregled, imunofenotipizacija z metodo pretočne citometrije in analiza kromosomskih nepravilnosti s tehniko fluorescenčne in situ hibridizacije (FISH). FISH omogoča odkrivanje klinično pomembnih kromosomskih nepravilnosti, ki vplivajo na prognozo in izbiro zdravljenja. Kljub temu se pri njeni izvedbi pogosto soočamo z izzivi, kot so neenakomerna razporeditev plazmatk v KM, hemodilucija vzorca s periferno krvjo in določene podvrste bolezni brez infiltracije KM. Analizirali smo uspešnost izvajanja preiskave FISH pri bolnikih s sumom na DP in preučili povezanost rezultatov citomorfološkega pregleda, pretočne citometrije in FISH. Prav tako smo primerjali tudi učinkovitost ročne in avtomatske imunoselekcije plazmatk. V raziskavo je bilo vključenih 811 bolnikov, obravnavanih v Specializiranem hematološkem laboratoriju (SHL) med leti 2019 in 2023. Rezultati so pokazali, da je bila imunoselekcija uspešna pri 83 % bolnikov, pri preostalih 17 % pa preiskava ni bila uspešno izvedena zaradi premajhnega deleža plazmatk v vzorcu KM. Pri bolnikih s potrjeno diagnozo DP je bila uspešnost imunoselekcije 98 %, kar pomeni, da le pri 2 % bolnikov s potrjenim DP preiskave FISH ni bilo mogoče izvesti. Pri 15 % vzorcev z uporabo nabora DNA-sond nismo odkrili kromosomskih preureditev. Pri bolnikih z dejansko potrjenim DP pa je bil delež negativnih rezultatov FISH bistveno nižji (2 %). Primerjava ročne in avtomatske imunoselekcije plazmatk ni pokazala statistično pomembnih razlik v uspešnosti izvedenih preiskav FISH ali številu uporabljenih DNA-sond. Prav tako smo potrdili, da sta pretočna citometrija in citomorfološka ocena pomembna napovedna dejavnika uspešnosti preiskave FISH. Rezultati citomorfološkega pregleda KM so pomemben napovedni dejavnik uspešnosti imunoselekcije, saj so imeli bolniki z več kot 10 % infiltracijo plazmatk v KM bistveno večjo verjetnost, da bo preiskava FISH uspešno izvedena ter da bo rezultat preiskave pozitiven. Rezultati so pokazali tudi, da je delež plazmatk, pridobljen s citomorfološko analizo, pomembno povezan s številom uporabljenih DNA-sond. Sklepamo, da je preiskava FISH ključnega pomena za diagnostiko in obravnavo bolnikov z DP, saj omogoča natančno odkrivanje kromosomskih nepravilnosti. Čeprav je uspešnost te metode visoka, njeno izvedbo pri nekaterih bolnikih otežuje nizka koncentracija plazmatk v vzorcu KM. Multiple myeloma (MM) is a malignant disease of plasma cells characterized by the uncontrolled proliferation of myeloma cells in the bone marrow (BM), which disrupts normal hematopoiesis and can lead to severe complications. The disease most commonly develops from monoclonal gammopathy of undetermined significance (MGUS), which is usually asymptomatic. The diagnosis of MM relies on the collection of BM samples via aspiration or biopsy, followed by cytomorphological examination, immunophenotyping by flow cytometry, and chromosomal abnormality analysis using fluorescence in situ hybridization (FISH). FISH enables the detection of clinically significant chromosomal abnormalities that influence prognosis and therapeutic decision-making. However, its application often faces challenges, such as the uneven distribution of plasma cells in BM, dilution of the sample with peripheral blood (hemodilution), and certain disease subtypes that do not infiltrate the BM. We analyzed the success of FISH testing in patients with suspected MM and examined the correlation between the results of cytomorphological examination, flow cytometry, and FISH. Additionally, we compared the efficiency of manual and automated plasma cell immunoselection. The study included 811 patients treated at the Specialized Hematology Laboratory (SHL) between 2019 and 2023. The results showed that immunoselection was successful in 83% of patients, while in the remaining 17%, FISH testing could not be performed due to an insufficient number of plasma cells in the BM sample. Among patients with confirmed MM, the success rate of immunoselection was 98%, indicating that FISH testing was not feasible in only 2% of cases with confirmed MM. In 15% of samples analyzed with the DNA probe panel, no chromosomal rearrangements were detected. However, among patients with confirmed MM, the proportion of negative FISH results was significantly lower (2%). The comparison of manual and automated plasma cell immunoselection revealed no statistically significant differences in the success rate of FISH testing or the number of DNA probes used. We also confirmed that both flow cytometry and cytomorphological assessment are significant predictors of FISH test outcomes. The results of cytomorphological BM examination were identified as a critical predictor of immunoselection success, as patients with more than 10% plasma cell infiltration in the BM had a significantly higher likelihood of successful FISH testing and positive results. Furthermore, the proportion of plasma cells obtained through cytomorphological analysis was significantly correlated with the number of DNA probes used. In conclusion, FISH testing is essential for the diagnosis and management of patients with MM, as it enables the precise detection of chromosomal abnormalities. Although the overall success of this method is high, its application in certain patients is hindered by the low plasma cell concentration in BM samples. FISH diseminirani plazmocitom imunoselekcija plazmocitomske celice kostni mozeg FISH multiple myeloma immunoselection plasmacytoma cells bone marrow true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2024-12-19 08:45:09 2024-12-19 08:45:13 2024-12-20 04:00:45 0000-00-00 00:00:00 2024 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 114485 26737.pdf 26737.pdf 1 BA8E493C3AAFB5A9CA6B820C5AA76A15 9cce206c606ee65f22cedecbc1d676981c8f71b96adf33f1fb570849f0d71487 9de90ec9-bddc-11ef-b232-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=197444 Fakulteta za farmacijo 0 0 0