Immunophenotypic characterization of acute myeloid leukemia in adults and its role in the diagnosis,monitoring, and prognosis of the disease

U tezi su ispitivani savremeni aspekti primene imunofenotipizacije multiparametarskom protočnom citofluorimetrijom (IMPC) u dijagnostici i praćenju AML. Ispitivanjem je obuhvaćeno 320 odraslih bolesnika sa de novo AML, od kojih je 294 uključeno u retrospektivno ispitivanje (dijagnoza, klasifikacija i prognoza), dok je ostalih 26 bolesnika bilo uključeno u prospektivno ispitivanje (praćenje minimalne rezidualne bolesti - MRB). Bolesnici su klasifikovani kao AML-neklasifikovana (48%), AML sa rekurentnim genetskim poremećajima (37,4%) i AML sa znacima mijelodisplazije (14,6%). IPCM omogućava postavljanje dg AML analizom ks kod 98% bolesnika. Ispitivanjem 44 različita hLDM utvrđeno je da je njihova ekspresija na blastima deregulisana, o čemu govore aberacije u njihovoj ekspresiji kod svakog bolesnika. Heterogen ćelijski sastav populacije leukemijskih ćelija (leukemijski blasti i prekursori) utvrđen je kod 55% bolesnika sa AML. Imunološka i citomorfološka klasfikacija AML su saglasne kod 73% bolesnika, odnosno imunološka i SZO klasifikacija kod 68% bolesnika. Ispitivanje MRB sprovedeno je primenom jedne (42%) ili dve (58%) kombinacije IFSL po bolesniku. Primenom IMPC, pokazana je visoka učestalost MRB kod naših bolesnika sa AML posle lečenja indukcionom (69%) odnosno konsolidacionom terapijom (50%). Nivo MRB u ks bolesnika ≥0,1% NĆ posle indukcione terapije, svrstava bolesnike u grupu visokog rizika za razvoj relapsa bolesti. Ispitivanje prognostičkog značaja hLDM kod bolesnika sa AML, pokazalo je značajnu vezu između rane monocitne diferencijacije leukemijskih ćelija, ekspresije CD22 molekula i pojave rane smrti i (p<0,05), odnosno niže incidence kompletne remisije (p<0,05)...In the thesis are examined modern aspects of application immunophenotyping and multiparameter flow cytometry (IMPC) in the diagnosis and monitoring of AML. The study included 320 adult patients with de novo AML, of which 294 included in the retrospective study (diagnosis, classification and prognosis), while the other 26 patients were included in a prospective study (minimal residual disease - MRD). Patients were classified as AML-unclassified (48%), AML with recurrent genetic abnormalities (37.4%) and AML with signs of myelodysplasia (14.6%). IPCM allows setting dg AML by analysis of bone marrow (bm) in 98% of patients. By examining of 44 different HLDM, it was found that its expression is deregulated on the blasts, whereas at least one type of immunophenotypic aberrations was found per patient. Heterogeneous cellular composition of the population of leukemic cells (leukemic blasts and precursors) was found in 55% of patients with AML. Immunological and cytomorphological classification of AML agree with 73% of patients, respectively immunological and WHO classification in 68% of patients. MRD trial was conducted by one (42%) or two (58%) combination of IFSL per patient. By applying the IMPC, a high incidence of MRD was detected in our patients with AML, after induction (69%) and/or consolidation therapy (50%). The level of MRD in bm of patients ≥0,1% NC after induction therapy, classified patients in the high risk group for the development of relapse. The prognostic significance of HLDM in patients with AML, showed a significant association between early monocytic differentiation of leukemia cells, the expression of CD22 molecule and the appearance of early death (p<0.05), and lower incidence of complete remission (p<0.05)..

The emergence of non-secretory multiple myeloma during the non-cytotoxic treatment of essential thrombocythemia: A case report

Introduction. The emergence of multiple myeloma as a second malignancy in patients with essential thrombocythemia is extremely rare. Several cases have been published so far, pointing out the impact of a cytotoxic effect during treatment of essential thrombocythemia on the development of multiple myeloma. Case presentation. We report the case of a 52-year-old Caucasian man who presented to our hospital because of leukocytosis, a slightly decreased hemoglobin level and thrombocytosis. After a complete hematological work-up, essential thrombocythemia was diagnosed. The patient was included in a multicenter clinical study, treated with anagrelide and his platelet counts were maintained in the normal range for more than 3 years. A sudden drop in his hemoglobin level with normal leukocyte and platelet count occurred at the same time as a back pain. Magnetic resonance imaging of his spine revealed the existence of a pathological fracture of Th4, the collapse of the upper edge of Th7 and osteolytic lesions of multiple thoracic vertebrae. Repeated hematological examinations, including bone biopsy with immunohistochemistry, disclosed diagnosis of multiple myeloma of the non-secretory type. Conclusions: To the best of our knowledge this is the first published case in which multiple myeloma developed during the treatment of essential thrombocythemia with the non-cytotoxic drug anagrelide. Our attempts to find a common origin for the coexistence of multiple myeloma and essential thrombocythemia have not confirmed the genetic basis of their appearance. Further studies are needed to determine the biological impact of this coexistence

Immunophenotypic characterization of acute myeloid leukemia in adults and its role in the diagnosis,monitoring, and prognosis of the disease

U tezi su ispitivani savremeni aspekti primene imunofenotipizacije multiparametarskom protočnom citofluorimetrijom (IMPC) u dijagnostici i praćenju AML. Ispitivanjem je obuhvaćeno 320 odraslih bolesnika sa de novo AML, od kojih je 294 uključeno u retrospektivno ispitivanje (dijagnoza, klasifikacija i prognoza), dok je ostalih 26 bolesnika bilo uključeno u prospektivno ispitivanje (praćenje minimalne rezidualne bolesti - MRB). Bolesnici su klasifikovani kao AML-neklasifikovana (48%), AML sa rekurentnim genetskim poremećajima (37,4%) i AML sa znacima mijelodisplazije (14,6%). IPCM omogućava postavljanje dg AML analizom ks kod 98% bolesnika. Ispitivanjem 44 različita hLDM utvrđeno je da je njihova ekspresija na blastima deregulisana, o čemu govore aberacije u njihovoj ekspresiji kod svakog bolesnika. Heterogen ćelijski sastav populacije leukemijskih ćelija (leukemijski blasti i prekursori) utvrđen je kod 55% bolesnika sa AML. Imunološka i citomorfološka klasfikacija AML su saglasne kod 73% bolesnika, odnosno imunološka i SZO klasifikacija kod 68% bolesnika. Ispitivanje MRB sprovedeno je primenom jedne (42%) ili dve (58%) kombinacije IFSL po bolesniku. Primenom IMPC, pokazana je visoka učestalost MRB kod naših bolesnika sa AML posle lečenja indukcionom (69%) odnosno konsolidacionom terapijom (50%). Nivo MRB u ks bolesnika ≥0,1% NĆ posle indukcione terapije, svrstava bolesnike u grupu visokog rizika za razvoj relapsa bolesti. Ispitivanje prognostičkog značaja hLDM kod bolesnika sa AML, pokazalo je značajnu vezu između rane monocitne diferencijacije leukemijskih ćelija, ekspresije CD22 molekula i pojave rane smrti i (p<0,05), odnosno niže incidence kompletne remisije (p<0,05)...In the thesis are examined modern aspects of application immunophenotyping and multiparameter flow cytometry (IMPC) in the diagnosis and monitoring of AML. The study included 320 adult patients with de novo AML, of which 294 included in the retrospective study (diagnosis, classification and prognosis), while the other 26 patients were included in a prospective study (minimal residual disease - MRD). Patients were classified as AML-unclassified (48%), AML with recurrent genetic abnormalities (37.4%) and AML with signs of myelodysplasia (14.6%). IPCM allows setting dg AML by analysis of bone marrow (bm) in 98% of patients. By examining of 44 different HLDM, it was found that its expression is deregulated on the blasts, whereas at least one type of immunophenotypic aberrations was found per patient. Heterogeneous cellular composition of the population of leukemic cells (leukemic blasts and precursors) was found in 55% of patients with AML. Immunological and cytomorphological classification of AML agree with 73% of patients, respectively immunological and WHO classification in 68% of patients. MRD trial was conducted by one (42%) or two (58%) combination of IFSL per patient. By applying the IMPC, a high incidence of MRD was detected in our patients with AML, after induction (69%) and/or consolidation therapy (50%). The level of MRD in bm of patients ≥0,1% NC after induction therapy, classified patients in the high risk group for the development of relapse. The prognostic significance of HLDM in patients with AML, showed a significant association between early monocytic differentiation of leukemia cells, the expression of CD22 molecule and the appearance of early death (p<0.05), and lower incidence of complete remission (p<0.05)..