Мантийноклеточная лимфома (МКЛ) – агрессивная и тяжело поддающаяся терапии лимфома из зрелых В-лимфоцитов, характеризующаяся транслокацией t(11;14) (q13;q32) и гиперэкспрессией циклина D1. У большинства пациентов заболевание отличается прогрессирующим клиническим течением, требующим незамедлительного начала противоопухолевой терапии, однако наряду с агрессивными вариантами выявляются индолентные МКЛ с длительной выживаемостью и возможностью применения тактики «наблюдай и жди». Такое различие обусловливается наличием вторичных цито- и молекулярно-цитогенетических аберраций, выявляющихся в дополнение к высокоспецифической t(11;14)(q13;q32), которые усиливают онкогенный потенциал циклина D1, вовлекают в лимфомогенез гены, регулирующие клеточный цикл, апоптоз, репарацию поврежденной ДНК, таким образом способствуя увеличению нестабильности генома и дальнейшей злокачественной трансформации опухолевого клона. В настоящее время особое внимание уделяется изучению прогностического влияния изменений кариотипа и аберраций отдельных генов, таких как MYC/8q24 и ТР53/17р13, на выживаемость пациентов с МКЛ. В нашем исследовании мы оценили генетический профиль 117 пациентов с МКЛ для выявления прогностически значимых цитогенетических и молекулярно-генетических маркеров и их взаимосвязи с продолжительностью общей и безрецидивной выживаемости пациентов. Mantle cell lymphoma (MCL) is an aggressive and difficult to treat lymphoma of mature B-lymphocytes characterized by t(11;14)(q13;q32) translocation and overexpression of cyclin D1. In most cases, the disease is manifested by progressive use of therapy, requiring the expense of starting anticancer therapy, however, in combination with aggressive variants, indolent MCL with long-term survival and the possibility of "watch and wait" tactics are detected. This difference is due to the presence of secondary cytogenetic and molecular cytogenetic aberrations, which are revealed in addition to the highly specific translocation t(11;14)(q13;q32), which enhance the oncogenic potential of cyclin D1, involve genes that regulate the cell cycle, apoptosis, and repair in lymphomagenesis. damaged DNA, thus contributing to an increase in genome instability and further malignant transformation of the tumor clone. Currently, special attention is paid to the study of the prognostic impact of changes in the karyotype and aberrations of individual genes, such as MYC/8q24 and TP53/17p13, on the survival of patients with MCL. In our study, we assessed the genetic profile of 117 patients with MCL to identify prognostic cytogenetic and molecular genetic markers and their relationship with the duration of overall and disease-free survival of patients.

Mantle cell lymphoma (MCL) is a type of peripheral B-cell non-Hodgkin’s lymphoma characterized by constitutive cyclin D1 overexpression leading to cell-cycle dysregulation and disruption of DNA damage repair. Apart from the typical translocation t(11;14)(q13;q32) and more rare variants, such as t(2;11)(p11;q13) and t(11;22)(q13;q11), a considerable number of patients quite often show secondary molecular and chromosomal aberrations underlying heterogeneity of the clinical course of MCL. Among a wide range of molecular genetic abnormalities, particular attention during the last years has been concentrated on studying the so-called double-hit MCL within a subgroup of patients with translocations involving CCND1 and MYC genes. Double-hit MCL is distinguished with rapid progression and tumor generalization at the time of diagnosis. Poor prognosis and low survival rates in most MCL patients call for the fastest possible diagnosis. Morphological and immunohistochemical as well as genetic methods (standard cytogenetic technique and fluorescence in situ hybridization) contribute to improving the quality of evidence-based diagnosis. The results of comprehensive diagnostic studies optimize prognosis assessment and treatment decision making in clinic.

Aim. To study the relationship of karyotype, JAK2, CALR, and MPL driver mutations and ASXL1 mutation status with the progression and prediction of primary myelofibrosis (PMF). Materials & Methods. The trial included 110 PMF patients (38 men and 72 women), median age was 59 years (range 18-82) with median follow-up after diagnosis of 2.6 years (range 0.1-23). The patients were examined for JAK2, CALR, MPL, and ASXL1 mutations. Restriction fragment length polymorphism technique was used for the analysis of V617F substitution in JAK2 and 515 codon mutation in MPL. CALR (exon 9) and ASXL1 (exon 12) mutation tests were performed using Sanger direct sequencing. In 48 (44 %) out of 110 patients bone marrow cell karyotype was determined. Clinical and hematological parameters and median overall survival (OS) of patients were analyzed with regard to detected genetic aberrations and combinations of them. Results. JAK2, CALR, MPL mutations were detected in 55 (50 %), 28 (25.5 %), and 7 (6.4 %) out of 110 patients, respectively. Triple negative (TN) status was identified in 20 (18.2 %) out of 110 examined patients. ASXL1 mutations were detected in 22 (20 %) out of 110 patients. Out of 48 patients in 32 (66.7 %) normal karyotype, in 3 (6.3 %) favorable karyotype, in 4 (8.3 %) intermediate-prognosis karyotype, and in 9 (18.7 %) unfavorable karyotype were detected. The comparison of clinical and hematological parameters showed a number of significant differences. JAK2-positive patients had a higher hemoglobin level (median 129 g/L; p = 0.021). TN was associated with a high IPSS risk (p = 0.011), low hemoglobin level (median 101 g/L; p = 0.006), drop in platelet count (median 266 x 109/L; p = 0.041), increased lymphocyte count (median 26.9 х 109/L; р = 0.001). The detection of terminating mutations in ASXL1 correlated with palpable enlarged spleen (р = 0.050), reduced platelet count (median 184 х 109/L; р = 0.016), leukocyte count > 25 х 109/L (р = 0.046), and blast count > 1 % (р < 0.001). Univariate regression analysis showed that terminating mutations in ASXL1 (hazard ratio [HR] 2.9; р = 0.018), unfavorable karyotype (HR 8.2; р < 0.001), and TN (ОР 8.1; р < 0.001) had prognostic value for OS. ASXL1 mutation was associated with significantly worse OS in TN patients. Median OS of ASXL1-negative patients without high-risk chromosomal aberrations was significantly longer than in patients with high-risk karyotype and/ or ASXL1 mutation. Conclusion. Several genetic defects in tumor cells are associated with phenotypic manifestations of PMF. Based on the results of cytogenetic analysis and mutation determination of JAK2, CALR, MPL, and ASXL1, patients can be classified in different “genetic” risk groups when PMF is diagnosed.

Aim. To analyze the effect on prognosis of mutations that are typical of acute myeloid leukemia (AML) patients. Materials & Methods. The study included 620 AML patients surveyed at Hematology Clinics of Saint Petersburg (Russia) and Charite Clinic (Berlin, Germany). G-banding of chromosomes was employed for cytogenetic testing. Aberration screening in DNMT3A, IDH1/2 genes was based on real-time polymerase chain reaction (PCR) with subsequent analysis of melting and sequencing profiles. Mutations in FLT3, NPM1 genes were revealed by PCR. Results. Mutations were identified in 343 (55.3 %) out of 620 patients. Significantly more often mutations were discovered in patients with normal karyotype (NK) (p = 0.001). FLT3-ITD mutation was associated with reduced medians of overall survival (OS) and disease-free (DFS) survival: 11.3 vs. 15.8 months with FLT3-ITD- (p = 0.005) and 10.0 vs. 13.3 months with FLT3-ITD+ (p = 0.009), respectively. The relation of FLT3-ITD allele burden to OS duration was also assessed. In the ITDlow/ITD- group the OS median was considerably longer than in the ITDhigh group (p = 0.028). In the group of patients with 1 mutation in NPM1 gene OS and DFS were much better in comparison with other patients (medians of 27.4 and 13.9 months, respectively, p = 0.040; 19.3 and 12.0 months, p = 0.049). Negative impact of mutations in DNM-T3A gene was noticed while assessing OS median: 12 (DN-MT3A+) and 15 months (DNMT3A-), respectively (p = 0.112). Mutations in IDH1 gene correlated with a better OS than in the group without mutations (p = 0.092). The rs11554137 polymorphism in IDH1 gene was associated with worse OS in the group of patients with NK (p = 0.186). In 144 patients various mutation combinations (from 2 to 5) were identified. It was demonstrated that mutations in FLT3 (FLT3-ITD), NPM1, DNMT3A, and IDH2 were identified significantly more often in combinations with other mutations (p = 0.001): NPM1+/ FLT3-ITD+ (20.8 %), NPM1+/FLT3-ITD+/DNMT3A+ (8.3 %), and FLT3-ITD+/DNMT3A+ (8.3 %). Patients with 1 mutation had a noticeably longer OS median compared with patients with 2 mutations (18.1 and 12.2 months; p = 0.003). In patients with NPM1+ according to their OS the most unfavorable additional mutation was FLT3-ITD (median 27.4 vs. 9.2 months; p = 0.019) and the combination of NPM1+/FLT3-ITD+/DNM-T3A+ (median 27.4 vs. 14.6 months; p = 0.141). OS of patients with DNMT3A+ showed a downward trend if FLT3-ITD additional mutation was identified (17.3 vs. 7.1 months; p = 0.074). Conclusion. Mutations in FLT3, DNMT3A, IDH1/2, NPM1 genes frequently occur in AML intermediate-risk patients, i.e. they determine the intermediate prognosis group in AML. The studied mutations considerably impact prognosis. It is important to take into consideration mutation type, its allele burden, and the presence of additional mutations. A patient with 2 mutations has a considerably worse OS compared with a patient with 1 mutation. The studied group of patients with the combination of NPM1+/FLT3-ITD+, NPM1+/ FLT3-ITD+/DNMT3A+, DNMT3A+/FLT3-ITD+ mutations has the poorest prognosis. Comprehensive analysis of genetic damages in AML patients allows to most accurately predict the course and prognosis of the disease and to plan targeted therapy.

AIMTo examine prognostic potential of the number of bone marrow (BM) blasts and cell karyotype as risk factors of transformation of myelodysplastic syndrome (MDS) in acute myeloblastic leukemia AML.MATERIAL AND METHODThe analysis of examination was made for 72 patients with primary MDS in the groups formed by number of blasts in BM, karyotype and IPSS variant. MDS was diagnosed by WHO criteria. Transformation into AML was established in blastosis > 20% in peripheral blood and/or BM. The karyotype was studied according to GTG technique.RESULTSMore frequent progression of MDS was seen in patients with blastosis > 10%, unfavourable karyotype and high IPSS risk. The least number of leukemic transformations occurred in karyotype of intermediate prognosis while disease-free survival in patients with karyotype of good prognosis was similar to that of patients with unfavourable karyotype. The number of blasts in BM and IPSS variant appeared to be prognostic markers of duration of leukemia-free survival in one-factor analysis. The multifactorial analysis found out one factor of MDS transformation in AML: number of blasts in BM puncture biopsy.CONCLUSIONPrognostic priority of the number of BM blasts as a risk factor of MDS progression compared to karyotype is explained by biological heterogenicity of MDS.

Aim. To characterize patients with mixed myeloid neoplasias with proliferation of neutrophils, platelets and eosinophils. Material and methods. Examination and treatment results were analysed for patients with atypical myeloid leukemia (n = 4), myelodysplastic syndrome (MDS, n = 1) and thrombocytosis, MDS and eosinophilia (n = I). The examination included morphological, histological, cytogenetic and molecular tests. Results. One patient with atypical chronic myeloid leukemia was prior diagnosed to have primarily MDS with a typical aberration of chromosome 5. Two other patients had an initial morphological picture of resistant anemia with blast excess, signs of myeloproliferation and extramedullar hemopoiesis. One and two months after the first examination they received transfusions of erythrocytic mass. Just then they were found to have splenomegaly and leukocytosis due to proliferating and maturating forms of neutrophils. The course of the disease in patients with MDS, thrombocytosis and normal karyotype and in patients with MDS, eosinophilia and combined chromosomal breaks including translocation (3;12)(q21;pl3) was characterized by resistance to standard programs of polychemotherapy and transformation into acute myeloblasts leukemia. Conclusion. In some cases atypical CML is a stage of a natural course of MDS. Some MDS variants with eosinophilia and thrombocytosis should be referred to the group of mixed myeloid neoplasias.