Varkani syndrome or trisomy 8 mosaicism (T8M) - is a described chromosomal anomaly with a frequency of 1 : 25,000 to 1 : 50,000 births, which is more common in men than in women (5 : 1). In the Department of Pediatric Surgery at Almazov NMRC, a 28-year-old woman was admitted at 39 weeks' gestation with a aggravated somatic history. Pregnancy occurred with the support of auxiliary reproductive technologies: extorporepical fertilization due to the male factor, overpermation and asthenospermia caused by mumps transferred in childhood. Antenatally diagnosed: multiple congenital malformations of the fetus: agenesis of the corpus callosum, triventricularhydrocephalus, macrocephaly, bilateral ureterohydronephrosis; megacystis. The spouses were karyotyped, karyotypes 46, XX - normal female and 46, XY - normal male. According to the results of the karyogram, the fetal karyotype is 46, XY - normal male. From the first day of life, the child has clinical epileptic myolonic generalized attacks. According to the results of the study: Neurosonography confirmed the presence of a malformation of the brain. Echocardiography: Ventricular septal defect (VSD). Ultrasound of the abdominal organs revealed bilateral ureterohydronephrosis with expansion of the pelvicalyceal system and ureters. At the age of 28 days, due to the existing neurogenic disorder of the function of the bladder against the background of a fixed spinal cord in order to constant urine, vesicostomy is performed. Performed diagnostic laparoscopy: in the right iliac region spreads education with a diameter of 2 cm, which is a diverticular-cystic doubling of the ileum. A wedge-shaped resection of the base of the diverticulum was performed, the operation was completed by vesicostomy. Discharged home in a satisfactory condition at the age of 2 months. As a result of karyotyping of the newborn, the result mos47, XY, +8[7]/46, XY [9] was obtained - a mosaic variant of trisomy on chromosome 8. Mosaicism of trisomy 8 rare chromosomal anomaly. The clinical case described by us is characterized by a combination of severe congenital defects, which were not previously reported. This is the first time of the birth of a child with a trisomy of 8 chromosome after auxiliary reproductive technologies: extracurporeal fertilization.

Aim. To assess the rate of DNMT3A, IDH1, IDH2, and ASXL1 gene mutations and their effect on the prognosis both as isolated findings and in combination with well-known chromosomal aberrations and gene mutations in newly diagnosed acute myeloid leukemia (AML) patients from some regions of the Russian Federation. Materials & Methods. The study enrolled 83 patients with newly diagnosed AML from 22 regions of the Russian Federation, who underwent molecular genetic examination for detecting IDH1 (R132), IDH2 (R140), ASXL1, and DNMT3A gene mutations with droplet digital PCR and Sanger sequencing methods. Results. The mutation rate in DNMT3A was 16.7 %, in IDH1 (R132) it was 6 %, in IDH2 (R140) it was 9.6 %, and in ASXL1 it was 6 %. The R140 mutation in IDH2 correlated with the older age of patients. The mutations in IDH1 (R132), IDH2 (R140), and DNMT3A showed a significant association with mutated NPM1. The mutations in IDH1 (R132), IDH2 (R140) were reported to occur significantly more often in patients with normal karyotype. The IDH1 (R132) and IDH2 (R140) mutations appeared to have a favorable effect on AML prognosis, which is most likely to be associated with a high rate of their compatibility with NPM1 mutation. The mutated type of DNMT3A had a negative effect on overall survival of patients with NPM1 mutation. The mutation in ASXL1 also appeared to be an unfavorable prognostic factor for overall survival of patients with wild type NPM1. Conclusion. A high rate of mutation occurrence in epigenetic regulation genes as well as the prognostic potential of these mutations in AML necessitate the need for determining the mutation status of DNMT3A, IDH1, IDH2, and ASXL1 in the context of primary diagnosis in real-world clinical practice.

Acute myeloid leukemias (AML) are the most ubiquitous of all adult leukemias. The prognosis of the disease depends on its genetic profile. The mutation in FLT3 gene, which codes FMS-like tyrosine kinase 3, is observed in 1/3 of patients and is responsible for a high rate of relapses. The prognosis of relapsed/refractory FLT3-positive AML is extremely poor. The standard intensive therapy rarely yields long-term responses. The new first- and second-generation FLT3 tyrosine kinase inhibitors enriched treatment opportunities for patients with this mutation. Gilteritinib, a potent second-generation FLT3-ITD/TKD inhibitor, is a new effective and well tolerated drug for the treatment of relapsed/refractory FLT3-positive AML. Due to its efficacy, low toxicity, and good manageability, this drug can be administered to all patients, including the elderly or those with severe comorbidities and complications of previous therapy. Besides, this drug can be used in outpatient units. The present paper contains three case reports dealing with different clinical situations in patients with FLT3-positive AML treated with gilteritinib in real-world clinical practice.

Internal tandem duplication is the most common form of mutation in FMS-like tyrosine kinase 3 (FLT3) in different haematological malignancies, highlighting in acute myeloid leukaemia (AML) and is associated with increased risk of relapse and reduced overall survival. A major breakthrough in the treatment of FLT3-mutated AML has been achieved through the use of highly selective FLT3 tyrosine kinase inhibitors, both in monotherapy and in combination with standard intensive cytotoxic chemotherapy. The desire to improve the outcomes of patients with AML, including those with relapse and refractory disease, has led to attempts to use non-standard therapeutic options. Enhancement of the antileukemic effects of the second-generation FLT3 inhibitor Gilteritinib may be achieved through synergy with the hypomethylating agent 5-azacytidine and the selective Bcl-2 inhibitor Venetoclax. Thus, targeted triple therapy is a promising option in the treatment of patients with FLT3-mutated AML. This study sought to evaluate the effectiveness of “triple therapy” regimen in 4 patients with relapsed/refractory FLT3 mutated AML. We found that the use of this combination showed rapid response with good safety and frequently allowed subsequent transplant and achieve durable clinical benefit.

Aim. To identify mutations in IDH1/IDH2, DNMT3A, and ASXL1 genes responsible for genome epigenetic regulation and their co-occurrence with FLT3, NPM1, and RUNX1 mutations in newly diagnosed adult acute myeloid leukemias (AML). Materials & Methods. The study included 56 patients with newly diagnosed AML treated at the VA Almazov National Medical Research Center. Among them there were 34 men and и 22 women aged 18-76 years (median 46 years). Mutation status of IDH1, IDH2, DNMT3A, and ASXL1 genes of epigenetic regulation was assessed by Sanger sequencing method. Molecular genetic analysis of FLT3, NPM1, and RUNX1-RUNX1T1 genes was performed using commercial kits. Results. Mutations in epigenetic regulation genes were detected in 14 (25 %) out of 56 patients. Mutation prevalence was not associated with risk groups (p = 0.072). IDH1/2 mutations were identified in 15.6 % of patients and were significantly oftener observed concurrent with NPM1 mutations (62.5 %; p = 0.01) compared to patients with wild-type IDH1/2. In most patients IDH1/2 mutations were associated with normal karyotype (p = 0.002). The DNMT3A (R882) mutation was identified in 4 (7.1 %) out of 56 patients within the analyzed group. In 6 patients (11.1 %) ASXL1 mutations were detected co-occurring with RUNX1-RUNX1T1 and FLT3-ITD mutations. Conclusion. Mutations in epigenetic regulation genes are often identified in AML patients and can be concurrent with abnormalities in NPM1, FLT3 и RUNX1 genes.

Цель. Выявление мутаций генов IDH1/IDH2, DNMT3A и ASXL1, ответственных за эпигенетическую регуляцию генома, при впервые диагностированных острых миелоидных лейкозах (ОМЛ) у взрослых и их сочетания с мутациями генов FLT3, NPM1, RUNX1. Материалы и методы. В исследование включено 56 пациентов с впервые выявленным ОМЛ, проходивших лечение в ФГБУ «НМИЦ им. В.А. Алмазова» Минздрава России. Среди них было 34 мужчины и 22 женщины в возрасте 18–76 лет (медиана 46 лет). Мутационный статус генов эпигенетической регуляции IDH1, IDH2, DNMT3A и ASXL1 определяли методом секвенирования по Сэнгеру. Молекулярно-генетический анализ генов FLT3, NPM1, RUNX1-RUNX1T1 выполняли с использованием коммерческих наборов. Результаты. Мутации генов эпигенетической регуляции обнаружены у 14 (25 %) из 56 пациентов. Распространенность мутаций не была связана с группами риска (p = 0,072). Мутации IDH1/2 выявлены у 15,6 % пациентов и статистически значимо чаще обнаруживались одновременно с мутациями NPM1 (62,5 %; p = 0,01) по сравнению с пациентами с диким типом IDH1/2. У большинства пациентов мутации IDH1/2 были связаны с нормальным кариотипом (p = 0,002). Мутация DNMT3A (R882) определена у 4 (7,1 %) из 56 пациентов анализируемой группы. У 6 (11,1 %) пациентов были идентифицированы мутации ASXL1, которые сочетались мутациями с RUNX1-RUNX1T1 и FLT3-ITD. Заключение. Мутации генов эпигенетической регуляции часто обнаруживаются у пациентов с ОМЛ и могут сочетаться с нарушениями в генах NPM1, FLT3 и RUNX1.

Миелоидная саркома (известная как хлорома или гранулоцитарная саркома) представляет собой редкое заболевание и характеризуется пролиферацией незрелых миелоидных клеток в экстрамедуллярных очагах поражения. Хлорома чаще развивается у пациентов с острыми миелоидными лейкозами, другими миелопролиферативными новообразованиями или миелодиспластическим синдромом, однако также может манифестировать в виде изолированной опухоли. Хотя миелоидная саркома может развиваться в различных органах и тканях, все же наиболее часто встречается поражение лимфатических узлов, мягких тканей и костей. Миелоидная саркома с первичным поражением женских половых органов описывается крайне редко. В литературе есть клинические наблюдения поражения шейки матки. В настоящей статье суммированы имеющиеся литературные данные, затрагивающие различные аспекты диагностики и лечения миелоидной саркомы. Обсуждается роль химиотерапии, лучевой терапии, хирургического вмешательства и трансплантации костного мозга в лечении данной злокачественной опухоли. Представляется, что оптимальным вариантом лечения миелоидной саркомы независимо от первичной локализации опухоли остается химиотерапия и трансплантация аллогенного костного мозга (аллоТКМ). Перспективным направлением в терапии является использование новых таргетных препаратов, способных улучшить результаты лечения. Мы представляем клиническое наблюдение пациентки с миелоидной саркомой шейки матки и сопутствующим вовлечением костного мозга, описываем особенности клинического течения, диагностики и лечения. Пациентка получила химиотерапию с последующей аллоТКМ. Проведенный объем предтрансплантационной терапии позволил выполнить аллоТКМ при максимально глубоком ответе. У пациентки достигнута ПЭТ- и МОБ-отрицательная полная ремиссия миелоидной саркомы шейки матки и костного мозга.

Aim. To assess the efficacy, safety, and tolerance of gemtuzumab ozogamicin (GO) combined with FLAG/FLAG-Ida chemotherapy or azacitidine in patients with relapsed/refractory acute myeloblastic leukemia (AML) in clinical practice. Materials & Methods. The study included 32 patients (16 men and 16 women). The median age was 44 years (range 23-83 years). Among them there were 15 (46.8 %) patients with refractory and 17 (53.2 %) patients with relapsed AML. GO combined with FLAG/FLAG-Ida was administered to 15 (46.8 %) patients, whereas 17 (53.2 %) patients were treated with GO and azacitidine combination. Therapy safety was assessed according to CTCAE v. 5.0. Results. Overall response rate including complete remission (CR), CR MRD-, CR with incomplete hematologic recovery, and morphologic leukemia-free status was 59.4 % (19/32). Refractoriness was observed in 31.25 % (10/32) of patients. Early mortality was 9.4 % (3/32). Overall response was 64.7 % (11/17) in the azacitidine and 53.3 % (8/15) in the FLAG/FLAG-Ida groups. In 4 (80 %) out of 5 patients with prior to FLAG treatment refractoriness, the response was achieved after GO + azacitidine therapy. In 58.9 % (10/17) of patients who received GO + azacitidine therapy, allogeneic hematopoietic stem cell transplantation (allo-HSCT) could be performed. The incidence of GO infusion complications in the tested groups did not significantly differ (p = 0.72) and was 46.7 % (7/15) (40 % with grade 1/2 and 6.7 % with grade 3) in the GO + FLAG/FLAG-Ida group and 35.3 % (6/17) (29.4 % with grade 1/2 and 5.9 % with grade 4) in the GO + azacitidine group. In the GO + FLAG/FLAG-Ida group 5 (33.3 %) patients experienced serious adverse events (SAE) of sepsis. In the GO + azacitidine group SAEs were reported in 6 (35.3 %) patients: 4 (66.6 %) with sepsis, 1 (16.7 %) with acute cardiovascular failure, and 1 (16.7 %) with acute respiratory failure. The median (range) duration was 23 (10-39) days for neutropenia grade 4, 24 (11-38) days for neutropenia grade 3, 21 (11-41) days for thrombocytopenia grade 4, 26 (16-45) days for thrombocytopenia grade 3, and 25 (22-45) days for thrombocytopenia grade 1/2. Thrombocytopenia duration was longer in patients with GO + FLAG/FLAG-Ida therapy, however, no significant differences were identified. No cases of veno-occlusive liver disease were reported. Median overall survival (OS) for both groups (n = 32) was 31.4 months, median disease-free survival (n = 21) was 13.3 months. In the group of patients with effective treatment, the median OS was not reached. In non-responders, it was 18 months (р = 0.0442). Conclusion. GO combined with FLAG/FLAG-Ida chemotherapy or azacitidine proved effective in relapsed/refractory AML patients. Remission did not appear to be associated with ELN risk, gender, age, CD33 expression, number of prior therapy lines, or number of relapses. GO + azacitidine combination showed efficacy, safety, and good tolerance in patients with prior high-dose chemotherapy refractoriness as well as low ECOG performance status. That allowed for the subsequent allo-HSCT administration to these patients. There was no significant difference between the groups of patients in the incidence of hematologic, non-hematologic toxicity, and time to hematologic recovery. Thrombocytopenia duration was longer in patients with GO + FLAG/FLAG-Ida therapy which is consistent with literature data. GO-based effective treatment in relapsed/refractory AML considerably improves OS: during 36 months of follow-up the median was not reached.

Цель. Оценить эффективность, безопасность и переносимость гемтузумаба озогамицина (ГО) в комбинации с химиотерапией FLAG/FLAG-Ida или азацитидином у пациентов с рецидивами и рефрактерным течением острого миелобластного лейкоза (ОМЛ) в реальной клинической практике. Материалы и методы. В исследование включено 32 пациента (16 мужчин, 16 женщин). Медиана возраста составила 44 года (диапазон 23–83 года). Среди них было 15 (46,8 %) пациентов с рефрактерным течением ОМЛ и 17 (53,2 %) — с рецидивами. ГО в комбинации с режимами FLAG/FLAG-Ida использовался у 15 (46,8 %) пациентов, в комбинации с азацитидином — у 17 (53,2 %). Безопасность терапии оценивалась согласно шкале CTCAE v. 5.0. Результаты. Частота общего ответа, включающего полную ремиссию (ПР), ПР МОБ–, ПР с неполным восстановлением показателей крови, морфологически свободный от лейкоза статус, составила 59,4 % (19/32). Рефрактерность наблюдалась у 31,25 % (10/32) пациентов. Ранняя летальность составила 9,4 % (3/32). Общий ответ в группе с азацитидином составил 64,7 % (11/17), с FLAG/FLAG-Ida — 53,3 % (8/15). У 4 (80 %) из 5 пациентов с рефрактерностью к предшествующему лечению по схеме FLAG достигнут ответ после терапии ГО + азацитидин. У 58,9 % (10/17) пациентов после терапии ГО + азацитидин удалось выполнить трансплантацию аллогенных гемопоэтических стволовых клеток (аллоТГСК). Частота инфузионных осложнений ГО в группах статистически значимо не различалась (p = 0,72) и составила 46,7 % (7/15) (40 % — I–II степени, 6,7 % — III степени) в группе ГО + FLAG/FLAG-Ida и 35,3 % (6/17) (29,4 % — I–II степени, 5,9 % — IV степени) в группе ГО + азацитидин. В группе ГО + FLAG/FLAG-Ida 5 (33,3 %) пациентов имели серьезные нежелательные явления (СНЯ) в виде сепсиса. В группе ГО + азацитидин СНЯ отмечались у 6 (35,3 %) пациентов: 4 (66,6 %) — сепсис, 1 (16,7 %) — острая сердечно-сосудистая недостаточность, 1 (16,7 %) — острая дыхательная недостаточность. Медиана (диапазон) длительности нейтропении IV степени тяжести составила 23 (10–39) дня, нейтропении III степени — 24 (11–38) дня, тромбоцитопении IV степени — 21 (11–41) день, III степени — 26 (16–45) дней, I–II степени — 25 (22–45) дней. Длительность тромбоцитопении была более продолжительной у пациентов, получавших ГО + FLAG/FLAG-Ida, однако статистически значимых различий не отмечено. Случаев веноокклюзионной болезни печени не зарегистрировано. Медиана общей выживаемости (ОВ) в целом по обеим группам (n = 32) составила 31,4 мес., безрецидивной (n = 21) — 13,3 мес. В группе пациентов с эффективным лечением медиана ОВ не достигнута. В группе без эффекта этот показатель составил 18 мес. (p = 0,0442). Заключение. Комбинации ГО с химиотерапией FLAG/FLAG-Ida или азацитидином оказались эффективными у пациентов с рецидивами и рефрактерным течением ОМЛ. Возможность достижения ремиссии не зависела от принадлежности к группе риска по ELN, пола, возраста, уровня экспрессии CD33, числа предшествующих линий терапии, количества рецидивов. Комбинация ГО + азацитидин оказалась эффективной, безопасной и вполне переносимой у пациентов, рефрактерных к предшествующей высокодозной химиотерапии, а также при отягощенном соматическом статусе по ECOG. Это позволило в последующем выполнить аллоТГСК у данной категории больных. Частота развития гематологической и негематологической токсичности, длительность восстановления показателей крови статистически значимо не различались в обеих группах. Тромбоцитопения была более продолжительной в группе ГО + FLAG/FLAG-Ida, что согласуется с литературными данными. Эффективное лечение на основе ГО при рецидивах и рефрактерном течении ОМЛ существенно улучшает показатели ОВ: при сроке наблюдения 36 мес. медиана не достигнута.

Myeloid sarcoma, also known as chloroma or granulocytic sarcoma, is a rare disease characterized by the proliferation of immature myeloid cells in extramedullary lesions. Chloroma is more commonly observed in patients with acute myeloid leukemias, other myeloproliferative neoplasms, or myelodysplastic syndrome. However, it can also manifest itself as solitary tumor. Sarcoma can develop in different organs and tissues, but most frequently it appears in lymph nodes, soft tissues, and bones. Myeloid sarcoma with primary gynecological lesion is very rarely mentioned. In literature cases of cervical lesions are described. The present article summarizes the literature data concerning different aspects of myeloid sarcoma diagnosis and treatment. The issue under discussion is the role of chemotherapy, radiotherapy, surgery, and bone marrow transplantation in the treatment of this malignant tumor. It appears that whatever the primary tumor localization, the best treatment options are chemotherapy and allogeneic bone marrow transplantation (allo-BMT). A promising trend is the use of novel targeted drugs improving outcomes of treatment. The article provides a case report of a female patient with cervical myeloid sarcoma and concomitant bone marrow involvement, as well as the description of clinical course, diagnosis, and treatment. The patient received chemotherapy with subsequent allo-BMT. The pre-transplant therapy enabled allo-BMT with the deepest response possible. The patient achieved PET-and MRD-negative complete remission of cervical myeloid sarcoma and bone marrow.

Aim. To assess the efficacy of FLAG/FLAG-Ida regimen and to identify factors that influence remission, duration of disease-free survival (DFS) and overall survival (OS) of patients with relapsed/refractory acute myeloid leukemia (AML). Materials & Methods. The trial included 54 patients (28 men and 26 women), median age was 37 years (range 18–70 years). 27 (50 %) out of 54 patients had refractory AML and 27 (50 %) patients had relapsed AML. FLAG and FLAG-Ida regimens were administered as induction therapy. 37 (68.5 %) patients received bone marrow transplantation. Molecular genetic and cytogenetic examinations were performed prior to therapy and on the 28th day after the start of treatment. WT1 gene expression was evaluated on the 14th–16th day of treatment. Results. Complete remission (CR) was achieved in 42 (77.8 %) out of 54 patients. Refractoriness to therapy was observed in 9 (16.7 %) out of 54 patients, mortality was 5.5 % (3/54). Remission rate was higher in patients with relapsed AML compared with refractory AML: 85.2 % (23/27) and 70.4 % (19/27), respectively. On the 14th–16th day of treatment patients with blast cell count ≥ 10 % in bone marrow (BM) showed significantly lower CR rate (60 %) compared with the group of patients with < 10 % blast cells in BM (89.6 %; p = 0.024) and shorter DFS (median 7.6 vs. 17.6 months, respectively; p = 0.03). Median DFS in patients with WT1 expression reduction to < 1 log on the 14th–16th day was 5 vs. 18 months in patients without WT1 expression reduction (p = 0.01). DFS varied in groups of patients with blast cell count < 10 % in BM on the 14th–16th day of treatment based on the level of WT1 expression reduction (p = 0.04). MRD-negative patients (57.1 %) showed significantly longer DFS and OS compared with MRD-positive patients (median DFS was 17.6 vs. 5.2 months, respectively, p = 0.02; median OS was 19 vs. 6.9 months, p = 0.0002). Median DFS and OS were different only in ELN low- and high-risk groups (median not reached vs. 5.2 months, respectively, p = 0.039; median not reached vs. 10.2 months, p = 0.039). Conclusion. FLAG and FLAG-Ida are effective and safe regimens in the treatment of relapsed/refractory AML. Achieving remission depends on neither the risk group nor the time of relapse occurrence. The blast cell count in BM on the 14th–16th day of FLAG/FLAG-Ida treatment is a prognostic factor determining achievement and duration of remission. WT1 expression level in the early post-induction period is a sensitive DFS marker. MRD status and molecular genetic risk (ELN) group affiliation are essential prognostic factors determining DFS and OS.

Aim. To assess the efficacy of FLAG/FLAG-Ida regimen and to identify factors that influence remission, duration of disease-free survival (DFS) and overall survival (OS) of patients with relapsed/refractory acute myeloid leukemia (AML). Materials & Methods. The trial included 54 patients (28 men and 26 women), median age was 37 years (range 18-70 years). 27 (50 %) out of 54 patients had refractory AML and 27 (50 %) patients had relapsed AML. FLAG and FLAG-Ida regimens were administered as induction therapy. 37 (68.5 %) patients received bone marrow transplantation. Molecular genetic and cytogenetic examinations were performed prior to therapy and on the 28th day after the start of treatment. WT1 gene expression was evaluated on the 14th-16th day of treatment. Results. Complete remission (CR) was achieved in 42 (77.8 %) out of 54 patients. Refractoriness to therapy was observed in 9 (16.7 %) out of 54 patients, mortality was 5.5 % (3/54). Remission rate was higher in patients with relapsed AML compared with refractory AML: 85.2 % (23/27) and 70.4 % (19/27), respectively. On the 14th-16th day of treatment patients with blast cell count ≥ 10 % in bone marrow (BM) showed significantly lower CR rate (60 %) compared with the group of patients with < 10 % blast cells in BM (89.6 %; p = 0.024) and shorter DFS (median 7.6 vs. 17.6 months, respectively; p = 0.03). Median DFS in patients with WT1 expression reduction to < 1 log on the 14th-16th day was 5 vs. 18 months in patients without WT1 expression reduction (p = 0.01). DFS varied in groups of patients with blast cell count < 10 % in BM on the 14th-16th day of treatment based on the level of WT1 expression reduction (p = 0.04). MRD-negative patients (57.1 %) showed significantly longer DFS and OS compared with MRD-positive patients (median DFS was 17.6 vs. 5.2 months, respectively, p = 0.02; median OS was 19 vs. 6.9 months, p = 0.0002). Median DFS and OS were different only in ELN low- and high-risk groups (median not reached vs. 5.2 months, respectively, p = 0.039; median not reached vs. 10.2 months, p = 0.039). Conclusion. FLAG and FLAG-Ida are effective and safe regimens in the treatment of relapsed/refractory AML. Achieving remission depends on neither the risk group nor the time of relapse occurrence. The blast cell count in BM on the 14th-16th day of FLAG/FLAG-Ida treatment is a prognostic factor determining achievement and duration of remission. WT1 expression level in the early post-induction period is a sensitive DFS marker. MRD status and molecular genetic risk (ELN) group affiliation are essential prognostic factors determining DFS and OS.

Background. In patients with chronic lymphocytic leukemia (CLL) the eradication of minimal residual disease (MRD) is a prognostic factor of overall survival (OS) and progressionfree survival (PFS). IGHV mutational status has also independent prognostic value. Aim. To analyse the impact of mutational status and MRD eradication in CLL patients after first-line standard BR (ben-damustine + rituximab) immunochemotherapy. Materials & Methods. The prospective study included patients with immunophenotypically confirmed CLL who had not previously received anticancer therapy. All patients were treated by BR combination from 2012 to 2015. MRD level was determined in 109 patients after completing the 3rd and the 6th treatment courses. IGHV mutational status data were available for 98 patients. IGHV mutational status was evaluated in accordance with ERIC recommendations. MRD was assessed by standardized method of 4-color flow cytometry. Results. MRD negativity was achieved in 37 (34 %) out of 109 patients. MRD eradication correlated with the best PFS (p = 0.04). IGHV mutational status had a statistically significant impact on PFS (p = 0.02). In patients with MRD-nega-tive response and IGHV mutation no unfavorable events occurred during the period of monitoring. Conversely, PFS rates in MRD-negative patients having no IGHV mutation and in MRD-positive patients with mutation were significantly worse. MRD eradication resulted in statistically significant improvement of PFS rates after completing 3 treatment courses, compared with the cases with MRD persistence regardless of residual malignant clone level (p = 0.01). Conclusion. BR therapy as first-line treatment statistically improved PFS in patients who achieved MRD-negative remission after completing the 3rd treatment course. PFS was significantly higher in MRD-negative patients with IGHV mutation after 6 treatment courses. MRD negativity resulting from 6 BR therapies in patients having no / GHV mutation was not accompanied by PFS improvement. It follows that by itself MRD negativity cannot be considered to be a universal prognostic factor.

Background. Acute myeloblastic leukemia (AML) with NPM7 mutation amounts to 30 % of all AML and is characterized by good prognosis with the exception of cases with FLT3-/TD mutation. Despite the good prognosis, the likelihood of relapses in patients with NPM7 mutation may significantly differ. Thus, the estimation of the minimal residual disease (MRD) after chemotherapy and during follow-up is becoming increasingly important. This approach will make it possible to predict the sensitivity of a tumoral clone to chemotherapy. Aim. To evaluate the prognostic value of highly specific marker (NPM7 mutation) and non-specific marker (WT1 overexpression) of MRD, as well as to identify the correlation between the levels of NPM7 and WT7 at different stages of therapy and in the follow-up period. Materials & Methods. The research included 14 patients with AML. All patients had the NPM7 mutation and WT7 overexpression: 50 % of patients had additional molecular markers (BAALC overexpression, FLT3-/TD, DNMT3A, and MLL mutations). Real-time PCR was used for long-term monitoring of WT7 expression levels and NPM7 mutation. Results. The median decrease of NPM7 levels after the induction therapy was 3 log. All patients had relapses, NPM7 mutation, and lower rates of OS/RFS, which significantly correlated with prognostically negative molecular markers. There were no statistically significant differences in RFS in groups with the decrease of WT7 expression level < 2 log and ≥ 2 log on day 28 of treatment. At the same time, the decrease of WT7 expression by > 2 log was associated with significant differences in early relapses, which correlated with the decrease of NPM7 levels (> and < than 3 log) is revealed. RFS rates were higher in patients with WT7 expression level of < 100 per 104 copies ABL on day 28 and WT7 of < 250 per 104 copies ABL on day 14 of treatment. WT7 expression was significantly lower on days 14 and 28 in patients with NPM7 decrease of > 3 log on day 28. The decrease in WT7 expression of < 100 per 104 copies ABL on day 28 was more common in patients with isolated NPM1 mutation, compared to patients with additional negative molecular markers. Conclusion. The decrease in NPM1 levels after the induction therapy may serve as reliable prognostic marker of RFS and OS rates. New correlation between the degree of NPM1 reduction and the presence of additional molecular markers was established. Highly specific (NPM1 mutation) was shown to be more specific compared to non-specific markers ( WT1 overexpression). The research showed the predictive value of a lower limit level of WT1 on day 28 of treatment (100 per 104 copies ABL), and for the first time, the importance of the early assessment WT1 expression reduction on day 14 of induction therapy.

Background & Aims. Flow cytometry is successfully used for diagnosis of malignant lymphoproliferative disorders. However, there are atypical cases that are difficult to interpret; thus, new markers relevant for the differential diagnosis are to be searched for. The aim is to analyze CD200 expression in patients with B-cell lymphoproliferative disorders. Materials & Methods. 187 patients with chronic lymphocytic leukemia (CLL), 14 patients with mantle cell lymphoma (MCL), 9 patients with marginal zone lymphoma (MZL), and 5 patients with hairy cell leukemia (HCL) were enrolled in the study. Neoplasm was not confirmed in 12 subjects. The patients underwent the following tests: CBC, immunophenotyping of peripheral blood or bone marrow lymphocytes, and a cytogenetic test. In some cases, an additional immunohistochemical test of bone marrow trepanobiopsy or lymph node biopsy samples was required. Results. In all cases of CLL and HCL, the CD200 expression was positive; mean fluorescence intensity was higher in these cases as compared to other groups. Negative expression of CD200 prevailed in MCL patients; however, at the same time 2 cases of intermediate and positive expression were reported, both showing moderate fluorescence intensity values. CD200 expression was heterogeneous in MZL patients. Conclusion. The CD200 negative expression excludes typical HCL and CLL. Additional cytogenetic and immunnohistoсhemical tests should be performed in such cases to verify the diagnosis, first of all, MCL or MZL.

In present research the comparative analysis of donor chimerism (DC) using different tests was performed to improve the diagnostic tool in patients with malignant hematological disorders after allo-HSCT. The RBC antigen typing, identification of ABO blood type and quantitative analysis of InDel-, STR-, Y-polymorphisms were carried out for detection of DC. In addition, the expression of well-known oncogenes and CD-markers for monitoring MRD was evaluated to predict relapse and clinical outcome. According to our research, the analysis of InDel polymorphism using AlleleSEQR-PCR is more sensitive test for estimation of DC as compared with other assays. Moreover, the sensitivity of AlleleSEQR-PCR may be increased after isolation of the CD34 cell population in bone marrow. Nevertheless, observation of high levels in DC (³95%) in some leukemia patients (ALL, Ph+, bcr-abl/p190+) during first 6 months after HSCT cannot exclude the possibility of relapse. Thus, the combined monitoring of both DC (InDel) and MRD (oncogenes, WT1 and CD-markers) is a more advisable and useful test in managing hematologic malignancies and predicting relapse risk after allo-HSCT.

At present, cytogenetic and PCR-based techniques are generally used in the diagnosis of leukemia. Due to high accuracy, specificity, and sensitivity of PCR assays, they have the advantage over traditional cytogenetic tools. As a rule, the classical real time PCR is carried out in a 25-μL reaction mixture. It requires a large volume of each reagent and takes a long time to finish the test. The molecular genetic assay presented here is microchip-based real-time PCR that is optimized for simultaneous analysis of 15 oncogene mutations and one housekeeping gene abl . Moreover, this diagnostic tool requires a minimal amount of cDNA and PCR reagents (10-fold less) and a short time (2-fold less) for quantitative estimation of more than five copies of gene-target. Thus, microchip-based PCR analysis can improve the detection of oncogene mutations in leukemia patients and may be used for both early diagnostics and long-term monitoring of leukemia.