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The main goal is to optimize hematopoietic stem cell transplantation (HSCT) approach among adolescents and young adults with paroxysmal nocturnal hemoglobinuria (PNH) by means of Graft-versus-host disease (GVHD) and post-transplant complications risk lowering. Materials and methods. We report our experience of HSCT from HLA-matched unrelated donors using TCR alfa/beta and CD19 depletion in 5 pts (1M/4F) with PNH, developed after successful immunosuppressive therapy (IST) of acquired aplastic anemia (AA). Median age of pts at the moment of transplantation was 17,8 years (range 14,5-22,7), median interval from IST to PNH was 4 years (5mo - 6,5 y). In all patients non-severe pancytopenia was present: granulocytes 0,8х109/l (0,8-1,8 х109/l) platelets 106 х109/l (27-143 х109/l) and Hb -78 g/l, median PNH clone size in granulocytes was 94 (range 75-99)%. One pts previously developed sinus thrombosis. Conditioning consisted of thoraco-abdominal irradiation 4-6 Gy, cyclophosphamide 100 mg/kg, fludarabine 150 mg/m2 and anti-thymocyte globulin (ATG) or alemtuzumab. Eculizumab was given from day (-7) till day (+14) (every 7 days, only 4 times). GVHD prophylaxis was tacrolimus ± methotrexate. Results. Infusedgraft characteristics were: CD34+ - 8,1х106/kg, CD3TCRab·150х103/kg, CD3gd+ - 7,3х106/kg, СD19+ - 221х103/kg, NK -6,4х108/kg. Engraftment was achieved in all 5 pts with a median of 15(12-18) и 13(10-18) days for granulocytes and platelets, respectively. Skin acute GVHD grade I developed in only 1 pt, and subsided with short course of glucocorticoids. CMV reactivation occurred in 1 pt; there were no episodes of Epstein-Barr Virus (EBV) o rAdenovirus (AdV) reactivation. Full donor myeloid chimerism was established in all pts by day +30. Immune reconstitution was delayed until 6 months after transplant but no severe infections occurred. All pts are alive 1,7-5,5 years (med 4 years) after HSCT with normal hematopoiesis and immune function, full donor chimerism and no late sequelae. Conclusions. Transplantation of TCRalfa/beta and CD19 depleted hematopoietic cells from matched unrelated donor after immunoablative conditioning and supported with short course of eculizumab is perfectly safe and efficient technology leading to cure in young patients with PNH.

Aim. To evaluate the safety and efficacy of crizotinib used in pediatric patients with relapsed or refractory ALK-positive anaplastic large-cell lymphoma (ALCL). Subjects and methods. The paper describes the experience with crizotinib used in 8 patients with refractory ALK-ALCL before and after allogeneic hematopoietic stem cell transplantation (HSCT). Results. All the 8 (100%) patients treated with crizotinib were recorded to have complete responses, including complete metabolic ones (tumor disappearance as evidenced by positron emission tomography (PET)/computed tomography. Conclusion. Low and manageable toxicity of crizotinib and complete PET-negative responses in patients with resistant ALK lymphomas favor the need to test the drug as first-line therapy, by possibly decreasing the intensification of chemotherapy.

Aim. To determine predictors for decision-making on a differential approach to choosing glucocorticosteroids (GCS) for children and adolescents with acute lymphoblastic leukemia (ALL). Subjects and methods. The analysis covered 1064 primary patients aged to 1 to 18 years with ALL who had been registered at the clinics of Russia and Belorussia in April 2002 to November 2006. Before induction therapy, the patients were randomized into a dexamethasone (DEXA) 6 mg/m2 group (n=539) and a methylprednisolone (MePRED) 60 mg/m2 one (n=525). Results. The entire group showed no statistically significant differences in survival rates between the patients receiving DEXA or MePRED. However, an analysis of age groups revealed the benefits of DEXA in children younger than 14 years (the event-free survival (EFS) was 76±2 and 71±2%, respectively (p=0.048); the overall survival (OS) was 81±2 and 77±2%, respectively (р=0.046); therapy-induced mortality was 6.4% (DEXA) and11.1% (MePRED) (р=0.014); the rate of isolated extramedullary relapses was 1.5% (DEXA) and 4.4% (MePRED) ( р=0.009). At the same time, EFS and OS in 14-to-18-year-old adolescents were statistically significantly higher than in those who used MePRED (EFS, 65±6 and 52±6%, respectively (р=0.087); OS, 72±6 and 61±6%, respectively; (р=0.17). Conclusion. The findings suggest that it is possible that the choice of a GCS for ALL therapy must be also based on a patient’s age. There is a need for further studies of this matter in prospective randomized multicenter trials in children and adolescents.

Aim. To analyze the specific features of recurrences of acute promyelocytic leukemia (APL) in children after standard therapy with daunorubicin, cytosine arabinoside (Ara-C), all-trans retinoic acid (ATRA) and to develop further programmed treatment policy. Subjects and methods. The study included 9 patients with recurrent APL. The recurrences developed significantly more frequently in a very high-risk group (patients with minimal residual disease being preserved after the intensive therapy phase). Induction used arsenic trioxide (ATO) and/or standard chemotherapy + ATRA; ATO monotherapy was in consolidation. CD34+ cells were mobilized until molecular remission was achieved with high-dose Ara-C and granulocyte colony-stimulating factor. Pretransplantation conditioning involved melfalan as a basic drug in combination with high-dose Ara-C (5 pts), treosulfan (1 pt) or bisulfan (1 pt). Six patients received gemtusumab ozogamicin, 3-9 mg/m2, at different stages of therapy. Results. Before therapy one patient died; 8 patients achieved the second molecular remission; CD34+ cell mobilization and sampling were effective in 7 cases. Five patients were in long-term molecular remission after autologous hemopoietic stem cell transplantation (autoHSCT). Follow-up was 23-40 months. One patient is being prepared for transplantation. Following autoHSCT, another patient with a developed repeat recurrence died from complications due to related partially compatible transplantation. Visceral, including cardiological, toxicity of therapy was insignificant. In the APL-2003 protocol, overall and event-free survival rates were 93±3 and 76±6%, respectively. Conclusion. The application of ATO and autoHSCT in recurrent APL makes it possible to achieve and preserve the second molecular remission in case of insignificant extrahematological toxicity. Russian clinics should have access to ATO.

Aim. To analyze the results of allogeneic and autologous hemopoietic cell transplantations (allo- and auto-HCT) in children with acute myeloid leukemia (AML) from an intermediate risk group, most of which were performed using lower-intensity conditioning modes. Subjects and methods. The study enrolled 36 children from an intermediate risk group, who had undergone auto-HCT (n = 22) or allo-HCT (n = 14) in December 1994 to December 2008. The patients' age was 0.7 to 16.6 years (median 12.8 years). Chemotherapeutic conditioning regimens were applied to all the patients. Melphalan was a basic myeloablative agent in 83.3% of cases. Results. With a median follow-up of 4.6 years (1.1-13.8 years), three-year relapse-free survival (RFS) was 80.4%; overall survival (OS) was 65.6%. Recurrences were documented only in 6 (16.6%) patients from the auto-HCT. Transplantation-associated mortality (TAM) was 13.8% (five patients died). After allo-HCT versus auto-HCT, RFS, OS, and TAM were 100 and 68.7% (p = 0.03), 93.2 and 55.5% (p = 0.02), and 7.1 and 18.2%, respectively. Acute and chronic graft-versus-host reactions developed in 57.1 and 23.1%, respectively. Conclusion. Transplantation of allogeneic hemopoietic cells from a compatible related donor in the intermediate risk group children with AML, by using melphalan-based conditioning regimen, demonstrates a high survival rate with the minimum toxicity.

Aim. To analyze the efficiency of transplantation of the bone marrow from a HLA-compatible unrelated donor and continued immunosuppressive therapy (IST) in children with aplastic anemia (AA) unresponsive to 2 courses of IST. Subjects and methods. The study enrolled 14 children aged 2-16 years (median 9 years). A control group comprised 26 patients in whom IST was continued. The median interval between the diagnosis of AA and transplantation was 26 months (9-156 months). The conditioning regimen consisted of thoracoabdominal irradiation in a dose of 2 Gy, fludarabin (Flu) 100-150 mg/m2, cyclophosphamide (Cy) 100-200 mg/kg, antithymocyte globulin (ATG) in 11 patients and Flu, Cy, and ATG in 3. A graft-versus-host reaction was prevented with mycophenolate mefetil in all the patients, tacrolimus in 11, and cyclosporin A in 3. Donors were compatible for high-resolution typing of 10/10 and 9/10 alleles in 8 and 6 patients, respectively; the source of a transplant was bone marrow in 13 patients and granulocyte colony-stimulating factor-mobilized peripheral blood precursors in one case. Results. Thirteen patients achieved primary engraftment after single transplantation; one patient did after repeat transplantation. Grades I to II graft-versus-host reaction (GVHR) developed in 9 patients; postengraftment life-threatening infections in 3, extensive chronic GVHR in 2, circumscribed GVHR in 7. All fourteen hemopoietic cell transplant recipients followed for a median 17.5 months (range 1-71 months) were survivors. Conclusion. The likelihood of good survival after unrelated transplantations in AA is much higher than that after continued IST: 100% versus 15±11%.

Abstract Juvenile myelomonocytic leukemia (JMML) is a rare myeloproliferative disease of early childhood that develops due to mutations in the genes of the RAS-signaling pathway. Next-generation high throughput sequencing (NGS) enables identification of various secondary molecular genetic events that can facilitate JMML progression and transformation into secondary acute myeloid leukemia (sAML). The methods of single-cell DNA sequencing (scDNA-seq) enable overcoming limitations of bulk NGS and exploring genetic heterogeneity at the level of individual cells, which can help in a better understanding of the mechanisms leading to JMML progression and provide an opportunity to evaluate the response of leukemia to therapy. In the present work, we applied a two-step droplet microfluidics approach to detect DNA alterations among thousands of single cells and to analyze clonal dynamics in two JMML patients with sAML transformation before and after hematopoietic stem cell transplantation (HSCT). At the time of diagnosis both of our patients harbored only “canonical” mutations in the RAS signaling pathway genes detected by targeted DNA sequencing. Analysis of samples from the time of transformation JMML to sAML revealed additional genetic events that are potential drivers for disease progression in both patients. ScDNA-seq was able to measure of chimerism level and detect a residual tumor clone in the second patient after HSCT (sensitivity of less than 0.1% tumor cells). The data obtained demonstrate the value of scDNA-seq to assess the clonal evolution of JMML to sAML, response to therapy and engraftment monitoring.

In children, therapy-related hematologic neoplasms (t-HN) are uncommon. Many are driven by genetic events independent of clonal hematopoiesis. We sought to understand the clinical and genetic factors of pediatric t-HN in a large independent cohort. Fifty-six t-HN were retrospectively identified. Chromosome microarray, next-generation and/or RNA sequencing were performed. Patients had primary hematologic, solid, or central nervous system tumors. t-HN included myeloid (t-MN) and lymphoblastic (t-ALL) phenotypes. Approximately half of the cases harbored KMTA2A rearrangement (KMT2Ar). Among t-HN without KMT2Ar, genetic drivers were heterogeneous, including diverse fusions or aneuploidy. Approximately 18% harbored 17p deletions and/or TP53 mutations. EFS/OS was not associated with t-HN lineage or KMT2Ar, but HSCT was associated with improved EFS and OS. We detail one of the largest cohorts to date of pediatric t-HN, confirming frequent KMT2Ar and t-ALL. [ABSTRACT FROM AUTHOR]

Background: To date, fewer than 20 patients have been identified as having germline biallelic mutations in the coronin-1A gene (CORO1A) and its protein with clinical features of combined immunodeficiency characterized by T-cell lymphopenia ranging from the severe phenotype to the mild phenotype, recurrent infections, and lymphoproliferative disorders. However, the effects of CORO1A protein disruption on actin-dependent functions in primary cells have not been fully delineated. Objective: We sought to characterize the underlying defects of actin-dependent cellular functions in a female patient with combined immunodeficiency caused by a novel missense variant in the CORO1A gene in combination with a de novo heterozygous microdeletion of chromosome 16p11.2 and also to provide evidence of the pathogenicity of this gene mutation. Methods: To identify the genetic defect, next-generation sequencing followed by Sanger confirmation and array comparative genomic hybridization were performed. Western blot and quantitative PCR tests were used to assess the effects on the protein. Flow cytometry and live microscopy were performed to investigate cellular motility and immune cell counts and function. Results: We demonstrated that the CORO1A hemizygous variant c.19C>T, p. A7C induces significant decreases in cellular levels of the CORO1A protein while leaving mRNA concentrations unaffected. The observed mutation resulted in impaired natural killer cell cytotoxicity and platelet calcium signaling. In addition, primary granulocytes and mesenchymal cells showed significant defects in motility. Conclusion: Collectively, we added new data about the CORO1A gene as a key player in actin cytoskeleton dynamics and cell signaling. Our findings expand the clinical spectrum regarding CORO1A protein deficiency and confirm the importance of a personalized therapeutic approach for each patient.

Protocol AML-MM-2000 treatment results of unfavorable prognostic group in children with primary acute myeloid leukemia (AML) in Russia and Belarus are presented. 105 children at the age from 2 weeks till 18 years (a median age — 10.8 years) are included in the study. In 91 patients (86.7%) hematological remission have reached, from them 34 patients (37.4%) are in the first continuous complete remission (CCR). 6-years overall survival (OS), event-free survival (EFS) and relapse-free survival (RFS) rate for all group of patients was 0.35 ±} 0.05; 0.32 ±} 0.04 and 0.43 ±} 0.05, respectively. Survival analysis of children with different cytogenetic anomalies has allowed to define two prognostic groups of patients: the intermediate prognosis, including patients with normal kariotype and t(9;11) which EFS and RFS rate was 40—50%, and unfavorable prognosis with EFS and RFS rate less than 40%. The efficiency analysis of three postremission therapy type (allogeneic and autological hematopoietic stem cells transplantation (HSCT) and chemotherapy (HT)) in intermediate and unfavorable prognostis groups was carried out. High efficiency of related HLA-identical HSCT in patients both intermediate and unfavorable prognostis groups have been shown. Autological HSCT is possible in intermediate prognosis patients with absence of HLA-identical siblings and in unfavorable prognosis patients with absence of HLA-identical unrelated donor.

Prognosis of patients with juvenile myelomonocytic leukemia not undergoing allogeneic hematopoietic cells transplantation (SCT) is dismal and not affected by any cytotoxic chemotherapy. Differentiating therapy is an attractive approach in JMML, due to ability of JMML cells to differentiate and die under influence of treatment with various agents in vitro. We report results of first-line treatment of 14 patients with JMML with 13-cis retinoic acid (RA) 100 mg/m2 daily peroral intake and low dose Ara-C - 25 mg/m2 daily for 10 days monthly. Complete remission (CR) or hematological improvement (HI) was obtained in 7 patients (50%). Four patients (median age at disease manifestation 5 months, median level of HbF —6,2%) achieved CR after a median of 9 cycles of therapy; in 3 patients the best response was HI (age at disease manifestation - 5; 7 and 46 months, HbF — 4,2; 8,0 and 51,7%) after 2; 3 and 5 cycles. Three out of 4 patients with CR are alive and have been in CR for 10; 17 and 30 months after start of therapy, acute lymphoblastic leukemia developed in one patient after 84 months of CR. Two patients with HI are still receiving treatment and have been in stable condition for 10 months after the initiating therapy, while 1 patient died after 8 months of therapy from catheter-related sepsis. The progression of the disease despite the treatment was registered in 7 patients (median age at disease manifestation 16,3 months, median HbF — 35,4%). We conclude that 13-cis RA in combination with low dose AraC may be useful in the treatment of patients younger than 1 year with low level of HbF, although the general paradigm of treatment of JMML with SCT remains unchanged.

The use of all-trans retinoic acid (ATRA) drastically improved the results of therapy for acute promyelocytic leukeimia (APL). The high efficiency of the Russian protocol APL-93—98 applied to 62 children and adolescents with APL (event-free and overall survival (EFS and OS) were 84±5% respectively with a relapse rate of 7%) was achieved, by administering the high cumulative dose of daunoruicin (495 mg/m2) and single doses of АТRА (45 mg/m2). In this connection, the APL-2003 protocol attempted to reduce both acute and chronic toxicity of the therapy, by decreasing the cumulative dose of daunorubicin to 405 mg/m2 and the single dose of АТRА to 25 mg/m2 under regular control of minimal residual disease via molecular monitoring of the specific transcript PML/RARα. Analysis of the results of treatment in 61 patients aged 1.3 to 17 years (median 11.3) showed that despite of the reduced protocol intensity, the efficiency of therapy did not generally reduce — EFS and OS were 79±6 and 93±3%, respectively. The likelihood of the development of a relapse was significantly affected by molecular resistance, i.e. the retention of the transcript PML/RARα before a phase of maintenance therapy: the recurrence risk in this group was 57% whereas there were no differences in all cure rates between the patients depending on baseline leukocytosis. All 6 patients with relapses were treated with arsenic trioxide, resulting in not only clinical and hematological, but also molecular remissions; thereafter the patients received myeloablative therapy and autologous hemopoietic stem cell transplantation. The second remission lasted 3 to 24 months. Thus, reducing the cumulative dose of anthracyclines and the dose of АТRА did not make the results of the therapy for APL worse. The monitoring of minimal residual disease before and during maintenance therapy is a mandatory component of successful treatment in APL patients. Arsenic trioxide is an effective agent for the treatment of APL relapses.

Standard therapy of high risk patients with Langerhans cell histiocytosis (LCH) is associated with essential risk of therapy failure. In the present study results of 2-clorodeoxyadenosine usage as alternative therapy in this group of patients are analyzed. In this study 31 patients with multisystem form LCH with involving of organs at risk were included. Nineteen patients have received Prednisone and Vinblastine as front line therapy (standard group). Nine patients have received therapy with 2-CdA and cytosine-arabinoside (group 2-CdA). The patients in standard group statistically significant more often (55.5 % vs. 0 %, р = 0.0095) required second line therapy. Permanent disease conse-quences was more often developed in standard group (63.6 % vs. 0 %, р = 0.0147). The overall survival was 65.7 ± 12,8 % among standard group patients and 88.8% ±10% in 2-CdA group; p = 0.27. The event-free survival was in standard group 33.9 ± 12 %, in 2-CdA group – 88.8 ± 10 %, p = 0.0207. Severe hematological toxicity was observed at 2-CdA therapy. 2-CdA therapy is highly effective in treatment of high risk patients with LCH. The drug should be used only in a specialized hospital.

Introduction of biosimilars in clinical practice results in substantial reduction of direct medical costs. However, limited data concerning safety and tolerance of biosimilars exist, especially in children. We conducted double blind randomized trial comparing immediate tolerance of locally manufactured filgrastim (LeucostimR, “Biocad”, Russia) with original formulation (“Neupogen”, “Roche”, Switzerland) in children with various hematologic and oncologic diseases. Ten patients (4 F; 6 M) with a median age of 9.5 y (range 5—16 y) in whom G-CSF was indicated either by protocol or clinically were included into the study. All patients were scheduled to receive both formulations of G-CSF in order determined by randomization with permuted blocks. Pain was assessed with visual analog scale, ranging sensation of pain from 0 to 10 points. There were 41 injections of LeucostimR and 39 of Neupogen, and а median number of points attributed was 1.134 (0—10), and 1.661 (0—9) respectively (p=0,669). Intrapatient pain scores were quite consistent and did not differ by more than 1 point between any two injections. Other immediate toxicities were negligible or inexistent. Our study clearly demonstrates that both formulation of filgrastim are equally well tolerated.

Donor lymphocyte infusion has been used in the management of relapsed hematologic malignancies after allogeneic hematopoietic cell transplantation. It can eradicate minimal residual disease or be used to rescue a hematologic relapse, being able to induce durable remissions in a subset of patients. With the increased use of haploidentical hematopoietic cell transplantation, there is renewed interest in the use of donor lymphocytes to either treat or prevent disease relapse post transplant. Published retrospective and small prospective studies have shown encouraging results with therapeutic donor lymphocyte infusion in different haploidentical transplantation platforms. In this consensus paper, finalized on behalf of the Acute Leukemia Working Party of the European Society for Blood and Marrow Transplantation, we summarize the available evidence on the use of donor lymphocyte infusion from haploidentical donor, and provide recommendations on its therapeutic, pre-emptive and prophylactic use in clinical practice.

Some recommendations of Infectious Diseases Society of America for diagnosis and treatment of invasive aspergillosis with lung involvement, which is the most frequent clinical manifestation of aspergillar infection.Full version of recommendations was published in one of the editions of journal “Clinical microbiology and antibacterial chemotherapy” (2008. Т. 10. №2). Fungi of Aspergillus kin is the main etiologic cause of life-threatening infections in immunocompromised patients, such as patients with prolonged neutropenia, advanced stages of HIV infection, hereditary immunodeficiency states and after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and/or lung transplantation. This version of recommendations for treatment of invasive aspergillosis from IDSA (Infectious Diseases Society of America) substituted the former recommendations published in 2000. The level of evidence of treatment data is given in according with standard scheme used in the IDSA guidance.

Prognosis of patients with acute myeloid leukemia receiving intensive chemotherapy is defined as kariotyping anomalies, and mutations of genes responsible for surviving and self-maintenance leukemic cells. In the giving work incidence of mutation involving genes FLT3, c-KIT, NPM1 having prognostic value in adults is analysed. Kynase domain FLT3 mutation in 18 from 83 patients (21.6%) was detected. 2 from 18 patients (11%) achieved complete remission, 5 from 18 patients (27.7%) with FLT3 mutations were refractory to therapy, two patients died from treatment complications, 9 from 18 (50%) relapsed. Only 2 patients (11%) are alive in continuous complete remission. с-KIT mutations detected in 14.2% (2/14 patients), and NPM1 mutations represented allelic polymorphism. Thus, kynase domain FLT3 mutation are prognostic unfavorable.

Therapy results of 61 patients (children and adolescents, 2—17 years of age; a median — 8.76 years) with B-cells non-Hodgkin lymphomas III—IV stage and B-cell acute leukemia (B-NHL/B-ALL), treating according to protocol with polichemotherapy and monoclonal antibodies against B-cell receptor CD20 — rituximab (Mabthera, Hoffmann La Roche) applications, are presented. The protocol consist of cytoreductive phase and 6 polychemotherapy courses, similar to original protocol B-NHL-BFM 90, with exception of decreasing methotrexate dosage in 2 first courses from 5 to 1 g/m2/24 hours. Rituximab — 375 mg/m2 IV — in Day 0 first 4 courses was given.Four patients have died within first 3 weeks of therapy, 2 were primary refractory patients and 55 patients have achieved complete remissions. There was no case of remission death; relapse has developed only at one patient (neuroleukemia in patient with Burkitt lymphoma). 54 patients (88%) are in continuous complete remission. Overall and event-free survival rate were 0,90±0,04 with median follow-up after therapy ending 34 months (12—53 months). Authors discuss tactics of such combined therapy, features of remission registration, considering that at third of patients, residual mass was long-term detected, and for which assessment besides CT/MRT a positron-emission tomography (PET) and/or a repeated biopsy were done. Also it is possible to conclude that with use of target therapy chemotherapy intensity of such tumors can be reduced. Authors concluded that in cases of target therapy efficacy criteria should be modified, and it is necessary to use as much as possible informative technique to remission control (PET, minimal residual disease assessment with use of molecular-genetic methods).

Epstein–Barr virus (EBV) is a member of herpes-viruses family. Now it is well-known, that it can be a cause of wide spectrum lymphoproliferative disoders. The given problem is especially serious after hematopoietic stem cells transplantation (HSCT); frequency of death linked to this complication can achieve 50–90 %. Risk factors for development of posttransplant lymphoproliferative syndromes (PTLS) are: using partially compatible graft, T-cell depletion, presence of severe acute graft-versus-host reaction (GVHD), using of antitymocytic globulin, etc. Severity of associated with EBV posttransplant complications and problems relate to absence of clear diagnostic algorithm, PTLS prophylactic and therapy schedule are argument for necessity of the further studies.

New guidelines on diagnosis and treatment of a primary immune thrombocytopenia (ITP) in children and adults, on the basis of modern pathogenesis conceptions and considering possibilities of hematological practice in Russia are presented. Guidelines are based on a consensus of international and Russian experts on ITP. Diagnostic features of ITP are noted and various I, II and III lines treatment, including new effective medical products which have recently appeared in Russia, in particular, trombopoietin receptor agonists (romiplostim, eltrombopag), considerably expanded conservative treatment possibilities are offered. The presented guidelines can help hematologists to develop individual schedule for each given patients with ITP and to improve patients quality of life.

Various conditioning regimes influence on sexual development and gonads function in children and adolescent after stem cell transplantation was studied. 45 patients were enrolled in the study: 23 patients with aplastic anemia (АА), who received non-myeloablative conditioning, and 22 patients with acute myeloid leukemia (AML) after myeloablative conditioning. Sexual development was estimated by Tanner classification, laboratory tests included gonadotropic hormones, sex steroids, antiMullerian hormone and inhibin B level detection. It has been shown that non-myeloablative conditioning does not influence sexual development in any patients. Extremely high frequency of gonads lesion in patients received myeloablative conditioning was registered: hypergonadotrophic hypogonadism in 8 from 12 girls with severe decrease of ovaries functional reserve in all patients; germinal epithelium lesion in 9 from 10 boys.

Early complications of hematopoietic stem cell transplantation (HSCT) of vascular origin within the first 30-60 days after HSCT, which are resulting from endothelial injury, are an important cause of transplant-associated morbidity and mortality. Authors describe a case report serving as an example of several endothelial syndromes development with overlapping clinical features. Risk factors and therapy approaches of these complications are discussed.

Cytomegalovirus infection (CMV) is an extremely serious problem in patients after hematopoietic stem cells transplantation (HSCT). We have evaluated importance of major risk factors CMV development in patients after allogeneic HSCT (n = 168) from related (n = 56), unrelated (n = 90) or haploidentical donors (n = 22). Clinical importance of HSCT type as risk factors CMV development was shown; patients after unrelated or haploidentical HSCT had the worst prognosis. We also demonstrated that ≥ 2 grade acute GVHD statistically significant increase CMV reactivation probability after any types of HSCT. Comparison of infection reactivation frequency and event-free survival between subgroups of patients, received HSCT from CMV-positive and CMV-negative donors, absence of any differences was revealed. In the second group CMV infection was more severe. Preventive gancyclovir treatment has not shown efficacy and any influence on frequency of CMV reactivation.

Chronic immune thrombocytopenic purpura (ITP) is one of the most frequent immune hematological disorders in which development the leading part is played by the antibodies directed against a narrow spectrum of platelet antigens. Though death rate at chronic ITP is insignificant, no more than 1%; disease strongly reduces patients quality of a life, and frequently requires difficult and expensive treatment. Antiplatelets antibodies mediating ITP, are directed against glycoprotein GP IIb/IIIa and, less often, GP Ib/IX. At ITP can be registered both raised bone marrow platelet production and accelerated peripheral blood platelet clearance, and strongly decreased production and normal platelet life-span. Antibodies to platelet membrane glycoproteins directly disturb platelet production in bone marrow. Standard drug interventions at chronic ITP - glucocorticoids in various doses, high doses intravenous immunoglobulin, anti-D immunoglobulin, rituximab - rare lead to stable increase platelet number while in 2/3 patients splenectomy is effective. The new therapy approach in chronic ITP is stimulation of platelet production in bone marrow. Recombinant fusion protein Romiplostim stimulates trombopoietin receptor and it is effective at long-term therapy in 87% patients with chronic ITP irrespective of previous splenectomy. TPO antibodies development with romiplostim therapy was not revealed.

Background: The advent of next-generation sequencing (NGS) has revolutionized the analysis of genetic data, enabling rapid identification of pathogenic variants in patients with inborn errors of immunity (IEI). Sometimes, the use of NGS-based technologies is associated with challenges in the evaluation of the clinical significance of novel genetic variants. Methods: In silico prediction tools, such as SpliceAI neural network, are often used as a first-tier approach for the primary examination of genetic variants of uncertain clinical significance. Such tools allow us to parse through genetic data and emphasize potential splice-altering variants. Further variant assessment requires precise RNA assessment by agarose gel electrophoresis and/or cDNA Sanger sequencing. Results: We found two novel heterozygous variants in the coding region of the LYST gene (c.10104G>T, c.10894A>G) in an individual with a typical clinical presentation of Chediak–Higashi syndrome (CHS). The SpliceAI neural network predicted both variants as probably splice-altering. cDNA assessment by agarose gel electrophoresis revealed the presence of abnormally shortened splicing products in each variant's case, and cDNA Sanger sequencing demonstrated that c.10104G>T and c.10894A>G substitutions resulted in a shortening of the 44 and 49 exons by 41 and 47 bp, respectively. Both mutations probably lead to a frameshift and the formation of a premature termination codon. This, in turn, may disrupt the structure and/or function of the LYST protein. Conclusions: We identified two novel variants in the LYST gene, predicted to be deleterious by the SpliceAI neural network. Agarose gel cDNA electrophoresis and cDNA Sanger sequencing allowed us to verify inappropriate splicing patterns and establish these variants as disease-causing. [ABSTRACT FROM AUTHOR]

Survival after allogeneic hematopoietic stem cell transplantation (allo-HSCT) for severe idiopathic aplastic anemia (SAA) has improved in recent years, approaching 75% at 5 years. However, an SAA-adapted composite endpoint, graft-versus-host disease (GvHD) and relapse/rejection-free survival (GRFS), may more accurately assess patient outcomes beyond survival. We analyzed GRFS to identify risk factors and specific causes of GRFS failure. Our retrospective analysis from the Severe Aplastic Anemia Working Party of the European Society for Blood and Marrow Transplantation included 479 patients with idiopathic SAA who underwent allo-HSCT in two conventional situations: i) upfront allo-HSCT from a matched related donor (MRD) (upfront cohort), and ii) allo-HSCT for relapsed or refractory SAA (rel/ref cohort). Relevant events for GRFS calculation included graft failure, grade 3-4 acute GvHD, extensive chronic GvHD, and death. In the upfront cohort (n=209), 5-year GRFS was 77%. Late allo-HSCT (i.e., >6 months after SAA diagnosis) was the main poor prognostic factor, specifically increasing the risk of death as the cause of GRFS failure (hazard ratio [HR]=4.08; 95% confidence interval [CI]: 1.41-11.83; P=0.010). In the rel/ref cohort (n=270), 5-year GRFS was 61%. Age was the main factor significantly increasing the risk of death (HR=1.04; 95% CI: 1.02-1.06; P

Background. Extramedullary infiltration (EI) is relatively common in children with acute myeloid leukemia (AML) (up to 20-25 %). However, its clinical and prognostic significance remains poorly understood.Objective: to describe clinical features and to define prognostic significance of EI in children with AML.Materials and methods. The subjects of retrospective observational study were 228 children with de novo AML. The median age was 6.6 years. All of them were treated according to the protocol AML-MM-2006 from April 2007 to June 2018.All patients with EI were divided into three cohorts according to the localization of the lesions: 1) central nervous system (CNS) involvement (CNS group), 2) other localizations apart from CNS (myelosarcomas (MS) group), 3) combined lesions (CNS + MS group).Results. EI was diagnosed in 84 patients (36.84 %) with de novo AML. Among them 47 (55.95 %) had CNS involvement, 20 (23.81 %) had MS, 15 (17.86 %) had both CNS involvement and MS. 5-year overall survival (OS) rate was slightly higher in patients with CNS involvement than in children without EI - 80 ± 12 % vs 71 ± 9 %, p = 0.26, however OS in patients with MS was significantly lower - 45 ± 16 % vs 71 ± 9 %, p