Background. Nowadays, hematology is a dynamically developing science due to the in-depth study of the molecular mechanisms of a particular disease. A better understanding of oncohematological diseases biology makes it possible to synthesize new targeted drugs, which have a favorable therapeutic effect. In particular, in multiple myeloma, after the introduction of proteasome inhibitors and immunomodulatory drugs into clinical practice, an improvement in overall survival was observed. However, characteristics of the mechanisms of transformation normal plasma cells into malignant ones are still difficult; therefore, the study of the pathobiological basis of multiple myeloma is currently an urgent task.The objective: to evaluate the possible influence of MAGE-C1 gene expression and the presence of mage-c1 protein in patients with newly diagnosed multiple myeloma on the anti-tumor response after bortezomib-containing therapy.Materials and methods. A prospective study included 33 multiple myeloma patients. The diagnosis was established according to International Myeloma Working Group criteria (IMWG, 2014). In 32 patients the induction therapy included bortezomib-containing courses, in one patient lenalidomide was included in the first-line regimens. The MAGE-C1 gene expression by real-time polymerase chain reaction and mage-c1 protein by immunohistochemistry in plasma cells bone marrow, were determined for all patients at the debut of multiple myeloma. As a control group was examined the bone marrow material of healthy donors.Results. When assessment the statistical relationship between the expression of MAGE-C1 gene and mage-c1 protein, it was found that there was no high expression of mage-c1 protein at low values of MAGE-C1 gene expression. At the same time, high expression of the gene was always associated with protein expression above normal values. The analysis aimed at finding the relationship between MAGE-C1 gene and mage-c1 protein detection and the degree of antitumor response after 6 courses of induction therapy showed that high expression of the studied parameters was associated with a worse response to bortezomib-containing treatment.Conclusion. We confirmed that the results of the two methods were comparable. Single factor analysis showed that patients with decreased MAGE-C1 gene and mage-c1 protein expression levels achieved a significantly higher antitumor response to bortezomib-containing regimens, while high expression was accompanied by refractoriness to bortezomib.

Background: Background: Multiple myeloma (MM) course is often complicated by extramedullary lesions - plasmacytomas, which pathogenesis is still not completely understood. Genome instability underlies dissemination in various neoplasias, including MM. An important manifestation of genetic instability is the loss of heterozygosity, i.e., the loss of the second allele in the regions of the genome featuring inherited polymorphisms. STR profiling is a method that allows simultaneously assessing the degree of DNA degradation and giving an integral assessment of the stability of the tumor genome. To assess the heterogeneity of MM, not only the tumor substrate in the bone marrow and plasmacytoma but also in circulating cell-free DNA (cfDNA) present in the peripheral blood plasma could be effectively analyzed. Methods: Methods: 11 of bone 6 plasmacytoma GeneMapper v.4 software (Applied Biosystems,

Background. Multiple myeloma (MM) is a hematological malignancy with plasma cells as substrate. Sometimes MM is characterized by plasmacytomas, i.e., intra- and extraosse-ous tumors. A paraffin block containing plasmacytoma substrate provides valuable material to be used for analyzing the molecular biological characteristics of tumor. STR-profil-ing is a method for simultaneous evaluation of DNA degradation and integral assessment of tumor genome stability. Aim. To describe STR-profiles of plasmacytoma DNA isolated from archival samples and to assess the integral stability of tumor genome against control DNA of patients. Materials & Methods. The retrospective study enrolled 10 MM patients with plasmacytoma (7 women and 3 men) aged 34-62 years (median 53.5 years) who were treated at the National Research Center for Hematology from 2013 to 2021. Paired tumor/control DNA samples were obtained from all 10 patients. Results. The present paper takes the first step in attempting a large-scale molecular genetic study of MM and provides first findings on the loss of heterozygosity (LOH) in plasmacytoma genome. All 10 patients showed LOH variants with different allelic loads having either deletion/ quantitatively neutral LOH or duplication of one of the two alleles and involving 1-8 STR-loci. In plasmacytoma substrate the number of loci with LOH tended to be higher in the group with MM relapses compared with plasmacytomas identified at disease onset. According to the data analysis, LOH was frequently (in 4 out of 10 cases) detected on chromosomes 1 (1q42), 6 (6q14), 7 (7q21.11), 13 (13q31.1), and 21 (21q21.1). Conclusion. The present paper shows the effectiveness of molecular analysis of DNAs being isolated from complex archival material consisting of paraffin blocks with plasmacytomas.

Background: Malignant plasma cells (PC) immunophenotype is well understood and represented by a large group of antigens expressed with varying frequency. Clusters of differentiation are both diagnostic and prognostic factors. Some of these serve as targets for monoclonal antibody and genetically engineered T-cells with chimeric antigen receptors in multiple myeloma (MM) patients. These molecules are SLAMF7 (Signaling lymphocytic activation molecule factor - CD319) and BCMA (B-cell maturation antigen - CD269). Membrane subunits of these molecules can be cleaved by enzymes (f.ex. γ-secretase) and released to the plasma as soluble proteins and can be easily detected using readily available commercial enzyme-linked immunosorbent assays (ELISA). SLAM family of receptors is a self-regulating and stimulates tumor proliferation. Therefore, soluble BCMA detection is one of the predictors of refractory disease. Aims: To compare the CD269, CD319 expression on malignant PCs and sBCMA, sSLAMF7 protein concentrations in serum of newly diagnosed multiple myeloma patients. Methods: The prospective study included 18 patients (8 women, 10 men) aged 26 to 70 years (median 54) with NDMM between June and September 2021. They received VRD induction therapy (2-4 cycles, median 4). Malignant PCs immunophenotyping was determined by multicolor flow cytometry (MFC) (CD319; CD269; CD138; CD38; CD19; CD45; CD81; CD27; CD20; CD56; CD200; CD117). Concentration of soluble BCMA, SLAMF7 in serum was detected using commercial ELISA (RayBio® Human CRACC/SLAM7 ELISA Kit, RayBio® Human TNFRSF17 ELISA Kit). Statistical analyses were performed using SPSS. Results: The results of the CD269, CD319 and sSLAMF7, sBCMA protein concentrations in the serum of NDMM patients are presents in Table 1. According to MFC of bone marrow malignant PCs, the presence of CD319 is detected in 90% of patients (n=16). sSLAMF7 protein is detected in the serum of 39% of patients (n=7) in the range of 1.14 to 5.43 ng/ml (median 2.9), in 61% of cases (n=11) sSLAMF7 wasn’t detected. In 56% of cases sSLAMF7 wasn’t detected, but CD319 was detected on the surface of malignant PCs. No cases of sSLAMF7 without the presence of CD319 on the malignant PC were detected in this study. CD269 was detected on malignant PCs in 45% of patients (n=8) and in 55% of cases (n=10) malignant PCs was CD269 negative. sBCMA was detected in 100% of patients (n=18). The concentration ranged from 122.33 to 11617.32 pg/ml (median 2139.7). In the group of CD269 positive patients, sBCMA concentration was significantly (p=0.001) higher than in CD269 negative group: medians were 5334.12pg/ml (1933.6-11617.3) vs 318.87pg/ml (122.3-3962.2), respectively. After induction therapy response was complete response in 16.5% (n=3), very good partial response in 12% (n=2), partial response in 66% (n=12), and progression disease in 5.5% (n=1). However, statistically significant correlations were not found between the depth of response, the presence of CD319, CD269 on the malignant PCs or/and the concentration of sSLAMF7, and sBCMA. Image:Summary/Conclusion: The high frequency of CD319-positive patients was not accompanied by detection of sSLAMF7 in 61% of patients. Whereas, 100% sBCMA detection in NDMM patients was not followed by CD269 detection on malignant PC in half of the patients. These findings may indicate both the heterogeneity of MM and the necessity for further studies of sBCMA and sSLAMF7 to determine their prognostic significance.

Background: Bone plasmacytomas forms as a result of destruction of bone structures by tumor proliferate and its escape out of the medullar cavity. According published data, the incidence of bone plasmacytomas at the time of plasma cell myeloma (PCM) diagnosis ranges from 7 to 32,5%, whereas rates are higher in a relapse. Bone plasmacytomas in most cases are composed of mature plasma cells, while immature morphology is mainly characteristic for extramedullary plasmacytomas. It is shown, that plasmacytoma cells are the descendants of bone marrow tumor plasma cells. Due to acquisition of new qualities as a downregulation of adhesion molecules, secretion of growth factors, activation of oncogenes and antiapoptotic factors, new subclones of tumor cells can survive without support from bone marrow microenvironment. Aims: to compare morphological consistency and immunohistochemical characteristics of PCM substrate in bone marrow trephines and bone plasmacytomas biopsy specimens. Methods: In retrospective study were included 13 patients with PCM aged from 36 to 67 years (median 55 years). At the time of diagnosis all patients underwent biopsy of bone plasmacytoma. The diagnosis of PCM verified according to IMWG criteria (2014). All patients have been examined with routine laboratory tests, immunochemical analysis of serum proteins and urine, cytologic examination of bone marrow smears, histologic and immunohistochemical examinations of bone marrow trephines and bone plasmacytomas biopsy specimens, instrumental examinations (CT, MRI). Immunohistochemical staining of bone marrow trephines and bone plasmacytomas biopsy specimens made with using a stainer Leica Bond-Max, reactions with antibodies to CCND1 (clone SPA4, Cell Marque), NSD2 (clone 29D1, Abcam), c-MAF (clone ERP16484, Abcam). The cut-off scores for determining positivity for markers have been taken 5% for CCND1, 10% for NSD2 and c-MAF (by T. Murase et al., 2019). Microscopic examination of histologic and immunohistochemical slides have been done on light microscope Leica 3000. Results: PCM substrate by histologic examination in bone marrow trepnines and bone plasmacytomas biopsy specimens were presented by mature plasma cells. In two patients (1 and 8) in bone plasmacytoma have been seen proplasmocytes between mature plasma cells, but in bone marrow have been seen only clusters of mature plasma cells. According to the results of immunohistochemical examination the expression of proteins-products of oncogenes have been revealed in most patients (12 of 13) in plasma cells of bone marrow and bone plasmacytomas, wherein in half of the patients (patients 7-12) the expression of proteins-products of oncogenes coincides in bone marrow and bone plasmacytoma. We have revealed a discordant expression in another patients (tabl. 1). We’ve paid a special attention to the patients 1-3, in whom in bone marrow have been revealed an expression of 1/3 protein, but in bone plasmacytoma – 2/3 (patients 2 and 3) or all of 3 studied proteins (patient 1). This phenomenon is possible to consider as a heterogeneity of clonal composition of bone plasmacytoma as a consequence of tumor evolution. Image:Summary/Conclusion: Discordant expression of proteins-products of oncogenes NSD2, cyclin D1 and c-Maf in tumor cells in bone marrow and bone plasmacytoma could be evidence of different tumor clones activation in PCM.

Background: Transplantation of autologous blood stem cells (auto-HSCT) is one of the key methods in multiple myeloma (MM) treatment recommended to patients under 65 years of age and good somatic status. The early post-transplantation period is associated with the development of deep myelotoxic agranulocytosis, disruption of mucous membranes of gastrointestinal tract and high risk of infectious complications. Aims: Determine the incidence of infectious complications and the spectrum of pathogens in MM patients after autologous transplantation. Methods: Our study includes 111 MM patients (51 male) aged 19 to 65 (median age 53) who performed an auto-HSCT between 2018 and 2020. In 99 patients Durie-Salmon stage was considered as III, in 1 - as II and in 11 - as I. Myeloma nephropathy was observed in 23 patients, programmed hemodialysis in 4. Prior to the auto-HSCT 42 patients had reached the CR, 48 - VGPR, 15 - PR, 4 - stabilisation, 2 - progression. Hematopoietic stem cells were mobilized and 2,1-23,15x106/kg CD34+ cells (median 7,7) collected. Conditioning was performed with melphalan 200mg/m2 (n=93) and 140 mg/m2 (n=15), due to myeloma nephropathy and hemodialysis. Agranulocytosis lasted from 6 to 97 days (median 12). The colonization of non-sterile compartment was investigated before and in agranulocytosis. In case of the febrile fever, Bactec blood culture systems and chest CT were performed, in case of the oral mucositis - microbiological mucus examination from tonsils and the throat back wall. If pneumonia was established by CT, fibrobronchoscopy was performed with microbiological, molecular-biological and PCR examinations of bronchoalveolar lavage (BAL). Viral blood profile was investigated by PCR for CMV, HHV-6, Herpes simplex viruses. During neutropenia all patients were given antiviral prophylaxis by valaciclovir. Due to infection developing, broad-spectrum antibiotics were prescribed. Results: Infectious complications in the early post-transplant period was noted in 94.6% of MM patients. The most frequent were oral mucositis (95.5%) and necrotic enteritis (85.6%). Microbiological analysis of non-sterile compartment in 101 patients revealed 52 cases of E.coli without ESBL, 35 - with E.coli ESBL. In agranulocytosis, 93 cases were detected: E.coli without ESBL (n=32), E.coli ESBL (n=19). Bacteriemia was identified in 18 patients: 7 cases of gram-positive and 11 of gram-negative. Pneumonia, confirmed by chest CT, was detected in 17 (15.3%) patients, and in 8 cases fibrobronchoscopy was performed. The pathogens etiology was established in 5 patients - gram-positive (n=3) and gram-negative (n=2). Viral infection associated with HHV-6 was found in 8%. Analysing non-sterile compartment before the therapy and in case of bloodstream infection, they matched in 7 cases, a difference was found in 4 cases (C.koseri without ESBL, P.aeruginosa, C.putigena). In 110 patients all infections were successfully cured and patients were discharged satisfactorily. Only one patient died at +9 day of auto-HSCT from bilateral pneumonia caused by S. aureus and bacteriemia associated with P. aeruginosa and S. aureus. Image:Summary/Conclusion: In our study, the most frequent infectious complications in patients with MM in the early post-transplantation period, were oral mucositis and necrotic enteritis, mainly I-II degree of severity. Complications, such as pneumonia and bacteriaemia, developed in 32%. Viral infections were detected in 8% of patients and associated with HHV6. Early post-transplant mortality was 0.9%.

Актуальность. Множественная миелома (ММ) — злокачественная опухоль системы крови, субстратом которой являются плазматические клетки. Иногда при ММ отмечается появление костных и внекостных опухолей — плазмоцитом. Парафиновый блок, содержащий субстрат плазмоцитомы, — ценный материал, который можно использовать для изучения молекулярно-биологических характеристик опухоли. Методом, позволяющим одновременно оценить степень деградации ДНК и дать интегральную оценку стабильности генома опухоли, является STR-профилирование. Цель. Дать характеристику STR-профилей ДНК плазмоцитом, выделенной из архивных образцов, оценить интегральную стабильность генома опухоли относительно контрольной ДНК пациентов. Материалы и методы. В ретроспективное исследование включено 10 пациентов с ММ, осложненной плазмоцитомой (7 женщин, 3 мужчины) в возрасте 34–62 года (медиана 53,5 года), которые находились на лечении в ФГБУ «НМИЦ гематологии» МЗ РФ с 2013 по 2021 г. У всех 10 пациентов удалось выделить парные образцы ДНК опухоли/контроля. Результаты. В работе сделан первый шаг к масштабному молекулярно-генетическому исследованию ММ, собраны первые данные о потере гетерозиготности (ПГ) в геноме плазмоцитом. У всех 10 пациентов выявлены варианты ПГ с разной аллельной нагрузкой, которые предполагают делецию/количественно нейтральную ПГ либо дупликацию одного из двух аллелей, затрагивающие 1–8 STR-локусов. Наблюдалась тенденция к большему числу локусов с ПГ в субстрате плазмоцитом из группы с рецидивами ММ по сравнению с плазмоцитомами, выявленными в дебюте заболевания. При анализе данных отмечена часто встречающаяся ПГ (4 из 10 случаев) на следующих хромосомах: 1 (1q42), 6 (6q14), 7 (7q21.11), 13 (13q31.1), 21 (21q21.1). Заключение. Представленная работа демонстрирует эффективность молекулярного анализа ДНК, выделенной из сложного архивного материала парафиновых блоков плазмоцитом.

Background. Multiple myeloma complicated by extramedullary plasmacytoma is an unfavorable variant of the disease. It remains unknown what triggers tumor transformation. The review presents literature data on the pathogenesis of extramedullary disease, as well as a clinical example of a comprehensive study of the tumor substrate.Aim. To study the molecular and biological characteristics of the tumor substrate of the bone marrow and extramedullary plasmacytoma using various research methods.Materials and methods. A 55-year-old patient was admitted to National Medical Research Center for Hematology with a diagnosis of multiple myeloma occurring with extramedullary plasmacytoma of the retroperitoneal space. dNA was isolated from samples of different localization (blood plasma, Cd138+ bone marrow cells, plasmacytoma and buccal epithelial cells). The profile of short tandem dNA repeats (STR) from the obtained samples was studied by multiplex polymerase chain reaction followed by fragment analysis. fluorescent in situ hybridization (fISH) of bone marrow Cd138+ cells was performed using various dNA probes. Comparative genomic hybridization on a microarray (arrayCGH) plasmacytoma dNA was also performed. The mutation profile of the KRAS, NRAS, BRAF genes was studied by Sanger sequencing in tumor samples of various localizations.Results. The induction therapy (vCd (bortezomib + cyclophosphamide + dexamethasone), vRd (bortezomib + lenalidomide + dexamethasone), daratumumab therapy) was ineffective, death occurred 4 months after the first clinical manifestations appeared. Comparison of STR markers of circulating cell-free tumor dNA (cfdNA), Cd138+ bone marrow cells, and plasmacytoma revealed the largest number of involved loci exactly in plasmacytoma’ dNA. A mutation in the NRAS gene was found only in plasmacytoma’ dNA. This indicates the presence of another clone of tumor cells in the extra-medullary plasmacytoma. Molecular karyotyping of plasmacytoma using the arrayCGH method revealed rearrangements of many chromosomes. 1p32.3 bi-allelic deletion, amplification of 1q21, 8q24/MyC rearrangements and del17p13 were confirmed by arrayCGH molecular karyotyping and fISH studies in bone marrow and plasmacytoma.Conclusion. A comprehensive molecular genetic study of the extramedullary plasmacytoma’ substrate is necessary to understand the pathogenesis mechanisms and, on this basis, to develop differentiated therapeutic approaches.

Introduction. Autologous haematopoietic stem cell transplantation (auto-HSCT) is a highly effective treatment for multiple myeloma (MM). Auto-HSCT allows a signifi cant improvement of haematological response leading to higher overall survival and quality of life in MM patients. Nonetheless, the majority of patients develop relapse.Aim — a comparison of clinical MM relapses developing at variant terms after auto-HSCT.Patients and methods. A retrospective study enrolled 65 MM patients aged between 39 and 64 years. All patients had auto-HSCT during 2009–2019, all had achieved complete response (CR) or very good partial response (VGPR) and all since developed immunochemical MM relapse in laboratory evidence. Patients were divided in two cohorts by relapse term, the early (within 12 months of auto-HSCT) and late relapse.Results. Early immunochemical relapse was diagnosed in 13 (20 %), late relapse — in 52 (80 %) patients. The dependence between relapse term and depth of post-auto-HSCT antitumour response has been determined. The proportion of CR patients was signifi cantly higher in late than in early relapse (55.8 vs. 23 %). In follow-up, 60 patients (92.3 %) were initiated on antirelapse therapy, all early relapse and 90.3 % late relapse patients. On day +100 of auto-HSCT, CR patients had later relapse vs. VGPR individuals (median 24 vs. 19.9 months, p = 0.08) with signifi cantly weaker paraprotein secretion resembling the clinical course of monoclonal gammopathy of unclear signifi cance (MGUS).Conclusion. Auto-HSCT allows long-term control of the disease. A signifi cant prognostic factor is antitumour response on +100 day of auto-HSCT. Patients attaining CR have later relapse progressing in a MGUS-like manner. Patients with late indolent relapse can be managed long-term without antitumour therapy.

Objective of the study: analysis of AL-amyloidosis (AL-A) diagnostics in real clinical practice and to determine the main approaches for the earlier detection of this disease.Materials and methods. A retrospective analysis of medical records of 34 patients with newly diagnosed AL-A.Results. The median time from first symptoms appearance to the diagnosis was more than 2 years. Most often, the pathological process in AL-A involves the kidneys, heart and gastrointestinal tract; moreover, at the time of diagnosis, most patients already have an injury of 2 or more organs. In half of the patients, a biopsy of the damaging organ was performed to verify the diagnosis; according to our data, histological examination of “easily accessible” locus not less informative. Symptoms characteristic of amyloidosis, such as periorbital purpura or macroglossia, are observed in a small part of patients and refer to late manifestations of the disease.Conclusion. The versatility of AL-A manifestations leads to a late diagnosis, which affects overall survival. The main signs were highlighted that should alert doctors in relation to this pathology.

Introduction. Cytogenetic and genomic traits of tumour cells are considered the key mediating factors in multiple myeloma (MM). Selected chromosomal abnormalities are prognostic of therapeutic response and patient survival in MM.Aim — to assess of the diversity and rate of chromosomal abnormalities in MM patients and their association with the disease course.Materials and methods. The study enrolled 134 MM patients with pre-treatment bone marrow FISH assay screening for chromosomal abnormalities: t(11;14), t(4;14), t(14;16), t(14;20), t(6;14), hyperdiploidy, del13q14/-13, del17p13/TP53, amp1q21, t(8q24)/cMYC. The studied criteria at the MM onset were: hemogram, lactate dehydrogenase (LDH) activity, calcium, β2-microglobulin and creatinine concentrations, punctate cytology, bone marrow trephine biopsy and/or soft tissue biopsy histology, bone X-ray, immunochemical variant of MM, disease staging. A median follow-up was 20 months (3.2–77.4).Results. The primary chromosomal abnormality rate was 82.9 %, among them t(14q32)/IGH — 29.1 %, multiple trisomies — 46.3 % and their combination — 7.5 %. The rates of particular t(14q32)/IGH): t(11;14) — 16.4 %, t(4;14) — 12.7 %, t(14;16) and t(14;20) — 3.7 and 2.2 %, respectively. The secondary chromosomal abnormality rate was 69.4 %, among them del13q14/-13 — 40.3 %, amp1q21 — 39.6 %, t(8q24)/cMYC — 17.2 %, del17p13/TP53 — 12.7 %, del1p32 — 2.2 %. Analyses of the primary–secondary abnormality combinations showed that del13q14/-13 is more frequently combined with t(4;14) and less frequently with trisomies (p < 0.05). Amp1q21 occurs more frequently with t(4;14) and less — with t(11;14) (p<0.05). Patients with t(4;14) more frequently (p < 0.05) had anemia at a hemoglobin level<100 g/L, and the presence of amp1q21 and del17p13/TP53-enhanced serum LDH activity (p < 0.05). Abnormality t(8q24)/cMYC more often co-occurred with higher serum β2-microglobulin concentrations (p < 0.05). A three-year overall survival (OS) in del17p13/TP53-positive patients was 35.5 vs. 71.3 % in the negative (p = 0.002) and 50.8 vs. 67 % — in t(8q24)/cMYC-positive and negative patients, respectively (p = 0.001). Patients without amp1q21, with one, with two or more additional 1q21 copies had a five-year OS 79.4, 67.3 and 20.9 %, respectively (p = 0.0016), and a two-year progression-free survival (PFS) 83, 50 and 0 %, respectively (p = 0.005).Conclusion. We establish a negative impact of del17p13/TP53 and t(8q24)/cMYC on patients’ OS in MM, as well as unfavourable effect of amp1q21 on OS and PFS in the presence of two or more additional copies of 1q21 loci.

AIM:To assess the safety and efficacy of autologous haematopoietic stem cell transplantation (auto-HSCT) in multiple myeloma (MM) patients with dialysis-dependent renal failure.MATERIALS AND METHODS:During a period from May 2010 to December 2016 fourteen MM patients with dialysis-dependent renal failure aged 48 to 65 years underwent auto-HSCT. After the induction therapy complete response, very good partial response, partial response were documented in 64, 29, 7% of patients, respectively. In no case was a renal response achieved. Haematopoietic stem cell mobilization in most patients (13/14) was performed according to the scheme: G-CSF 10 g/kg. Melphalan in 3 dosages was used as pre-transplant conditioning: 100, 140 and 200 mg/m2; 13 patients underwent a single and in one case underwent a tandem auto-HSCT against the background of hemodialysis. Evaluation of the antitumor and renal response was assessed on the 100th day after auto-HSCT. Subsequently, against the background of programmed hemodialysis and in the setting of high-dosed melphalan (100200 mg/m2), 13 patients underwent a single and one patient underwent a tandem auto-HSCT. At +100 days after auto-HSCT, an antitumor response and renal response were assessed.RESULTS:The period of agranulocytosis after auto-HSCT was from 5 to 12 days (median 8,5) and was accompanied by infectious complications, cardiac and neurological dysfunctions. At +100 days after auto-HSCT, the complete response was confirmed in 71% patients and very good partial response was confirmed in 29% patients. The minimal renal response was registered in 2 patients (14%), hemodialysis was stopped. The transplant-related mortality was absent. After a median follow-up of 53 months 5-year progression-free survival was 59%, and overall survival was 93%.CONCLUSION:Carrying out auto-HSCT in patients with dialysis-dependent renal failure contributed to the achievement of a minimal renal response in 14% of cases, which allowed these patients to stop hemodialysis. Patients whose conditioning regimen was performed using melphalan at a dose of 200 mg/m2showed more frequent complications in the early post-transplant period compared to patients who received a lower dose of melphalan (100140 mg/m2). Auto-HSCT in MM patients with dialysis-dependent renal failure is a feasible and effective treatment method, which in some cases contributes to independence from hemodialysis.

Aim: to compare the results of tumor visualization when using 18F-FDG and 11C-methionine PET/CT after auto-HSCT in MM patients. Materials and methods. A prospective study included 27 MM patients subjected to 18F-FDG and 11C-methionine PET/CT on day 100 after auto-HSCT. Obtained images were visually and semi - quantitatively analyzed. Focal areas of increased uptake for every radiopharmaceutical agent (hypermetabolic foci) not associated with its physiological distribution were registered. Maximum Standardized Uptake Values (SUVmax) in pathological foci were automatically calculated for every radiopharmaceutical agent separately. PET/CT findings were compared to antitumor response achieved after auto-HSCT according to International MM Working Group criteria. Results. After auto-HSCT, the majority of patients (16/60%) achieved a complete response. Abnormal 18F-FDG uptake was registered in 37% (n=10) of patients, negative PET findings were obtained in 63% (n=17) of patients. 11C-methionine PET/CT revealed hypermetabolic foci in 67% (n=18) of patients, and there was no 11C-methionine uptake in 33% (n=9). Pathological foci of radiopharmaceutical agent uptake were 1.8 times more frequently revealed using PET/CT with 11C-methionine (p

Aim. To study immunohistochemical parameters of tumor plasma cells in bone marrow and to assess how the expression of adhesion molecule CD56 impacts overall survival (OS) of multiple myeloma (MM) patients. Materials & Methods. The trial included 35 patients (19 men and 16 women) aged 23 to 73 years (with median age of 58 years) with newly diagnosed MM. At disease onset plasmacytoma was diagnosed in 21 patients. In all patients bone marrow core biopsy was performed followed by histologic and immunohistochemical (IHC) examinations. IHC examination was based on the panel of CD56, CD166, CXCR4, Ki-67, and c-MYC/CD138 antibodies. Kaplan-Meier survival curves and significance assessment by means of Cox's F-Test were used. Results. Expression mean values of most of studied markers (CD56, CXCR4, c-MYC, and Ki-67) in bone marrow of patients without plasmacytoma (n = 14) appeared to be higher than in patients with plasmacytoma at MM onset. Expression mean value is understood as percentage ratio of plasma cells expressing a studied marker to total cell count of tumor substrate. High expression of chemo-kine receptors (CXCR4), and adhesion molecules (CD56) probably inhibits plasma cell migration and impedes ex-tramedullary tumor progression. Comparison of protein expression by tumor plasma cells in bone marrow in the groups with bone extramedullary plasmacytoma shows a distinct regularity referring to CD56 adhesion molecule. For example, CD56 expression is significantly (р < 0.05) lower in terms of the count of tumor plasma cells with marker expression in bone marrow of MM patients with extramedullary plasmacytoma compared with patients with bone plasmacytoma (1 ± 1 % vs. 65.71 ± 12.12 %). Comparison of MM patients' OS depending on CD56 expression by tumor plasma cells in bone marrow showed that 4-year OS of patients with CD56 expression in bone marrow was significantly higher being 80 % vs. 38 % in the group with CD56 expression less than in 10 % of tumor cells. Conclusion. Expression of adhesion molecule CD56 in tumor plasma cells in bone marrow can be regarded as a prognostic factor in MM. Probably, when at disease onset CD56 expression is identified in less than 10 % of tumor cells in bone marrow, more detailed additional examination of patients should be carried out to rule out extramedullary lesions in different organs and tissues.

Background: MRD-negative status in patients with MM after auto-HSCT is considered as a favorable prognostic factor. Despite the absence of aberrant plasma cells according to flow cytometry data on the 100th day after auto-HSCT, early recurrence of the disease is observed in some patients. Considering the biological features of MM, which consists in diffuse focal lesion of the bone marrow, the study of the bones of the entire skeleton with visual assessment methods are of scientific value. MRI, as the most sensitive method for determining the infiltrative changes in the bone marrow, may become one of the additional criteria for an antitumor response. Aims: To determine the effect of identified bone marrow lesions using MRI in MM patients who have MRD-negative status after auto-HSCT on progression-free survival (PFS). Methods 14 patients with MM (7 male and 7 female) aged 46 to 66 years (median age 56 years) were enrolled in a prospective study in the period from December 2015 to March 2017. The ISS stages at the time of diagnosis were as follows: stage I in 5 patients, stage II in 6, and stage III in 3 subjects. Soft tissue components were observed in 6 patients (43%). All patients received induction therapy with bortezomib. Following induction therapy, single auto-HSCT was performed in 14 patients. MRI of the spine and pelvic bones was carried out with a GE Signa Profile system without contrast enhancement 100 days after auto-HSCT. The nature of the lesions was determined and the bone marrow infiltration lesions (≥ 5 mm) counted. Statistical analysis was done using Statistica 10. Results: After auto-HSCT, the following antitumour response rates were observed: CR in 10 (72%), VGPR in 2 (14%) and PR in 2 subjects (14%). At 100 days after auto-HSCT all patients achieved strict CR (MRD-negativity according to flow cytometry). Focal changes in the bone marrow by MRI after auto-HSCT were detected in 12 patients: the minimum number of lesions was 1, the maximum was 40 (average 6). The distribution of patients according to the number of lesions after auto-HSCT was as follows: 0 - 1 in 7 (50%) patients, 2 - 40 in 7 (50%) cases. After auto-HSCT patients did not receive maintenance therapy they were monitored until the disease recurred. With a median follow-up of 21 months, relapse was documented among 8 patients. At the same time, in 3 of them, the results of MRI performed on the 100th day after auto-HSCT showed minimal bone marrow damage (less than 1 focus) and in 5 cases more than 1 lesion of the bone marrow was detected. 3-year PFS in the group of patients with minimal bone marrow damage on MR-tomograms (less than 1 lesion) was significantly (p <0.05) higher and amounted to 52% versus 18% in the group of patients with more than 1 lesion detected in the bone marrow by MRI data (Figure 1).Summary/Conclusion: Differences in PFS suggest that in patients with MRD-negativity after auto-HSCT, a whole-body MRI reveals focal lesions in the bone marrow. Flow cytometry examines only local bone marrow damage from a biopsy. MRI of the bone marrow can reduce the false-negative results of the determination of MRD after auto-HSCT.

Background . The study of influence of residual tumor mass, determined by magnetic resonance imaging (MRI), on the progression-free survival (PFS) remains an actual problem. Since the visual assessment of tumor bone marrow lesion can be one of the criteria for the subsequent personalized treatment choice in multiple myeloma patients.The objective of study was to determine the effect of bone marrow lesions detected by MRI after autologous hematopoietic stem cells transplantation (auto-HSCT) on PFS in multiple myeloma patients.Materials and methods . The prospective study included 60 patients who underwent spine and pelvic bones MRI on the 100 th day after autoHSCT.Results . Focal bone marrow changes were found in 47 of them – from 1 to 56 lesions (mean 6 ± 9). Significant (p = 0.01) differences of PFS in multiple myeloma patients depending on the presence or absence of tumor mass on 100 th day after auto-HSCT were revealed: with MRI negative status, 2-year PFS was 89 % versus 50 % in a group of patients with residual tumor mass.Conclusion . MRI-negative status after auto-HSCT is a favorable prognostic factor contributing to prolonged disease-free survival.

Background: The bone marrow microenvironment (BMME) is crucial for the development and persistence of multiple myeloma (MM). This specific BMME includes immune cells and soluble factors involved in the regulation of tumor pathogenesis and progression. Further, the success of immune-based therapies is likely conditioned by the specific characteristics of each patient́s immune system by means they should exert its beneficial effects. Aims: To characterize the cellular and soluble BMME in patients (pts) representing the complete spectrum of monoclonal gammopathies (MG) at diagnosis as well as in relapsed and refractory (RR) disease, to identify features potentially involved in disease pathogenesis. Methods: We have studied 163 samples from pts with MGUS (n = 9), high-risk smoldering MM (HRsMM) (n = 20), newly diagnosed MM (NDMM) (n = 26 £70 yo, n = 97>70 yo), and relapsed and refractory (RRMM) disease (n = 11), comparing the results with those from a cohort of age-matched healthy donors (HD; n = 26).We identified and quantified 25 BM immune populations (B, T and NK and subsets) using three 8-color combinations of MoAbs. We also analyzed the PB serum concentration of seven cytokines (IFNg, IL10, IL12p70, IL17a, IL4, IL6, MIP1a and TNFa) by Milliplex and complement levels (C3 and C4) and activity (CH50) in a subgroup of 14 MGUS and 10 NDMM cases. Statistics were performed using the IBM SPSS package v25. Results: First, we compared the results of the analysis of immune cell populations at diagnosis between samples from HD and MGUS, HRsMM and NDMM (Table 1). When compared to HD, an increased percentage of CD8+ T-cells was identified in MGUS pts, due to increased effector subsets. As per the HRsMM, higher percentages of all cell populations were identified: in CD4+ T-cells, based on the memory subsets; in CD8+, based on the effectors and in all NK subpopulations, except for the tissue resident subgroup. Interestingly, the same results were obtained when comparing HD vs NDMM cases. Then, we compared the results in the 3 pathogenic stages of the disease (MGUS vs HRsMM, MGUS vs NDMM and HRsMM vs NDMM) to identify potential changes associated with disease evolution. Again, MGUS differed from HRsMM by the increased percentage of CD4+ T-cells in the latter due to increased memory subsets but also in the higher percentage of NK cells due to the circulating and canonical subgroups. Similar results were identified when comparing MGUS and NDMM. Interestingly again, we did not find any significant difference when comparing HRsMM and NDMM cases. When we compared the characteristics of the BM immune ME between ND and RR cases, a decreased percentage in B cells and CD4+ T cells (based on reduced naïve subsets) was detected in the latter. Regarding the soluble factors, we found that, as compared to MGUS, in NDMM cases serum levels of IL17a were reduced but TNFa increased. C3 was above the reference ranges in 40% of MGUS vs. 73% of NDMM and C4 in 27% of MGUS vs. 64% of NDMM. In addition, CH50 was normal in all MGUS cases whereas it was reduced in 18% of NDMM pts.Summary/Conclusion: Analysis of the immune profile in pts with the complete spectrum of MG suggests significant and sequential changes associated with the transition throughout the different stages of the disease, both in cell populations and soluble factors. These results could be due to immune-vigilance in the precursor stages, which is lost with the progression and/or the treatment effect. Nevertheless, further analysis is needed to confirm these results and to ascertain their influence in MM development.