ISSN 1866-8836
Клеточная терапия и трансплантация

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Volume 8, Number 2
06/28/2019
Volume 8, Number 2
Editor-in-Chief
Afanasyev B. V. (St. Petersburg, Russia)
Co-Editors-in-Chief
Wagemaker G. (Rotterdam, Netherlands)
Zander A. R. (Hamburg, Germany)
Deputy Editor
Fehse B. (Hamburg, Germany)
Managing Editor
Chukhlovin A. B. (St. Petersburg, Russia)
Editorial Board
Aleynikova O. V. (Minsk, Republic of Belarus)
Borset M. (Trondheim, Norway)
Chechetkin A. V. (St. Petersburg, Russia)
Fibbe W. (Leiden, Netherlands)
Galibin O. V. (St. Petersburg, Russia)
Hölzer D. (Frankfurt a.M., Germany)
Klimko N. N. (St. Petersburg, Russia)
Kolb H.-J. (München, Germany)
Kröger N. (Hamburg, Germany)
Kulagin A. D. (St. Petersburg, Russia)
Lange C. (Hamburg, Germany)
Mamaev N. N. (St. Petersburg, Russia)
Mikhailova N. B. (St. Petersburg, Russia)
Moiseev I. S. (St. Petersburg, Russia)
Nagler A. (Tel-Aviv, Israel)
Nemkov A. S. (St. Petersburg, Russia)
Paramonov I. V. (Kirov, Russia)
Roumiantsev A. G. (Moscow, Russia)
Savchenko V. G. (Moscow, Russia)
Smirnov A. V. (St. Petersburg, Russia)
Uss A. L. (Minsk, Republic of Belarus)
Zubarovskaya L. S. (St. Petersburg, Russia)
In this Issue
The present CTT number starts with a review article by Dr. Elezov and Kudryavtsev concerns modern knowledge on expression and regulation of PD-1 receptor and its ligand PLD-1, like as on their role in diagnostics and prediction of cancer treatment with immune checkpoint inhibitors.

An original study by Dr. Popova et al. describes a representative cohort of patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT). The authors have tested the pre-transplant fungal infection in this group, and have found that prior invasive fungal disease did not produce a significant impact on transplant-related complications and overall survival.

A single-center study by Dr. T. Rudakova et al. presents original data on their experience with of thrombopoietin receptor agonists used in the patients with poor graft function after allo-HSCT. The workers have observed good clinical response in sufficient part of this group and safety of long-term administration of TPO receptor agonists.

The interdisciplinary study performed by Dr E. Kulagin et al. concerned pre-existing respiratory disorders as a possible factor of adverse effects upon posttransplant complications and survival of HSCT patients. The pulmonary comorbidities in children and adults before allo-HSCT proved to be a significant independent risk factor with respect to posttransplant disorders.

A comprehensive article by Prof. Mamaev et al. provides additional evidence on diagnostic utility of BAALC gene overexpression in acute myeloid leukemia. Combined overexpression of BAALC and WT1 genes may be used for regular post-transplant monitoring of relapses in this disorder.

A pilot study of mesenchymal stem cell infusions for treatment of ulcerative colitis by Prof. Galina Fedotovskikh et al. is focused, mainly, on morphological correlates of intestinal damage several months after standard and MSC-supplemented therapy. The article should elicit some discussions, due to small number of cases and the issues of morphological criteria used.

Two interesting clinical cases are presented by A. Smirnova et al. They illustrate a striking clinical effect in extramedullary relapsed/refractory B-acute lymphoblastic leukemia obtained with Inotuzumab after failure of treatment with Blinotumomab, thus showing differential target effects when using these antibodies.

The experimental section is opened by an article (Orlova et al.), concerning synthetic poly-L, L-lactide matrices which, being implanted with allogeneic mesenchymal stem cells, are able to promote restoration of the urinary bladder defects in the animals.

An article by Parfenenkova et al. deals with exosomes, the nanoparticles derived from tumor cells which, as shown in the in vitro experiments, may transfer oncogenic transcripts to the cultured normal mesenchymal cells from human hematopoietic microenvironment.

The CTT issue is closing with a review of actual studies reported at the XXIV Congress of European Hematology Association. The article covers new results and developments in hematopoietic stem cell transplantation, application of novel targeted drugs, including immune checkpoint inhibitors, in lymphoma treatment, CAR-T cell therapy an acute lymphoblastic leukemia, new approaches to treatment of myeloid neoplasias, as well as ensuing infectious problems and antibacterial treatment in these conditions.

Review articles

Clinical studies

Outcomes of allogeneic hematopoietic stem cell transplantation in children and adults with prior invasive fungal diseases

Marina O. Popova 1, Alisa G. Volkova 1, Inna V. Markova 1, Oksana V. Ayzsilnieks 1, Yulia A. Rogacheva 1, Anastasia S. Frolova 1, Aleksandr N. Shvetcov 1, Ilya Y. Nikolaev 1, Svetlana M. Ignatyeva 2, Tatyana S. Bogomolova 2, Asmic G. Gevorgian 1, Olesya V. Paina 1, Tatiana A. Bykova 1, Elena I. Darskaya 1, Maria V. Vladovskaya 1, Ivan S. Moiseev 1, Ludmila S. Zubarovskaya 1, Nikolay N. Klimko 1,2, Boris V. Afanasyev 1

Thrombopoietin receptor agonists for treatment of poor graft function after allogeneic hematopoietic stem cell transplantation in adults

Tatiana A. Rudakova, Alexander D. Kulagin, Ivan S. Moiseev, Tatyana A. Bykova, Sergey N. Bondarenko, Maria V. Barabanshikova, Anastasia V. Beinarovich, Anna A. Osipova, Varvara N. Ovechkina, Alexander L. Alyanskiy, Elena I. Darskaya, Elena V. Morozova, Boris V. Afanasyev

Clinical features and prognostic value of functional disorders and pulmonary comorbidity in adult patients prior to allogeneic hematopoietic stem cell transplantation

Egor A. Kulagin 1, Alisa G. Volkova 2, Ilya Yu. Nikolaev 2, Oleg V. Goloshchapov 2, Anna G. Smirnova 2, Tatiana A. Rudakova 2, Elena I. Darskaya 2, Elena V. Morozova 2, Nataliya B. Mikhaylova 2, Julia D. Rabik 3, Victoria G. Timchik 3, Tatiana I. Shchemelinina 3, Ruf D. Skvortsova 3, Tatiana S. Razumovskaya 3, Sergey N. Bondarenko 2, Ivan S. Moiseev 2, Valeriy N. Marchenko 1, Vasiliy I. Trofimov 1, Boris V. Afanasyev 2

Efficiency of mesenchymal stem cell therapy in ulcerative colitis as assessed by the morphology of colon mucosa

Galina V. Fedotovskikh, Galija M. Shaymardanova, Manarbek B. Askarov, Maiya S. Zhumabayeva, Gulmira S. Dosataeva, Aigerim K. Smagulova, Sapargul Marat, Tatyana G. Ezhelenko

Clinical significance of BAALC overexpression for predicting post-transplant relapses in acute myeloid leukemia

Alena I. Shakirova, Nikolay N. Mamaev, Ildar M. Barkhatov, Yana V. Gudozhnikova, Tatiana L. Gindina, Elena V. Babenko,
Boris V. Afanasyev

Clinical case

Successful treatment of extramedullary relapsed/refractory B-acute lymphoblastic leukemia with Inotuzumab ozogamicin before and after allogeneic stem cell transplantation

Anna G. Smirnova, Sergey N. Bondarenko, Ivan S. Moiseev, Inna V. Markova, Elena I. Darskaya, Irina K. Golubovskaya, Bella I. Ayubova, Elena V. Babenko, Ildar M. Barkhatov, Alexander L. Alyanskiy, Tatiana L. Gindina, Alexander D. Kulagin, Boris V. Afanasyev

Experimental studies

Experimental urinary bladder reconstruction using allogeneic tissue engineering products

Nadezhda V. Orlova1, Alexander N. Muraviov1,4, Tatjana I. Vinogradova1, Natalija M. Yudintceva2, Yulia A. Nashchekina2, Natalija V. Zabolotnih1, Alexander A. Lebedev1, Magomedsadik G. Sheykhov1, Piotr K. Yablonsky1,3

Exosomes as a promising tool for research and molecular diagnostics of myeloproliferative disorders

Anna N. Parfenenkova1, Ildar M. Barkhatov2, Anton A. Kremlev2, Olga A. Epifanovskaya2, Andrey N. Gorshkov3, Vera V. Vysochinskaya3, Mikhail P. Grudinin3, Anton A. Kornev4, Boris V. Afanasyev2

Congress reviews

Highlights of the 24th European Hematology Association Congress, Amsterdam 2019

Liudmila V. Fedorova, Kirill V. Lepik, Marina O. Popova, Yulia Yu. Vlasova, Olga V. Kudyasheva, Yulia A. Rogacheva, Elena E. Lepik, Valentina V. Porunova, Anna A. Osipova, Yury R. Zalyalov, Elena I. Darskaya, Inna V. Markova

Review articles

PD-1 receptor on immune cells, its expression and potential role in cancer therapy

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Dmitry S. Elezov 1, Igor V. Kudryavtsev 2,3,4

1 Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia
2 Department of Immunology, Institute of Experimental Medicine, St. Petersburg, Russia
3 Department of Oncoimmunology, N. N. Petrov National Medical Research Center of Oncology, St. Petersburg, Russia
4 Department of Fundamental Medicine, Far Eastern Federal University, Vladivostok, Russia

PD-1 is among key receptors conveying an inhibitory signal to T cells. Over last decade, PD-1 and its ligand PD-L1 draw much attention, due to high efficiency of therapy with PD-1/PD-L1 inhibitors in a number of malignant disorders. In this review article, we aimed to summarize current data on the PD-1 receptor expression in different immune cell subpopulations, like as its potential role in cellular antitumor response. Along with molecular structure and receptor-ligand interactions, the main attention is drawn to special features of PD-1 expression on the СD8+ T cell population which plays a key role in antitumor immune response. Some common changes of PD-1 expression levels during the cell activation and differentiation are considered, mainly, for the CD8+ T cells. Moreover, we discuss PD-1 expression on the surface of regulatory T cells, NK cells, invariant NKT cells, myeloid suppressor cells which may play an important role for anticancer immune response. When performing current therapy with PD-1/PD-L1 inhibitors, the mentioned populations may influence development of resistance to this mode of immune treatment. Therefore, a number of recent studies are directed for studying the PD-1/PD-L1 involvement into the immune regulation and to test prospects of their usage as biomarkers for clinical immune checkpoint therapy.

Keywords

PD-1, PD-L1, expression, T cells, antitumor therapy, checkpoint inhibitors.

Clinical studies

Outcomes of allogeneic hematopoietic stem cell transplantation in children and adults with prior invasive fungal diseases

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Marina O. Popova 1, Alisa G. Volkova 1, Inna V. Markova 1, Oksana V. Ayzsilnieks 1, Yulia A. Rogacheva 1, Anastasia S. Frolova 1, Aleksandr N. Shvetcov 1, Ilya Y. Nikolaev 1, Svetlana M. Ignatyeva 2, Tatyana S. Bogomolova 2, Asmic G. Gevorgian 1, Olesya V. Paina 1, Tatiana A. Bykova 1, Elena I. Darskaya 1, Maria V. Vladovskaya 1, Ivan S. Moiseev 1, Ludmila S. Zubarovskaya 1, Nikolay N. Klimko 1,2, Boris V. Afanasyev 1

1 Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Chair of Hematology, Transfusiology and Transplantology, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia
2 Dept. of Clinical Mycology, I. Mechnikov North-Western State Medical University, St. Petersburg, Russia

Invasive fungal diseases (IFD) are a major cause of morbidity and mortality in hematological patients, but prognosis of IFD has improved recently, due to introduction of new antifungals and efficient diagnostic procedures. Number of patients with IFD who are candidates for allogeneic hematopoietic stem cell transplant (allo-HSCT) increased. However, publications on the field are limited and there are no data on results of pediatric allo-HSCT patients with prior IFD. This study focuses on the outcomes of allo-HSCT in children and adults with prior IFD.

Patients and methods

In a prospective study, 504 allo-HSCT recipients were included from Jan 2013 to Jul 2016. The median age was 24 y.o. [2 months to 76 years] including 164 children (<18 yo) and 340 adults. The cohort included 52% male patients. Most patients (74.6%) were diagnosed with high-risk acute leukemia. Allo-HSCT from HLA-matched unrelated donors (MUD) was performed in 58.5%, from matched related donors (MRD), 22.5%; haploidentical HSCT was performed in 19% of patients, predominantly with RIC (67%). EORTC/MSG 2008 criteria for diagnosis and response to therapy were used. In the patients with pre-transplant lung lesions detected by CT scan, bronchoscopy with BAL was used. “Active IFD” means IFD diagnosed just before HSCT.

Results

Incidence of IFD before allo-HSCT was 15% (n=76). According to EORTS/MSG 2008 criteria, 90.8% of patients had probable and 9.2% presented with proven IFD. Etiology of IFD prior to HSCT was as follows: invasive aspergillosis (IA), 75%; invasive candidiasis (IC), 13%; mucormycosis (Mu), 4%; Pneumocystis pneumoniae (PCP), 1.3%, and combination of IA with Mu (2 cases), IC (n=1), PCP (n=1). The main sites of infection were lungs (95%), other localizations were predominantly combined with lung involvement: sinuses (9%), spleen (6%), liver (6%), and soft tissues (3%). Antifungal therapy before allo-HSCT was used in 75% patients with median duration of 2 months. Complete response to antifungal therapy was observed in 38.2% of the cases, partial response or stabilization was achieved in 35.5%, and “active IFD” was detected in 26.3% of the patients. After allo-HSCT, all the patients received antifungal therapy or secondary prophylaxis according to the IFD etiology. Cumulative incidence of relapse or progression of IFD after allo-HSCT was observed in 14.5%. Active underlying disease (hematology malignancy) at the moment of HSCT was the only risk factor for relapse or progression of IFD after allo-HSCT (11.5% vs 21.1%, p=0,03). We detected no significant differences in the cumulative incidence of acute, chronic GVHD and relapse in the study group as compared to the patients without history of IFD. 3-year overall survival (OS) after allo-HSCT was 67.5%. The impact of prior IFD on overall survival in allo-HSCT recipients was not statistically significant for the entire group (60.5% vs 68.7%, p=0.1), and, separately, for children (50.0% vs 57.4%, p=0,3) and adults (63.3% vs 74.6%, p=0.09). The worst outcome was observed in the patients with “active IFD” and active underlying disease at the moment of HSCT (3-year OS was 20%, p<0.001 compared to patients without history of IFD). However, in the patients with “active IFD” and remission of underlying disease, OS value was similar to survival rate of patients without history of IFD (80% vs 68.7%, p=0.2).

Conclusion

Incidence of IFD before allo-HSCT was 15%. Cumulative incidence of relapse or progression of IFD after allo-HSCT was 14.5%. Prior IFD had no significant impact on transplant-related complications and overall survival in children and adults undergoing allo-HSCT. Active underlying disease at the moment of HSCT was the only risk factor for relapse or progression of IFD and impaired outcome of allo-HSCT.

Keywords

Invasive fungal disease, pre-existing IFD, allogeneic hematopoietic stem cell transplantation, children and adults, clinical outcomes.

Clinical studies

Thrombopoietin receptor agonists for treatment of poor graft function after allogeneic hematopoietic stem cell transplantation in adults

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Tatiana A. Rudakova, Alexander D. Kulagin, Ivan S. Moiseev, Tatyana A. Bykova, Sergey N. Bondarenko, Maria V. Barabanshikova, Anastasia V. Beinarovich, Anna A. Osipova, Varvara N. Ovechkina, Alexander L. Alyanskiy, Elena I. Darskaya, Elena V. Morozova, Boris V. Afanasyev

Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Chair of Hematology, Transfusiology and Transplantology, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia

Posttransplant cytopenias due to poor graft function (PGF) are a severe complication of allogeneic transplantation of hematopoietic stem cells (allo-HSCT). Conventional therapeutic options for poor graft function are limited to infusions of CD34-selected cells. However, having shown encouraging results in idiopathic thrombocytopenia and aplastic anemia, thrombopoietin receptor agonists (TPO-RA) romiplostim and eltrombopag might be a possible alternative treatment strategy for PGF after allo-HSCT. Current study aims to summarize single-center experience of TPO-RA for treatment of posttransplant cytopenias including PGF after allo-HSCT.

A total of 31 patients (15 males, 16 females) with median age of 22 years (18-57) received therapy with TPO agonists for posttransplant thrombocytopenia <20×109/l (n=7) and PGF (n=24). Romiplostim (n=17) with the median dose 5 mkg/kg weekly (3-5) or eltrombopag (n=14) with the median dose 50 mg/day (50-150) were used. Criteria for severe poor graft function (sPGF) were the following: cytopenia in two or more myeloid lineages (platelets <20×109/l, absolute neutrophil count (ANC) <0.5×109/l, hemoglobin <70 g/l) any time after documented engraftment, full or stable mixed donor chimerism >90% without signs of relapse of underlying disease. Overall response included complete response (CR) (platelets ≥100×109/l, ANC ≥1.5×109/l, hemoglobin ≥100 g/l) and partial response (PR) (platelets >20×109/l, ANC ≥0.5×109/l, hemoglobin >70 g/l). Median time from diagnosis of sPGF to the start of TPO-RA therapy was 14 days (0-119), median length of TPO-RA therapy was 3 weeks (1-43). TPO-RA was well tolerated, with no signs of II-IV grade toxicity. In a total of 8 cases (28%) TPO-RA were used in combination with rituximab (n=4), rituximab and donor lymphocyte infusions (DLI) (n=3), as well as CD34-cell infusions (n=1). A total of 14 (48%) patients achieved response (CR: n=4, 14%; PR: n=10, 34%). Combination therapy showed no advantage in response rate compared to TPO-RA alone. Median increase of ANC in responders was 3.4×109/l (0.8-6.0), platelets – 48×109/l (21-205). A total of 15 patients (3 responders, 12 non-responders) died of infection (n=10), relapse (n=2) and acute graft-versus-host disease (aGVHD) III-IV grade (n=3). One-year overall survival (OS) from the therapy start was 60% (95% CI, 40-76), with a significant difference between responders and non-responders: 79% (95% CI, 47-93) vs 39% (95% CI, 14-63) (p=0.01).

Our study showed a promising efficacy and safety of TPO-RA in patients with sPGF after allo-HSCT. Multicenter prospective studies are needed to establish optimal dose and duration of the drug administration, as well as the criteria for discontinuation of these drugs.

Keywords

Allo-HSCT, poor graft function, thrombopoietin receptor agonists.

Clinical studies

Clinical features and prognostic value of functional disorders and pulmonary comorbidity in adult patients prior to allogeneic hematopoietic stem cell transplantation

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Egor A. Kulagin 1, Alisa G. Volkova 2, Ilya Yu. Nikolaev 2, Oleg V. Goloshchapov 2, Anna G. Smirnova 2, Tatiana A. Rudakova 2, Elena I. Darskaya 2, Elena V. Morozova 2, Nataliya B. Mikhaylova 2, Julia D. Rabik 3, Victoria G. Timchik 3, Tatiana I. Shchemelinina 3, Ruf D. Skvortsova 3, Tatiana S. Razumovskaya 3, Sergey N. Bondarenko 2, Ivan S. Moiseev 2, Valeriy N. Marchenko 1, Vasiliy I. Trofimov 1, Boris V. Afanasyev 2

1 M. Chernorutskiy Department of Hospital Therapy, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia
2 Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia
3 Department of Functional Diagnostics №2, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia

High efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in a wide range of malignant and non-malignant hematopoietic disorders system is limited to the risk of serious complications and non-relapse mortality (NRM). Pre-existing comorbidities before allo-HSCT are among key factors for NRM.

The purpose of this study was to characterize the potential pre-transplant factors of pulmonary comorbidity and integral indicators of pulmonary function tests (PFT) and to evaluate their prognostic value for the allo-HSCT outcomes in adult patients.

Patients and methods

The study included 355 patients, among them 149 (42%) in the prospective phase of the study. The cohort included patients with acute leukemia (60%), myeloproliferative diseases (10%), myelodysplastic syndrome (5%), lymphomas (17%), and severe aplastic anemia (8%). The median age at the time of allo-HSCT was 33 years old (18 to 66). We analyzed all the factors that could affect lung function before allo-HSCT. The severity of pulmonary comorbidity was assessed on the basis of forced expiratory volume in one second (FEV1) and diffusing capacity of carbon monoxide (CO) (DLco), in accordance with the criteria of the Hematopoietic Cell Transplantation Specific Comorbidity Index (HCT-CI). The prognostic value of pulmonary comorbidity was investigated with respect to the cumulative incidence of mechanical ventilation (MV) and NRM rates.

Results

Tobacco smoking (32%), history of chronic obstructive pulmonary disease (COPD) and bronchial asthma (BA) (5%), previous pulmonary infections (45%), potentially pulmonary toxic chemotherapy (79%), previous auto-HSCT (8%), usage of immune checkpoint inhibitors (5%), mediastinal radiation therapy (6%) were the most common pulmonary compromising factors before allo-HSCT. According to the HCT-CI criteria, 90 patients (25%) had low FEV1 values, with 14%, 9% and 2% of mild, moderate and severe grade respectively. DLco disorders were diagnosed in 69% of the patients examined, including 29%, 28%, and 12% of mild, moderate, and severe grade respectively. Cumulative incidence of MV within 100 days after allo-HSCT was 7.2%, 10.9%, 16.7%, and 22.1% in patients with pulmonary comorbidity of 0, 1, 2, 3 degrees, respectively. The degree of pulmonary comorbidity was an independent risk factor for NRM (hazard ratio 1.39; CI 95%; 1.03-1.89; p=0.033), being associated with a decrease in overall survival (OS).

Conclusions

Adult patients have a wide range of pulmonary comorbidity factors before allo-HSCT. HCT-CI pulmonary comorbidity is an independent risk factor for MV during the first 100 days after allo-HSCT and NRM.

Keywords

Allogeneic hematopoietic stem cell transplantation, adults, pulmonary comorbidity, pulmonary functional tests, clinical prognosis.

Clinical studies

Efficiency of mesenchymal stem cell therapy in ulcerative colitis as assessed by the morphology of colon mucosa

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Galina V. Fedotovskikh, Galija M. Shaymardanova, Manarbek B. Askarov, Maiya S. Zhumabayeva, Gulmira S. Dosataeva, Aigerim K. Smagulova, Sapargul Marat, Tatyana G. Ezhelenko

National Scientific Medical Center, Nur-Sultan (Astana), Kazakhstan

Usage of bone marrow mesenchymal stromal cells (MSCs) in human inflammatory bowel disease has proven its feasibility in clinical studies. The purpose of this work was to study morpho-functional state of large intestine mucosa (LIM) at the term of 10 months after a single infusion of cultured in vitro expanded mesenchymal bone marrow cells (MSC) added to standard therapy of patients with ulcerative colitis (UC).

Materials and methods

Initial group of the patients with UC included 18 subjects. Of those, nine patients received standard drug therapy (control group), six subjects underwent similar therapy supplied by a single transplantation of autologous MSCs (the study group). Clinical and functional state of the patients was assessed by indices, according to Truelove and Wiits, Mayo and Rakhmilevich. Routine light microscopy, as well as electron microscopic studies were performed for 320 large intestinal mucosa (LIM) biopsies. Specific histological and immunohistochemical examination was carried out in 64 samples. Histological signs of inflammatory activity were evaluated in points according to K.Geboes et al. (2000). Immunohistochemistry studies included expression of CD4, CD68, CD31, CD34, CD8, lgG, and Ki-67 markers. The electron microscopic technique was used as generally accepted. Semi-thin sections were stained by C. Humphrey and F. Pittman (1974). Ultrathin sections were observed and captured using a Libra 120 electron microscope.

Results

A single transplantation of autologous cultured bone marrow MSCs, in addition to standard therapy of patients with ulcerative colitis has generally improved clinical and laboratory signs of the disease as examined 10 months after treatment, when compared with initial clinical state and the control group. Upon morphological study of LIM in the study and control groups, erosive and ulcerative defects did persist or were slightly decreased. Indicators of positive dynamics in the LIM structure have manifested mainly in the patients of study group, as a decreased inflammatory response and restoration of intestinal crypts (“epithelial niche”), with increased number of goblet cells and decreased permeability of intestinal cover epithelium.

Conclusion

Single transplantation of autologous cultured bone marrow MSC could be implemented into the standard therapy of patients with ulcerative colitis, providing a positive effect upon morpho-functional state of LIM, thus promoting reduction of inflammatory response, recovery of crypts and cover epithelium.

Keywords

Ulcerative colitis, autologous bone marrow, mesenchymal cells, transplantation, intestinal crypts, morphology.

Clinical studies

Clinical significance of BAALC overexpression for predicting post-transplant relapses in acute myeloid leukemia

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Alena I. Shakirova, Nikolay N. Mamaev, Ildar M. Barkhatov, Yana V. Gudozhnikova, Tatiana L. Gindina, Elena V. Babenko,
Boris V. Afanasyev

Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia

The aim of this study was to clarify close associations between population of bone marrow blast elements defining cytological relapse, and that of leukemic precursor cells by means of serial quantitative measurement of WT1 and BAALC molecular markers expressed by appropriate cell types. In clinical terms, we sought to evaluate a potential for evolving leukemia relapses following hematopoietic stem cell transplantation (HSCT). The post-transplant relapses (PTR), in view of their biological mechanisms, may be detected by overexpression of distinct differentiation-related genes.

Patients and methods

We have done parallel serial measurements of BAALC and WT1 gene expression in bone marrow cells of the patients with acute myeloid leukemia, using quantitative real-time PCR (qPCR) with gene-specific primers.

Results

An initial pilot study was carried out in twelve patients with elevated WT1 expression levels and normal blast counts (<5%) in bone marrow at the time of allogeneic HSCT. It has shown that the relapse occurrence and shorter relapse-free survival (RFS) post-transplant were more common in the subgroup with higher BAALC gene expression level as compared to those without BAALC expression elevation (p=0.002, and p=0.019, respectively). Moreover, when analyzing the validating patient cohort, we have found that the simultaneous elevation of both BAALC and WT1 expression, known as unfavorable prognostic factor, was more frequently observed in the patients with AML М1, М2, and М0 FAB-variants. Preliminary results show an inferior overall survival (a mean of 228 days) in AML patients with combined overexpression of the both genes immediately before HSCT, like as in post-transplant period. In conclusion, it should be noted that our findings of simultaneous overexpressions of BAALC and WT1 genes in great cohort of adults and children patients with different FAB- variants of AML treated by means of allo-HSCT were revealed for the first time. Being associated with poor prognosis, this double test should be used actively for post-transplant monitoring of all patients with AML.

Keywords

Acute myeloblastic leukemia, hematopoietic stem cell transplantation, allogeneic, post-transplant relapses, BAALC, WT1, gene overexpression, parallel assays, relapse risk, prognostic value.

Clinical case

Successful treatment of extramedullary relapsed/refractory B-acute lymphoblastic leukemia with Inotuzumab ozogamicin before and after allogeneic stem cell transplantation

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Anna G. Smirnova, Sergey N. Bondarenko, Ivan S. Moiseev, Inna V. Markova, Elena I. Darskaya, Irina K. Golubovskaya, Bella I. Ayubova, Elena V. Babenko, Ildar M. Barkhatov, Alexander L. Alyanskiy, Tatiana L. Gindina, Alexander D. Kulagin, Boris V. Afanasyev

Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Chair of Hematology, Transfusiology and Transplantology, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia

Recent advances in treatment of relapsed/refractory B-Acute Lymphoblastic Leukemia (R/R B-ALL) are associated with targeted immune therapies, such as Blinatumomab, Inotuzumab ozogamicin and CAR-T cells. Several clinical studies demonstrate superior complete remission (CR) and allogeneic stem cell transplantation (allo-HSCT) rates after Blinatumomab or Inotuzumab treatment compared to conventional chemotherapy with different toxicity profiles. However, treatment of extramedullar lesions was not addressed in these trials.

Case presentation

We report two R/R B-ALL patients with extramedullar involvement. Case 1. A woman, 31 years old with Philadelphia positive B-ALL, being in second relapse not responding to Blinotumomab. Clinical progression was observed, with involvement of bone marrow (BM), cervical, mandibular lymph nodes and breast lesions. Complete BM and extramedullary remission was achieved after Inotuzumab followed by allogeneic stem cell transplantation from a matched related donor was performed. Case 2. A 25 years-old young woman exhibited a BM relapse after unrelated stem cell transplantation. Remission was achieved after Blinotumomab treatment. Inotuzumab was administered for 6th extramedullary relapse. A complete metabolic response was achieved after 4 cycles of Inotuzumab as shown by PET scanning.

Conclusion

The present cases demonstrate an opportunity for Inotuzumab administration after Blinotumomab failure in case of extramedullary relapse before and after allo-HSCT.

Keywords

Allogeneic hematopoietic stem cell transplantation, B-Acute Lymphoblastic Leukemia, Blinotumomab, Inotuzumab ozogamicin, extramedullary relapse.

Experimental studies

Experimental urinary bladder reconstruction using allogeneic tissue engineering products

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Nadezhda V. Orlova1, Alexander N. Muraviov1,4, Tatjana I. Vinogradova1, Natalija M. Yudintceva2, Yulia A. Nashchekina2, Natalija V. Zabolotnih1, Alexander A. Lebedev1, Magomedsadik G. Sheykhov1, Piotr K. Yablonsky1,3

1 St. Petersburg State Research Institute of Phthisiopulmonology, St. Petersburg, Russia
2 Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia
3 St. Petersburg State University, St. Petersburg, Russia
4 Private University «St. Petersburg Medico-Social Institute», St. Petersburg, Russia

The article describes the results of an experimental study of tissue-engineering constructions based on lactic acid – poly-L, L-lactide matrix, containing allogeneic cells of various tissue origin.

Aim

Experimental investigation of the possibility of using an allogenic tissue-engineering graft to replace a defect in the bladder wall.

Materials and methods

The study was performed on 15 male Chinchilla rabbits. After partial resection of the bladder augmentation cystoplasty was performed with scaffolds containing smooth myocytes with urothelium, fibroblasts and mesenchymal stem cells.

Results

In 100% of cases of partial bladder replacement with a cell-free matrix or scaffolds containing smooth myocytes with urothelium and fibroblasts, an implant was rejected with a different severity of the inflammatory reaction and a decrease in the capacity of the bladder. In the 4th group (mesenchymal stem cells), on the contrary, in 5 cases out of 6 lysis of the matrix occurred, the capacity of the bladders 2.5 months post-surgery was comparable to the preoperative one. At the implantation site, an area of the modified mucous membrane with signs of vascularization was determined. Histologically, the initial stages of reparation and angiogenesis were revealed. With confocal microscopy of cryosections at the implantation site, labeled cells are identified that are involved in the formation of a structure similar to the urothelium.

Conclusion

The efficiency of using mesenchymal stem cells as part of a tissue-engineered product for partial replacement of the bladder wall was shown. Development of methods of creating multi-component graft using allogeneic cells may improve the results of treatment of pathologies in which no autologous material is available.

Keywords

Allogeneic transplantation, bone marrow mesenchymal stromal cells, bladder, tissue engineering.

Experimental studies

Exosomes as a promising tool for research and molecular diagnostics of myeloproliferative disorders

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Anna N. Parfenenkova1, Ildar M. Barkhatov2, Anton A. Kremlev2, Olga A. Epifanovskaya2, Andrey N. Gorshkov3, Vera V. Vysochinskaya3, Mikhail P. Grudinin3, Anton A. Kornev4, Boris V. Afanasyev2

1 St. Petersburg State University of Chemical Pharmaceutics, St. Petersburg, Russia
2 Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia
3 A. Smorodintsev Research Institute of Influenza, St. Petersburg, Russia
4 St. Petersburg National Research University, Russian Academy of Sciences, St. Petersburg, Russia

Detection of molecular markers characteristic to malignant cells is the key task of molecular genetic studies in oncohematology, aiming for timely diagnosis of the diseases and evaluation of their progression, like as relapse risks. Exosomes are among the most promising objects for such studies. They represent a fraction of extracellular membrane particles ranging from 30 to 150 nm in size, presumably containing such tumor markers. These vesicles are found in blood serum and other biological fluids, being important participants in cell-cell interactions capable of transferring various biopolymers, which can also mediate the reprogramming of the functions of the recipient cells and influence the development of resistance to therapy. On the basis of exosomes, one may optimize such a novel diagnostic approach as fluid biopsy which is minimally invasive, and, therefore, can also be used as a specific monitoring tool for prediction of tumor relapse development.

As a part of this work, we aimed for finding whether exosomes can potentially influence the results of minimal residual disease determination in the patients with chronic myeloid leukemia (CML).

In order to study the characteristics of exosomes, we used the method of sequential ultracentrifugation followed by analysis of the morphological and functional characteristics of exosomes. In addition, the possibility of horizontal transfer of the chimeric BCR-ABL transcript from leukemic to stromal cells through the exosomal fraction was analyzed.

The results of our in vitro study show evidence of horizontal transfer of oncogene mRNA from malignant to normal stromal cells. In conclusion, considering the fact that bone marrow mesenchymal stromal cells (MSCs) play the main role in hematopoiesis regulation, which forms a niche for hematopoietic stem cells (HSCs), we suggested that extracellular vesicles may act as a possible source of detectable oncogene transcripts and their migration into the cells of microenvironment.

Keywords

Extracellular vesicles, exosomes, gene overexpression, molecular markers, cell-cell interactions, mesenchymal stem cells, mRNA transport, BCR-ABL, chronic myeloid leukemia, minimal residual disease, liquid biopsy.

Congress reviews

Highlights of the 24th European Hematology Association Congress, Amsterdam 2019

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Liudmila V. Fedorova, Kirill V. Lepik, Marina O. Popova, Yulia Yu. Vlasova, Olga V. Kudyasheva, Yulia A. Rogacheva, Elena E. Lepik, Valentina V. Porunova, Anna A. Osipova, Yury R. Zalyalov, Elena I. Darskaya, Inna V. Markova

Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov First Saint Petersburg State Medical University, St. Petersburg, Russia

24th Congress of European Hematology Association was held from 13th to 16th July in Amsterdam, Netherlands. Over 12000 specialists took part in the meeting and more than 360 oral and 1300 poster presentations were made, covering most of hematology fields. The main purpose of this article was to review the most important and relevant topics discussed during congress in such areas as hematopoietic stem cell transplantation, lymphomas, acute leukemia, multiple myeloma and other plasma cell disorders, myeloproliferative neoplasms and infectious complications in hematological patients.

In this review, we briefly describe results of works which, in view of the authors, are of most clinical and scientific significance for hematopoietic stem cell transplantation. Moreover, the emerging topics and trends in different fields of hematology were discussed. Other interesting studies and presentations can be accessed through the EHA online-library.

Keywords

EHA congress, hematopoietic stem cell transplantation, lymphoma, acute leukemia, multiple myeloma, myeloproliferative neoplasms, infectious complications.