ISSN 1866-8836
Клеточная терапия и трансплантация

Volume 9, number 2
07/30/2020
Volume 9, number 2
Editor-in-Chief
Kulagin A. D. (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)
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

Editorial article

Editorial article

Professor Alexander D. Kulagin, Editor-in-Chief, Cellular Therapy and Transplantation Journal

Review articles

CaCO3 vaterites as components of target drug delivery systems

Natalia N. Sudareva1,2, Pavel V. Popryadukhin1,2, Natalia N. Saprykina1, Olga M. Suvorova1, Galina Yu. Yukina2, Oleg V. Galibin2, Aleksandr D. Vilesov1,2

Clinical studies

Clinical efficiency and safety of tramadol and low-dose morphine to manage pain syndromes in children following chemotherapy and hematopoietic stem cell transplantation

Ekaterina V. Goncharova1,2, Inga E. Zavodova1, Nikita P. Volkov1, Olga A. Ivanova1, Maxim A. Kucher1, Alexey Y. Sokolov2,3, Maxim P. Bogomolny1, Gleb E. Ulrikh4, Ludmila S. Zubarovskaya1, Boris V. Afanasyev1

Ratio of total leukocyte count to C-reactive protein: does it help to differentiate infectious fever from engraftment fever in patients undergoing autologous hematopoietic stem cell transplant?

Sachin Punatar1,2, Lingaraj Nayak1,2, Avinash Bonda1,2, Anant Gokarn1,2, Aniket Mohite1, Karthik Shanmugam1, Deepan Rajamanickam1, Alok Gupta1, Libin Mathew1, Sadhana Kannan3, Navin Khattry1,2

Bacteroides fragilis is a potential marker of effective microbiota transplantation in acute graft-versus-host disease treatment

Oleg V. Goloshchapov1, Evgenyi A. Bakin1, Maxim A. Kucher1, Oksana V. Stanevich1, Maria A. Suvorova2, Vladimir V. Gostev3, Oleg S. Glotov4, Yury A. Eismont4, Dmitry E. Polev5, Anastasia Yu. Lobenskaya5, Ruslana V. Klementeva1, Maria O. Goloshchapova1, Ludmila S. Zubarovskaya1, Sergey V. Sidorenko3, Alexander N. Suvorov6, Ivan S. Moiseev1, Alexei B. Chukhlovin1

Morphological skin evaluation in the patients with systemic scleroderma before and after hematopoietic stem cell transplantation

Galina V. Fedotovskikh, Galija M. Shaymardanova, Manarbek B. Askarov, Ainash A. Zhusupova, Natalya A. Krivoruchko, Tatyana G. Ezhelenko, Sapargul Marat

Clinical case

A long-term response to allogeneic hemopoietic stem cell transplantation from haploidentical donor and post-transplant therapy in an adolescent with primary resistant neuroblastoma

Ilya V. Kazantsev1, Tatiana V. Iukhta1, Asmik G. Gevorgian1, Polina S. Tolkunova1, Andrew V. Shamin2, Vadim V. Baykov3, Nikolay A. Vorobyov4, Andrew V. Kozlov1, Marina A. Karsakova2, Polina S. Kuga1, Alexander N. Shvetsov1, Elena V. Morozova1, Svetlana S. Safonova1, Yuri A. Punanov1, Ludmila S. Zubarovskaya1, Boris V. Afanasyev1

Successful treatment of relapsed/refractory B-Acute lymphoblastic leukemia with Inotuzumab ozogamicin after blinatumomab failure

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

Experimental studies

Augmented reality technology for auricular reconstruction in the treatment of microtia

Аndrey I. Yaremenko1, Anna V. Lysenko1, Elizaveta A. Ivanova1, Oleg V. Galibin2

Obituary

In memory of Professor Rolf Neth October 6, 1926 – March 17, 2020

Boris Fehse, Nicolaus Kröger, Carol Stocking, Axel Zander

Rolf Neth and Russia

Prof. Margarita B. Belogurova,
Department of Pediatric Oncology, City Hospital No. 31, St. Petersburg, Russia

Prof. Ludmila S. Zubarovskaya,
RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia

Editorial article

Editorial article

Download PDF version

Professor Alexander D. Kulagin, Editor-in-Chief, Cellular Therapy and Transplantation Journal

Dear CTT authors and readers,

The initial design for this issue of Cellular Therapy and Transplantation (CTT) was prepared by the Editor-in-Chief, Professor Boris V. Afanasyev who untimely passed away on March 16, 2020. The CTT edition was one of his priorities, which made the journal a recognizable tool of international cooperation, a forum for discussing the most difficult clinical issues and challenging aspects of oncology, hematology, stem cell transplantation, and relevant areas of immunology, molecular biology, cellular and gene therapy.

A wide range of problems highlighted in CTT since 2008, was determined by integrative role of hematopoietic stem cell transplantation (HSCT) which applies multidisciplinary diagnostic approaches to improve safety and efficiency of the procedure. This progress in HSCT is based on huge number of new fundamental data obtained in adjacent fields of biology and medicine, then translated into clinical achievements. Now we are more informed about the mechanisms underlying the development of all severe clinical situations in which hematopoietic cell transplants are performed, including leukemias, lymphomas, other conditions with clonal hematopoiesis, bone marrow failure syndromes, immune deficiencies, autoimmune disorders, hereditary diseases. Molecular biology studies enabled us to detect marker genes which could be used for improved diagnostics and risk stratification of the patients.

New options of targeted pharmacological control and immunotherapy permitted us more accurately determine indications for HSCT, thus increasing survival and minimizing possible adverse effects from the treatment. Therefore, the scientific priorities and topics of CTT journal represent a response to the growing needs for new knowledge, wide and effective interdisciplinary interaction.

An essential feature and advantage of the journal is the coverage of new clinical data in both pediatric and adult patients. A constant exchange of knowledge and comparative experience gained in seek children and adults, is mutually enriching and absolutely necessary when assessing the role of age factor in clinical oncology, hematology and hematopoietic cell transplantation. This is another notable tradition which stems from the history of founding and development of Raisa Gorbacheva Memorial Research Institute of Pediatric Oncology, Hematology and Transplantation established at the Pavlov University in 2007.

The journal is a permanently developing edition, and it presents the whole range of scientific publications – scientific reviews, original clinical and experimental studies, guidelines, and descriptions of rare clinical observations. Among CTT authors, you will always find widely known experts covering major theoretical and clinical problems, along with young researchers and clinicians who focus on their individual topics. This principle is one of the key CTT traditions established by Professor Boris V. Afanasyev, who attached great importance to continuity of generations in fundamental research and clinical practice.

CTT journal traditionally highlights and publishes abstracts of the Raisa Gorbacheva Memorial Meeting Hematopoietic Stem Cell Transplantation. Gene and Cellular Therapy. Over 14 years, the Symposium has become a renowned scientific forum in the field of blood cancer treatment and hematopoietic cell transplantation. This year, our symposium will take place with the active use of online format on a specially developed platform with virtual and real participation of leading Russian and foreign experts in the field of hematopoietic stem cell transplantation and cellular therapy. Despite a required social distancing and difficulties of the face-to-face participation, we again expect a large audience and interesting scientific discussions.

During these hard times for the medical community, the editors wish good health to all our readers and authors, and, especially, faster overcoming the pandemic, hoping for opportunities of future scientific meetings, as well as discussing new experimental data and current clinical problems of hematology, transplantation and adjacent research fields on the pages of Cellular Therapy and Transplantation.

Review articles

Can immune therapy cure acute myeloid leukemia?

Download PDF version

Robert P. Gale

Imperial College London, London, UK


Correspondence
Robert Peter Gale MD, PhD, DSc(hc), FACP, FRCP, Visiting Professor Haematology, Centre for Haematology Research, Department of Immunology and Inflammation Imperial College London, London, UK
E-mail: robertpetergale@gmail.com

There is considerable progress in immune therapy of diverse cancers. In haematology these advances are mostly limited to lymphoid cancers. Effective therapies include monoclonal antibodies and chimeric antigen receptor (CAR)-T-cells to lymphoid lineage-antigens such as CD19, CD20 and B-cell maturation antigen (BCMA). Gemtuzumab ozogamicin (Myelotarg®) is the only FDA-approved immune-based therapy for acute myeloid leukemia (AML). Several clinical trials of antibodies to CD38 and CD123 are reported with unimpressive efficacy and safety concerns. Reasons are higher daily production rates of myeloid cells and unacceptable collateral damage to normal haematopoietic cells because of imperfect specificity for AML cells. Potential targets of anti-AML immune therapy are (1) HLA antigens; (2) minor histocompatibility antigens; (3) leukemia-associated antigens; and (4) leukemia-specific antigens. Data supporting an effective allogeneic anti-AML effect come from studies in recipients of haematopoietic cell transplants with graft-versus-host disease (GvHD) and recipients of donor lymphocyte infusions (DLI). A special problem is a relative paucity of neo-antigens in AML compared with solid cancers because of a low cumulative mutation frequency. Cell immune therapy trials are in progress including CAR-T-cells, CAR-NK-cells and allogeneic NK-cells. Approaches using synthetic biology are being developed. Presently, except for gemtuzumab ozogamicin there are no convincing data of efficacy of immune therapy in AML.

Keywords

Acute myeloid leukemia, mutations, neoantigens, immune therapy.

Review articles

CaCO3 vaterites as components of target drug delivery systems

Download PDF version

Natalia N. Sudareva1,2, Pavel V. Popryadukhin1,2, Natalia N. Saprykina1, Olga M. Suvorova1, Galina Yu. Yukina2, Oleg V. Galibin2, Aleksandr D. Vilesov1,2

1 Institute of Macromolecular Compounds RAS, St. Petersburg, Russia
2 Pavlov University, St. Petersburg, Russia


Correspondence
Dr. Natalia N. Sudareva, Research Institute of Macromolecular Compounds, Bolshoi Prosp. 31 (V.O.), 199004, St. Petersburg, Russia
E-mail: nnsas@mail.ru

Successful treatment of the majority of oncological diseases that affect solid organs is related to appropriate use of potent and (to varying degrees) toxic antitumor drugs. In a number of cases, chemotherapy requires the maximum localized action of a drug in the tumor area. The most efficient methods of drug administration are introducing medicinal compounds (MC) directly into the tumor or use of target drug delivery systems. The second method makes it possible to decrease general toxicity of MC, and to reach prolonged therapeutic action due to uniform and time-controlled release of a MC into tumor tissue.

In the present work, we studied behavior of porous spherical СаСО3 vaterites (components of delivery systems for antitumor drugs) in various environments (human blood plasma, rat muscle tissue). It was demonstrated that the studied drug carriers undergo morphological transformations and are destructed with time. In blood plasma, due to ion exchange reactions, vaterites are transformed into gradually disintegrating needle-like structures (as shown by scanning electron microscopy and energy dispersive spectroscopy). Similar processes were observed in muscle tissue: in three days, spheres were transformed into needle-like structures and then underwent complete bioresorption.

Keywords

Anticancer drugs delivery systems, СаСО3 vaterites, blood plasma, intramuscular administration, bioresorption.

Clinical studies

Attitudes to the disease and therapy in patients with chronic Ph-negative myeloproliferative neoplasms: results of the physician and patient surveys in Russia as a part of International Landmark Study

Download PDF version

Elena V. Morozova1, Maria V. Barabanshchikova1, Tatyana I. Ionova2, Boris V. Afanasyev1

1 RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia
2 Saint Petersburg State University Hospital, St. Petersburg, Russia


Correspondence
Prof. Dr Sci Tatyana I. Ionova, Saint Petersburg State University Hospital, Fontanka Emb 154, 198103, St. Petersburg, Russia
Phone: +7 (962) 710 1711
E-mail: tation16@gmail.com

The aim of this paper was to present evaluation and synthesis of data derived from a survey of Russian patients and physicians, performed as a part of the international Landmark study for the emerging market countries designed to specify problems and areas of concern in management of patients with chronic Ph-negative myeloproliferative neoplasms (MPN). The online survey forms were filled by 40 adult patients with Ph(-) MPNs (PV, 42.5%; MF, 37.5%; ET, 20%) and 30 physicians with sufficient experience in the Ph(-) MPNs treatment. As a part of this survey, patients and physicians answered questions related to perception of the disease symptoms and their impact on quality of life, daily activities and work productivity of patients, as well as their attitude to main treatment goals and various aspects of the patient-physician communication. The results revealed a number of differences between patient and physician perception of the disease and treatment, thus complementing the data of the Landmark Survey in other countries. It was shown that the patients with different variants of Ph(-) MPNs encounter sufficient disease-related difficulties in everyday life, impaired quality of life and reduced work productivity. Lack of coincidence revealed between the physician and patient assessment of the disease burden and treatment indicates the need for new ways of improving quality of clinical care provided to this category of patients. Further research in this area would be an important step towards implementation of patient-centered Ph(-) MPN treatment programs in Russian Federation.

Keywords

Myeloproliferative neoplasms, chronic, Ph-negative, physician and patient survey, symptoms, quality of life, patient-centered treatment programs.

Clinical studies

Clinical efficiency and safety of tramadol and low-dose morphine to manage pain syndromes in children following chemotherapy and hematopoietic stem cell transplantation

Download PDF version

Ekaterina V. Goncharova1,2, Inga E. Zavodova1, Nikita P. Volkov1, Olga A. Ivanova1, Maxim A. Kucher1, Alexey Y. Sokolov2,3, Maxim P. Bogomolny1, Gleb E. Ulrikh4, Ludmila S. Zubarovskaya1, Boris V. Afanasyev1

1 RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia
2 Department of Neuropharmacology, Valdman Institute of Pharmacology, Pavlov University, St. Petersburg, Russia
3 Pavlov Institute of Physiology of the Russian Academy of Sciences, St. Petersburg, Russia
4 Department of Anesthesiology and Pediatric Intensive Care, Saint Petersburg State Pediatric Medical University, St. Petersburg, Russia


Correspondence
Dr. Ekaterina V. Goncharova, RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, L.Tolstoy St. 6-8, 197022, St. Petersburg, Russia
Phone: +7 (911) 087 8976
E-mail: ek.v.goncharova@gmail.com

A sufficient subgroup of patients encounters pain syndrome in the course of cytostatic chemotherapy (ChT), either with or without hematopoietic stem cell transplantation (HSCT). Over this time period, severe thrombocytopenia and leucopenia may develop, thus limiting the opportunities for non-steroidal anti-inflammatory drugs (NSAID). As recommended by WHO, administration of strong opioids to children is possible in moderate pain and inefficiency of NSAIDs. In this case, second step of the pain relief ladder is absent, i.e., codeine application. However, the recommendations do not exclude usage of tramadol, which is widely applied in pediatrics. Our aim was to evaluate relative safety and efficiency of tramadol and morphine in managment of moderate pain in children after HSCT and ChT.

Patients and methods

The study included analysis of 159 children admitted to the ICU pain management team with complaints for weak or moderate pain (form 3 to 6 points on an age-matched scale). The age of patients was from 1 to 17 years, with a median of 8 years old. All the patients did not receive opioids (were opioid naïve) within 30 days before inclusion to the study. The drugs were injected by continuous infusion at the inpatient clinic. In the first group (n=118), standard tramadol doses were administered as the 1st-line therapy (0.2 to 0.3 mg/kg/h). The patients form 2nd group (n=41) were administered low-dose morphine (0.01 to 0.019 mg/kg/h). Treatment efficiency was assessed by FLACC verbal scores, Wong-Baker Faces Pain Rating Scale, or visual analogue scale and quality of life. Statistical evaluation was performed by means of SPSS software, using a nonparametric Chi-square criterion.

Results

When administered tramadol as a first-line therapy, it was efficient in ca. 40.7% of cases (n=48). With low-dose morphine, the response rate proved to be 58.5% (n=24). One patient (0.8%) received tramadol when transferred to other institution. The second-line therapy (strong opioids) was administered due to lack of efficiency, or poor drug acceptability during the first-line treatment. It was observed in 53.4% of group 1 (n=63), and in 39% (n=16) of morphine-treated patients (group 2). Side effects due to tramadol administration were observed in 5.1% of cases (n=6). When administered low-dose morphine, only 1 female patient (2.4%) developed intestinal paresis which resolved after the therapy cancellation. Upon statistical evaluation, no significant differences were revealed between the groups.

Conclusion

Both medical drugs have shown similar efficiency and safety when applied for jugulating weak or moderate nociceptive pain after cytostatic chemotherapy and HSCT in pediatric patients.

Keywords

Chemotherapy, anticancer, pain syndrome, mucositis, tramadol, morphine, efficiency, safety.

Clinical studies

Ratio of total leukocyte count to C-reactive protein: does it help to differentiate infectious fever from engraftment fever in patients undergoing autologous hematopoietic stem cell transplant?

Download PDF version

Sachin Punatar1,2, Lingaraj Nayak1,2, Avinash Bonda1,2, Anant Gokarn1,2, Aniket Mohite1, Karthik Shanmugam1, Deepan Rajamanickam1, Alok Gupta1, Libin Mathew1, Sadhana Kannan3, Navin Khattry1,2

1 HSCT unit, Department of Medical Oncology Tata Memorial Centre, HSCT unit, ACTREC, Kharghar, Navi Mumbai, India
2 Homi Bhabha National Institute, Anushakti Nagar, Mumbai, India
3 Department of Biostatistics, Tata Memorial Centre, Paymaster Shodhika, ACTREC, Kharghar, Navi Mumbai, India


Correspondence
Dr. Navin Khattry, Professor and BMT Programme Co-ordinator, HSCT unit, Department of Medical Oncology, Room 211, Paymaster Shodhika, ACTREC, Kharghar, Navi Mumbai 410210, Maharashtra, India
Phone +91 989 2501 884
E mail: nkhattry@gmail.com

Engraftment fever (EF) is a common complication of autologous HSCT (AHSCT). It is difficult to discern it from infectious fever (IF). We studied the significance of total blood leucocyte count (TLC) to C-reactive protein (CRP) ratio in differentiating EF from IF. 109 consecutive AHSCT patients were retrospectively analysed between March 2011 and August 2013. Breakthrough fever (BF) was defined as new-onset fever preceded by an afebrile period of at least 48 hours. The BF episodes were classified as IF or EF. Infectious fever was diagnosed in case of blood culture positivity, radiological signs of infection, or fever subsiding within 48 hours of changing the antibiotics. Engraftment fever was defined in cases associated with rising leucocyte counts without identifiable infective focus. EF responded well to steroid therapy. Daily TLC and CRP values were obtained from patients’ records. Optimal cut-off value of ratio on day of BF was obtained by plotting ROC curve. Sensitivity and specificity were calculated at this value.

Among 109 cases, the breakthrough fever manifested in seventy patients. The median term for BF was day +9. Sixty-two patients had the EF. Median value of TLC/CRP ratio on the day of BF was significantly higher in patients with EF than with IF (0.139 vs 0.038, p=0.013). With ROC analysis, the AUC value was 0.78 (95%CI – 0.66-0.89, p<0.0001). The ROC curve provided the optimal TLC/CRP value of 0.056. Using a ratio >0.056 for EF, the sensitivity and specificity were 63% (95%CI 50-75%) and 100% (95%CI 63-100%) respectively. TLC/CRP ratio >0.056 is highly specific for EF. Prospective studies are warranted to confirm these findings.

Keywords

Total leukocyte count, C-reactive protein, engraftment fever, infectious fever, stem cell transplant, autologous.

Clinical studies

Bacteroides fragilis is a potential marker of effective microbiota transplantation in acute graft-versus-host disease treatment

Download PDF version

Oleg V. Goloshchapov1, Evgenyi A. Bakin1, Maxim A. Kucher1, Oksana V. Stanevich1, Maria A. Suvorova2, Vladimir V. Gostev3, Oleg S. Glotov4, Yury A. Eismont4, Dmitry E. Polev5, Anastasia Yu. Lobenskaya5, Ruslana V. Klementeva1, Maria O. Goloshchapova1, Ludmila S. Zubarovskaya1, Sergey V. Sidorenko3, Alexander N. Suvorov6, Ivan S. Moiseev1, Alexei B. Chukhlovin1

1 RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia
2 Explana Research Laboratory, St. Petersburg, Russia
3 Pediatric Research and Clinical Center of Infectious Diseases, St. Petersburg, Russia
4 City Hospital No. 40, St. Petersburg, Russia
5 Cerbalab Ltd, St. Petersburg, Russia
6 Institute of Experimental Medicine, St. Petersburg, Russia


Correspondence
Dr. Oleg V. Goloshchapov, Head, Anesthesiology Department No.3, Pavlov University, L. Tolstoy St. 6-8, 197022, St. Petersburg, Russia
Phone: + 7 (921) 979 2913
E-mail: golocht@yandex.ru

Fecal microbiota transplantation (FMT), as any other medical procedure, requires standardization of results, approaches, monitoring of its dynamics and microbiota engraftment evaluation. The aim of the present study was to compare efficiency and results of PCR and 16S RNA-based sequencing in order to trace the dynamics of microbiota composition after FMT.

Patients and methods

The prospective, single-center study included 27 patients with acute intestinal and chronic (overlap syndrome) graft-versus-host disease (GvHD) developed after allogeneic hematopoietic stem cell transplantation (HSCT). FMT in 19 cases was performed, mostly, with ingestible capsules, eight placebo-treated patients were included into control group. Quantitative changes of different bacterial groups in fecal microbiota were assessed by means of real-time multiplex PCR, being compared with16S rRNA sequencing technique at the terms of D+3, D+16, D+30, D+60 and D+120 following FMT. Clinical response was determined by 4 scales evaluating intestinal syndrome and GvHD grade.

Results

When evaluating stool consistence according to Bristol scale as an index of GvHD therapy efficiency, we have observed complete clinical response by the D+120 after FMT in nine cases (47% with Bristol score of ≤4 points), and nine patients (47%) showed improved stool properties (>4 points). In the placebo group, complete or partial response was revealed, respectively, in one (13%), and four cases (50%) on the D+120. Multiplex PCR of fecal microbiota has shown a different time course in FMT- and placebo-treated patients, when compared to their initial (pre-FMT) levels. Total bacterial mass and copy numbers of distinct microbial species exhibited sufficient increase after FMT. Such shifts were demonstrable on D+30 for total microbial mass (p=0.002); Escherichia coli (p=0.001); Bacteroides fragilis group (p=0.05); Faecalibacterium prausnitzii (p=0.005). Meanwhile, the numbers Lactobacillus spp., and Bacteroides thetaiotaomicron, generally, were not changed over this time period. Moreover, in the control group (placebo) we have not found significant fecal microbiota changes against initial levels during 120 days monitoring period. Over 120 days of observation, we have also found some differences of the microbiota dynamics for the subgroups with complete response and partial/no response: Bifidobacterium spp. (р<0.047), E.coli (р<0.00047), B. fragilis group (p=5.6×10-5), F.prausnitzii (р<0.0062).

Conclusions

1. Quantitative PCR of the major bacterial groups of gut microbiota, e.g., Bifidobacterium spp., E. coli, B. fragilis group, F. prausnitzii could be used as microbiological markers for evaluation of changing microbial landscape after FMT as a routine molecular biology technique. 2. The genocopy counts of B. fragilis group correlate with clinical response in the patients with intestinal GvHD after HSCT, either with, or without FMT procedure.

Keywords

Graft-versus-host disease, fecal microbiota, transplantation, gut microbiome, Bacteroides fragilis, polymerase chain reaction, multiplex, next generation sequencing, 16S rDNA gene.

Clinical studies

Morphological skin evaluation in the patients with systemic scleroderma before and after hematopoietic stem cell transplantation

Download PDF version

Galina V. Fedotovskikh, Galija M. Shaymardanova, Manarbek B. Askarov, Ainash A. Zhusupova, Natalya A. Krivoruchko, Tatyana G. Ezhelenko, Sapargul Marat

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


Correspondence
Prof. Galina V. Fedotovskikh, MD, National Scientific Medical Center, Abylaikhan Avenue 42, 010000, Nur-Sultan (Astana), Kazakhstan
Phone: +7 (707) 222 3256
E-mail: gvf_fedotovskikh@mail.ru

Some promising clinical results of hematopoietic stem cell transplantation (HSCT) are reported in severe autoimmune diseases. When treating the patients with systemic scleroderma (SSD), histological evaluation of skin fibrosis includes scoring of myofibroblasts that are associated with excessive deposition of extracellular matrix components. The aim of our study was to evaluate morphological condition of skin in SSD patients before and after transplantation of autologous hematopoietic bone marrow stem cells.

Patients and methods

Twenty-eight patients were observed at the National Research Medical Center (Nur-Sultan), at the age of 45+11 years old (2 males, 26 females) with verified diagnosis of SSD according to ACR/EULAR (2013). Duration of the disease was 12+5.4 years old. Control group (12 persons) received conventional basic therapy Treatment protocol for the main group included autologous HSCT. Resistance to immunosuppressive therapy was a pre-requisite for HSCT.

Bone marrow aspiration was performed from the iliac crest, the autologous mononuclear cells were isolated in Percoll density gradient and incubated for 72 hours at 37°С. Autologous HSCT was performed at a mean dose of 88×106 cells in 200 mL of physiological saline i/v over 3 hours. Clinical effect was evaluated by the recognized criteria (European Scleroderma Trials and Research Group, skin score by Rodnan). For morphological studies, the punch biopsies of tibial skin were taken in 15 patients before therapy, and in 9 three months after the treatment. The cellular therapy was accompanied by improved skin condition. The paraffin sections were stained with hematoxilin and eosin, as well as by Masson-trichrome technique. For electron microscopy, the skin biopsies were processed by conventional method, then being Epon-embedded. Semi-thin slices were stained with Methylene Blue, Azur II and basic fuchsine. For EM, the ultrathin sections were contrasted with uranyl acetate and lead citrate.

Results

Skin of SSD patients before treatment was characterized by induration and dystrophy and epithelial destruction, sclerosis and hyalinosis of dermal connective tissue, pathology of microcirculatory vessels. Ultrastructure of myofibroblasts in the sclerotized derma was characterized by functional overload. The active participants of fibrillogenesis were located in perivascular area, being represented by lymphocytes and fibroblasts of a specific SSD-specific population producing higher amounts of collagen and interstitial matrix. However, three months after HSCT, the main group of the patients exhibited a pronounced clinical effect with sufficient decrease of skin induration, reduced dysphagia, mitigation of muscle contractures, couping vasospasm attacks (Raynaud syndrome). Skin density was significantly decreased, with Rodnan scores changed from 12.9 to 8.7 (only 1-point decrease in the controls). HSCT promoted biodegradation of skin fibrotic tissue in SSD patients. The myofibroblasts were subjected to destruction, multiple and prolonged capillaries were observed, the cell composition of perivascular infiltrate shifted to normal state. Numerous phagocytic and secretory macrophages appeared, thus suggesting angiogenesis induction, tissue remodeling, regulation of fibroclast population, suppression of T- and B-lymphocytes playing an important role on SSD pathogenesis. Extensive telocyte connections presumed their participation in neoangiogenesis and transmission of regeneration signaling.

Conclusion

Transplantation of cultured autologous hematopoietic marrow stem cells in SSD patients promoted biodegradation of sclerotized dermal layer, as well as angiogenesis stimulation, restoration of epithelium and skin appendages 3 months after HSCT, thus corresponding to improvement of clinical symptoms.

Keywords

Systemic scleroderma, hematopoietic stem cell transplantation, bone marrow, skin morphology.

Clinical case

A long-term response to allogeneic hemopoietic stem cell transplantation from haploidentical donor and post-transplant therapy in an adolescent with primary resistant neuroblastoma

Download PDF version

Ilya V. Kazantsev1, Tatiana V. Iukhta1, Asmik G. Gevorgian1, Polina S. Tolkunova1, Andrew V. Shamin2, Vadim V. Baykov3, Nikolay A. Vorobyov4, Andrew V. Kozlov1, Marina A. Karsakova2, Polina S. Kuga1, Alexander N. Shvetsov1, Elena V. Morozova1, Svetlana S. Safonova1, Yuri A. Punanov1, Ludmila S. Zubarovskaya1, Boris V. Afanasyev1

1 RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia
2 N. N. Ivanova Samara Regional Pediatric Hospital, Samara, Russia
3 Chair of Pathology, Pavlov University, St. Petersburg, Russia
4 Dr. Berezin Medical Institute, St. Petersburg, Russia; I. Mechnikov North-Western State Medical University; Saint Petersburg State University, St. Petersburg, Russia
5 Department and Chair of Roentgenology, Pavlov University, St. Petersburg, Russia


Correspondence
Dr. Ilya V. Kazantsev, RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University,
L. Tolstoy St. 6-8, 197022, St. Petersburg, Russia
Phone: +7 (963) 348 0524
E-mail: Ilya_Kazantsev@inbox.ru

Neuroblastoma (NB) is the most frequent pediatric extracranial solid tumor characterized by extreme biological heterogeneity with variable clinical course. Older age is an important risk factor. These patients may lack other common risk features but still have a chemoresistant disease with dismal prognosis. As there is currently no consensus on optimal treatment for patients with primary resistant NB, a number of clinical options is being explored including immunotherapy-based approaches. Immunotherapy with dinutuximab beta (DB) have proven its effectiveness as maintenance therapy. Allogeneic stem cell transplantation from haploidentical donor (haplo-HSCT) may be an effective consolidation in some cases. However, all forms of immunotherapy are much less effective in patients with large residual tumor. While there is no data on immune checkpoints inhibitors effectiveness in NB, some patients may benefit from this option as a part of complex immunotherapy strategy.

Case presentation

A 12-year old girl with gross paravertebral thoracic and abdominal tumor was diagnosed with undifferentiated neuroblastoma and bone metastases. While there was no response to several lines of chemotherapy, and only partial tumor resection was possible, the hematopoietic stem cell transplantation from haploidentical donor (haplo-HSCT) was performed as salvage therapy. Since there was only minor decrease in tumor volume with good dynamics by MIBG scan, additional post-transplant therapy was initiated. External beam radiotherapy was given for local control. The patient also received combined immunotherapy with DB and nivolumab. Currently, 3.5 years post haplo-HSCT, despite still gross residual tumor mass, it is MIBG-negative and shows signs of differentiation.

Conclusion

The combination of haplo-HSCT with post-transplant anti-GD2 and nivolumab may lead to a long-term response in an adolescent with primary resistant NB in spite of a large residual tumor mass.

Keywords

Neuroblastoma, pediatric, resistant, hematopoietic transplantation, immunotherapy.

Clinical case

Successful treatment of relapsed/refractory B-Acute lymphoblastic leukemia with Inotuzumab ozogamicin after blinatumomab failure

Download PDF version

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

RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia


Correspondence
Dr. Sergey N. Bondarenko, RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, L. Tolstoy St. 6-8, 197022, St. Petersburg, Russia
Phone: +7 (812) 338 62372
E-mail: dr.sergeybondarenko@gmail.com

Blinatumomab, a bispecific T-cell engaging CD3-CD19 antibody, is highly effective in patients with relapsed/refractory (R/R) B-cell acute lymphoblastic leukemia (ALL) even after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, patients who failed with Blina have a dismal outcome. Inotuzumab ozogamicin is one of the therapeutic options after blinatumomab failure. We report a young man who exhibited bone marrow (BM) relapse of B-ALL following haploidentical stem cell transplantation (haplo-HSCT). Remission was not achieved after Blinotumomab treatment, thus Inotuzumab was administered. A complete remission with no signs of minimal residual disease was achieved after a single cycle of Inotuzumab. The second haplo-HSCT from another donor was successful.

Conclusion

The present case demonstrate an opportunity of successful inotuzumab therapy after failure of allo-HSCT and blinotumomab treatment.

Keywords

Hematopoietic stem cell transplantation, allogeneic, acute lymphoblastic leukemia, Blinotumomab, Inotuzumab ozogamicin, relapsed/refractory.

Experimental studies

Augmented reality technology for auricular reconstruction in the treatment of microtia

Download PDF version

Аndrey I. Yaremenko1, Anna V. Lysenko1, Elizaveta A. Ivanova1, Oleg V. Galibin2

1 Department of Maxillofacial Surgery, Pavlov University, St. Petersburg, Russia
2 RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia


Correspondence
Dr. Elizaveta A. Ivanova, Department of Maxillofacial Surgery, Pavlov University, L. Tolstoy St. 6-8, 197022, St. Petersburg, Russia
Phone: +7 (953) 144 1508
E-mail: lizabet159@yandex.ru

Facial defects and deformations occupy a significant place in the practice of maxillofacial and dental surgeons. Nevertheless, maxillofacial surgery is developing rapidly and requires improvement of existing treatment methods, and introduction of new approaches to reconstructive surgery. Augmented reality is a promising direction of computer technology development which is actively used in medicine and education. Modern computer technology allows to create a 3D model of a lost organ and use it for preoperative planning, as well as apply a virtual model for intraoperative navigation. Recently, the method of augmented reality has been actively developed, when a virtual image of the zone of the surgical area or a dedicated organ is used, which is compared with its real prototype in static mode, or in real-time using computer devices. The benefits of using augmented reality technologies in reconstructive surgery is associated with preoperative virtual planning, simplification of the surgical intervention itself, as well as with a reduction in the risks of intra- and postoperative complications. The aim of our work was to study the opportunity of using the augmented reality technology in reconstructive surgery for microtia correction based on pre-operative computer simulation.

At the preoperative stage, a photometric analysis of the patient was carried out, then a computer simulation of the missing auricle was performed. Using a 3D printer, a virtual model of the reconstructed auricle was obtained. The image in three-dimensional format was loaded into augmented reality glasses, which made it possible to project the shape and position of the simulated auricle to the area of the defect of the auricle when preparing for surgery. During the surgery, a marker was installed near the surgical field, in order to display the three-dimensional model in a destined position. During surgical intervention, an autogenous costal cartilage was taken, from which the auricle was formed using augmented reality approach and three-dimensional modeling. Subsequently, the graft was introduced to the formed bed in the area of the right ear auricle.

The obtained 3D model of the auricle before the operation enabled planning of the forthcoming operation and determine the amount of autograft needed for reconstruction. Using the augmented reality glasses, the exact shape of the auricle is reproduced during the operation, and its proper position is assessed in relation to the healthy side. No complications were observed over the postoperative period.

Virtual modelling of a lost or absent organ based on a preoperative examination provides important information about its spatial structure. Preoperative virtual planning allows you to predict the individual features of the operation, its difficult stages, to anticipate possible complications. The use of augmented reality technology during reconstructive surgery is a promising method requiring further development and improvement.

Keywords

Ear, microtia, cartilage, transplantation, augmented reality, 3D printing.

Obituary

In memory of Professor Rolf Neth October 6, 1926 – March 17, 2020

Download PDF version

Boris Fehse, Nicolaus Kröger, Carol Stocking, Axel Zander

With great sadness, we learned of the passing of Professor Dr. med. Rolf-Dietmar Neth, the founder of the Wilsede meeting, on March 17, 2020, aged 93 in his home town Buchholz near Wilsede/Lüneburger Heide. Rolf Neth was born 6.10.1926. After the 2nd World War, Rolf Neth studied Medicine from 1949 to 1955 at the University of Göttingen. Then he was at the Max-Planck-Institut für Experimentelle Medizin (University Göttingen, 1956-1957), and promoted his skills in clinical and experimental hematology in St. George Hospital (1958-1959) in Hamburg.

Fehse-fig01.jpg

From 1960, his activity was connected with the pediatric clinics of Hamburg University where he became a Professor at the Children Hospital in 1972. From 1970 to 1980, Rolf Neth was occupied implementing new laboratory diagnostic approaches in the booming field of clinical hematology. Blood cancer treatment was developed, due to novel drugs invented to combat leukemic cells and rescue the small patients which 10-20 years ago had only zero chance to survive. Histo- and immunochemical diagnostics became routine tests for evaluation of clinical forms of leukemias and efficiency of their therapy. Since 1982, he coordinated laboratory hematology at the Department of Clinical Chemistry, University Hamburg, until retirement in 1992.

Along with contribution to clinical laboratory science, Professor Neth, over 1973 to 2002, arranged a series of famous Wilsede Meetings "Modern Trends in Human Leukemia" dedicated to leukemia research and treatment. Rolf Neth and Robert Gallo decided time and topic of the meeting, and Rolf Neth proposed a place, i.e., a lonely village in the Luneburg heath, not far from his home. Hence, Leukemia and Viruses was selected as a specific topic for a meeting in Germany, because it was timely for convergence between clinical medicine and cancer biology.

Rolf Neth organized the first Wilsede meetings himself for more than 20 years, until he passed these efforts to Wolfram Ostertag and Axel Zander. In the late 1990s, Carol Stocking and Boris Fehse took on this responsibility. Over last years, Wilsede meetings were arranged by Nicolaus Kröger and Boris Fehse. And as long as his health permitted, Rolf Neth came along to see how his baby was doing. He participated and assisted at any stage of the next meeting. We are thankful to have had the privilege of knowing and cooperating with Rolf Neth and to cherish his legacy by keeping the Wilsede tradition alive. He was married with Hanne-Lore Cohrs, 8.11. 1958, survived by his wife of 62 years Hanne-Lore, and four sons and several grandchildren.

References

  1. R. Neth, G. Schwarting. Das Verhalten der Koronarsklerose in der Nachkriegszeit. Dtsch.med.Wschr. 80,570-573 (1955).
  2. R. Neth. Die diagnostische Bedeutung cyto- und mikrochemischer Eisenstoffwechseluntersuchungen" Klin.Wschr. 44, 687-695 (1966).
  3. R. Neth. Catalysis of Peptidyl Transfer by human tonsil ribosomes and effects of some antibiotics. FEBS Letters 8, 198-202 (1970).
  4. R.D. Neth. Blutbild und Urinstatus. Springer-Verlag Berlin Heidelberg GmbH 1979.
  5. Modern Trends in Human Leukemia I Gruner + Stratton, J.F.Lehmannverlag 1974, II-IX 1976-1992, Springerverlag in science-connections.com
  6. Hanne-Lore Neth. "Heidepastor" Wilhelm Bode. Hämatol. Bluttransf. Vol 35.

Obituary

Rolf Neth and Russia

Download PDF version

Prof. Margarita B. Belogurova,
Department of Pediatric Oncology, City Hospital No. 31, St. Petersburg, Russia

Prof. Ludmila S. Zubarovskaya,
RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia

Rolf Neth was also eager to keep scientific borders open and established early connections with scientists from Eastern Europe, particularly the Soviet Union, during the cold war. After the Chernobyl accident (April 1986) a number of doctors and scientists (e.g., R. P. Gale and P. Terasaki) arrived to Moscow within several days, in order to assist Russian doctors by attempting bone marrow transplantation in severely irradiated patients. To study long-term effects of Chernobyl accident, the affected areas of Ukraine and Belarus required well-arranged medical surveillance which was established by international collaboration of European scientists. Professor Rolf Neth has undertaken great efforts to develop appropriate medical infrastructure in radioactively contaminated areas and to organize appropriate laboratory facilities, mostly, for pediatric healthcare (Fig. 1).

Belogurova-fig01.jpg

Figure 1. Prof. Rolf Neth, Prof. A. I.Vorobyov and leading Russian hematologists visiting Chernobyl accident area (1989)

In addition to coordinating essential medical measures after the Chernobyl accident, Rolf Neth has organized, together with Elena I. Frolova (Fig. 2, left), the first Volga-Wilsede Meeting in 1990, at the edge of USSR fall. It is regrettable that Professor Boris Afanasyev (Saint Petersburg) who arranged the successive Wilsede-Neva meeting (Fig. 2, 3) died on March 16, 2020, one day before Rolf Neth.

Belogurova-fig02.jpg

Figure 2. Discussing an idea of joint Wilsede Meeting on the Volga June 23 1988 in Buchholz. Elena Frolova, Joseph Chertkov, Elena Elsner, Hanne-Lore Neth, Rolf Neth, Alexander Friedenstein, Boris Afanasyev

Belogurova-fig03.jpg

Figure 3. Prof. Rolf Neth (right) and Prof. Boris Afanasyev (Left), 1991. Kindly provided by O. F. Afanasyeva

The joint German-Russian Wolga-Wilsede Meeting (1990) was the first experience of such Symposium in Russia. It was arranged as a ship travel, and we had our first experience in this field, arranging the program, preparing posters and oral reports. For the first time we attended research communications in the cabin company or ship restaurant, alternated by short anchorages at the Volga strands, accompanied by joyful parties with our young colleagues, with friendly attitude of our older Masters. From Russian side, it was arranged by Elena Frolova, Gregory Dolganov and a number of Moscow hematologists.

Another Wilsede Meeting was scheduled in Saint Petersburg as the Neva-Wilsede Symposium (1992). Its guests arrived to Saint Petersburg, then boarded the tourist ship, then went across Ladoga and Onega lakes, observing the Russian Valaam and Kizhi monuments, and picturesque landscapes underway. The scientific program was as interesting as social events. This meeting included a special symposium on radiation biology, moderated by Prof. Gerard Wagemaker which concerned medical consequences of Chernobyl accident.

During the Wilsede Meeting 1990, Prof. Boris V. Afanasyev has met several leading professors from Hamburg University: R. Neth, A. Zander, K. Winkler, G. Janka. Subsequently, with support of Hamburg Major Henning Voscherau they have adopted a program of training medical staff in bone marrow transplantation (Prof. A. Zander) and pediatric leukemia treatment (Prof. K. Winkler, G. Janka), as well as modern diagnostic techniques (Prof. R. Neth) in the frame of collaboration between Hamburg and Leningrad, the two partner cities. Since 1990, about 100 doctors and medical nurses were trained in Hamburg University in chemotherapy of blood cancer and solid malignancies in children and adults, bone marrow transplantation, blood transfusion, molecular biology, tissue typing.

Prof. Margarita B. Belogurova remembers: during our first training of in Hamburg-Eppendorf University Clinics (1991), Professor Axel R. Zander introduced me to Prof. Rolf Neth who arranged training programs for Russian doctors. Previously, these contacts were only occasional, since Neth was not a clinician, but this time he invited us to his home together with other Russian-speaking doctors, I guess, from Ukraine. It was a cosy house, a dinner prepared by his wonderful wife, and long conversations or talks about different things: our desire for better diagnostics and treatment of leukemia (as in Germany), about our needs and requirements, and potential indispensable aid for Russian medicine. Russian-speaking guests heard some words and phrases in Russian from Rolf Neth, and listened to his personal story of his military service during the World War II, severe frostbites and imprisonment in Russia. Since that time, he had severe injuries of feet thus requiring orthopedic foorwear. He described these dramatic episodes as usual life events! He was devoid of any snobbism, showing warmth and cordiality to Russians.

After organization of BMT Center in the Saint Petersburg City Hospital No.31, its chief, the late Professor Boris Afanasyev has involved all the staff into clinical and fundamental research. Being invited to the Wilsede scientific meetings, we, as ordinary audience, were amazed by unusual looks to the old German village, however, well packed with modern conference halls and equipment. International audience and foreign environment were also surprising for us. Due to great support of Prof. Axel Zander, Rolf Neth, Gritta Janka, Boris Fehse, many Russian doctors became an excellent opportunity of research-oriented clinical training in pediatric oncology, oncohematology and relevant laboratory studies in the clinical and laboratories of Hamburg University.

When we remember Professor Rolf Neth, the simple formula of success is kept in mind: the long way begins with first step. Likewise, the story of noble German help for Russia followed similar way: From humanitarian aid towards the collaboration of equal partners.