The 44Th Annual Meeting of the European Society for Blood and Marrow Transplantation: Physicians Award Winners
Total Page:16
File Type:pdf, Size:1020Kb
Bone Marrow Transplantation (2019) 53:13–18 https://doi.org/10.1038/s41409-018-0317-z ABSTRACT The 44th Annual Meeting of the European Society for Blood and Marrow Transplantation: Physicians Award Winners 18–21 March 2018 ● Lisbon, Portugal Published online: 24 September 2018 © Springer Nature Limited 2018 Copyright: Modified and published with permission from http://www.ebmt2018.org/ Sponsorship Statement: Publication of this supplement is sponsored by the European Society for Blood and Marrow Transplantation. PHYSICIANS AWARD WINNERS in neurologic disability. We now report results on a randomized trial of non-myeloablative HSCT versus 1234567890();,: 1234567890();,: Van Bekkum Award continued treatment with standard DMTs. O001 Methods: Patients on stable disease modifying therapy (DMT) with > 2 relapses in the previous 12 months Non-myeloablative haematopoietic stem cell were randomized (1:1) to treatment with either cyclopho- transplantation versus continued disease modifying sphamide and rabbit anti-thymocyte globulin followed therapies (DMT) in patients with highly active relapsing by hematopoietic stem cell infusion or to a control arm remitting multiple sclerosis (RRMS) with continued treatment with standard DMTs. Evaluating neurologists scoring the Expanded Disability Status Richard K Burt1, Roumen Balabanov2, John A Snowden3, Scale (EDSS) were blinded to treatment arms. Patients Basil Sharrack4, Maria Carolina Oliveira5, Flavia Nelson6, in the control arm who had 6 month confirmed EDSS Joachim Burman7 increase of > 1 point despite at least one year of treatment 1Northwestern University, Division of Immunotherapy for (defined as treatment failure) were allowed to crossover Autoimmune Diseases, Chicago, IL, United States; to HSCT. 2Northwestern University, Department of Neurology, Chi- cago, IL, United States; 3Sheffield Teaching Hospitals NHS Results: 110 patients were randomized, 55 to each arm. Foundation Trust, Department of Haematology, Sheffield, Three HSCT patients were withdrawn: two for failing United Kingdom; 4Sheffield Teaching Hospitals NHS enrollment criteria, one for recurrent infections occurring Foundation Trust, Department of Neurology, Sheffield, before transplant. Five control patients were withdrawn United Kingdom; 5University of São Paulo, Department of after soliciting transplants at other centers. All patients Internal Medicine, São Paulo, Brazil; 6University of Texas are at least one-year post enrollment. No deaths occurred Health Science Center at Houston, Department of Neurol- and no CTC grade 4 non-hematopoietic toxicities ogy, Houston, TX, United States; 7Uppsala University, occurred in the transplant arm. DMTs (number of patients) Department of Neuroscience, Uppsala, Sweden used in the control arm were: natalizumab (22), dimethyl fumarate (18), fingolimod (13) interferons (10), glatiramer Background: We previously reported (Journal of the acetate (8), mitoxantrone (5). During the first year American Medical Association, 2015) that non- after enrollment, one relapse occurred on the HSCT myeloablative haematopoietic stem cell transplantation arm versus 39 on the DMT arm (P< 0.001). Mean (HSCT) may be performed safely in patients with multiple EDSS improved from 3.5 to 2.4 after HSCT while it wor- sclerosis and is accompanied by long-term improvement sened from 3.3 to 3.9 on DMTs (P< 0.001). With a 14 mean follow up of 3 years (range 1 to 5 years), treatment (NTA). VEs stained with Vibrant Dil were incubated with failure was 60% (30 of 50) for the control arm and 6% CD34+ cells from cord blood for 24 hours. Then their (3 of 52) for HSCT (P < 0.001). For the 30 patients incorporation into HPC was analyzed by flow cytometry who failed the control arm and crossed over to HSCT, by and confocal microscopy. To study gene expression chan- one year after HSCT, the mean EDSS improved from 5.2 ges elicited in HPC after MSC-EVs incorporation, expres- to 2.6 (P< 0.001). sion arrays were performed using the Human Gene 2.0ST platform (Affymetrix), evaluating the functions of the Conclusions: HSCT was statistically superior to continued involved genes with the Webgestalt software. Apoptosis DMTs in patients with RRMS with > 2 relapses a year was evaluated by flow cytometry using annexinV/7AAD staining, whereas cell cycle was analyzed with BD Cycle- Clinical Trial Registry: NCT00273364, https://clinicaltria testPlus DNA Reagent kit. In addition, clonal growth of ls.gov/ct2/show/NCT00273364?term=NCT00273364&ra HPCs was compared by clonogenic assays in methylcellu- nk=1 lose. Finally, the functional in vivo changes induced by MSC-EV incorporation were studied in a xenotransplanta- Conflict of interest: All the authors have nothing to tion animal model. For this purpose, control HPC or HPC disclose previously co-cultured with MSC-VEs were administered into the BM of NOD-SCID mice. After four weeks, human hematopoietic engraftment was determined by flow Basic Science Award cytometry. O002 Results: MSC-VEs were positive for both MSC (CD44, CD90) and VEs (CD81, CD63) markers and negative The incorporation of extracellular vesicles from for hematopoietic markers (CD34, CD45), and exhibited mesenchymal stromal cells into hematopoetic progenitors their characteristic morphology by electron microcopy increase their clonogenic capacity and their engraftment and a regular size between 50 and 200nm by NTA. ability The incorporation of EVs into HPC was verified both by confocal microscopy and flow cytometry. In the Silvia Preciado, Sandra Muntión, Ana Rico, Luis Antonio gene expression array analysis we found that MSC- Corchete, Ana Gómez de la Torre, Teresa L Ramos, Con- EV incorporation induced an overexpression of some cepción Rodríguez, Rebeca Ortega, Jorge Diego Rufino, anti-apoptotic genes (BIRC2, BIRC3, NFKB) and a Miriam López-Parra, Alba Redondo, Lika Osugui, Fermín downregulation of pro-apoptotic genes (caspases) with Sánchez-Guijo an overexpression of genes involved in colony formation Hospital Universitario de Salamanca, Salamanca, Spain and hematopoetic cell proliferation (CD22, IL3RA, IL11), together with an activation of genes of the JAK-STAT Background: Treatment of graft failure after bone marrow pathway and an overexpression of CD44. All these (BM) transplantation with mesenchymal stromal cells findings were confirmed by other methods at the gene (MSC) is being evaluated clinically, since preclinical or protein level. A significant decrease in apoptosis evidence shows that MSC co-transplantation increases (p = 0.031), an increase of the percentage of cells in phase S hematopoietic progenitor cell (HPC) engraftment in xeno- (p = 0.065) and an increased CD44 expression (p = 0.03) trasplantation models. Since MSC exert their functions, were confirmed by flow cytometry in HPC that had in part, by the release of extracellular vesicles (EVs), in incorporated MSC-EVs compared to control HPCs. In the current study we wanted firstly to analyze multi- addition, these HPCs displayed a higher CFU-GM clono- parametrically changes induced in HPCs after the incor- genic potential. Finally, in the xenotransplantation poration of EVs released from MSC (MSC-EVs) and model, the engraftment ability of human HPC co-cultured secondly compare their capacity of engraftment in vivo to with MSC-EVs was significantly higher 4 weeks that of control HPC. after transplantation (10.26% (0.31%–71.06%) vs 1.22% (0.15%–55.97%); p = 0.0078). Methods: BM-MSC were isolated from 10 healthy donors following standard methods and characterized following Conclusions: The incorporation of EVs derived from MSC ISCT criteria. MSC-EVs were isolated by ultracentrifuga- induces genomic and functional changes in HPC, increasing tion and characterized by flow cytometry, Western blot, their clonogenic capacity in vitro and their engraftment electron microscopy and nanoparticle tracking analysis ability in vivo. The 44th Annual Meeting of the European Society for Blood and Marrow. 15 Funding: GRS1348/A/16, PI12/01775 (ISCIII, Spain). Results: WES yielded a uniform and consistent coverage of Santander-USAL grant to SP all samples, with an average depth-of-sequencing of 120x for purified AML samples and 60X for donor and patient Conflict of interest: There is no conflict of interest. The germline controls. A total of 249 protein-damaging somatic authors declare no competing financial interests. mutations in 220 genes were detected among all leukemic samples, averaging 15 somatic mutations per sample. Of all mutations, 54 were exclusive to relapses, with the muta- Presidential Symposium tional burden increasing significantly from pre- to post- alloHSCT samples (Wilcoxon test, pvalue = 0.02). This O003 was reflected also by patterns of clonal evolution observed in our cohort, with emergence in 8 patients of new clones at Abstract previously published relapse. Of notice, both number of post-transplantation relapse-specific mutations and appearance of new clones O004 had a tendency to increase proportionally with time to relapse. Relapse-specific mutations encompassed known Genomic and Transcriptional Profiling of Acute Myeloid AML driver genes, including WT1 and KRAS, but no gene Leukemia by Next-Generation Sequencing Unravels evidently related to immune function. Patient-Specific Patterns of Post-Transplantation Relapse By a linear model analysis of RNA-Seq data we found ~500 genes significantly deregulated in leukemic cells at Lucia Zanotti1, Gabriele Bucci1,2, Francesco Santaniello1,2, post-transplantation relapse and, in stark contrast to the Cristina Toffalori1, Donatella Biancolini2,