MSCs products as well as their derived extracellular vesicles, are currently being explored as advanced biologics in cell-based therapies with high expectations for their clinical use in the next few years. In recent years, various strategies designed for improving the therapeutic potential of mesenchymal stromal cells (MSCs), including pre-conditioning for enhanced cytokine production, improved cell homing and strengthening of immunomodulatory properties, have been developed but the manufacture and handling of these cells for their use as advanced therapy medicinal products (ATMPs) remains insufficiently studied, and available data are mainly related to non-industrial processes. In the present article, we will review this topic, analyzing current information on the specific regulations, the selection of living donors as well as MSCs from different sources (bone marrow, adipose tissue, umbilical cord, etc.), in-process quality controls for ensuring cell efficiency and safety during all stages of the manual and automatic (bioreactors) manufacturing process, including cryopreservation, the use of cell banks, handling medicines, transport systems of ATMPs, among other related aspects, according to European and US legislation. Our aim is to provide a guide for a better, homogeneous manufacturing of therapeutic cellular products with special reference to MSCs.
To investigate the three-dimensional (3D) genome architecture across normal B cell differentiation and in neoplastic cells from different subtypes of chronic lymphocytic leukemia and mantle cell lymphoma patients, here we integrate in situ Hi-C and nine additional omics layers. Beyond conventional active (A) and inactive (B) compartments, we uncover a highly-dynamic intermediate compartment enriched in poised and polycomb-repressed chromatin. During B cell development, 28% of the compartments change, mostly involving a widespread chromatin activation from naive to germinal center B cells and a reversal to the naive state upon further maturation into memory B cells. B cell neoplasms are characterized by both entity and subtype-specific alterations in 3D genome organization, including large chromatin blocks spanning key disease-specific genes. This study indicates that 3D genome interactions are extensively modulated during normal B cell differentiation and that the genome of B cell neoplasias acquires a tumor-specific 3D genome architecture. ; This research was funded by the European Union's Seventh Framework Programme through the Blueprint Consortium (grant agreement 282510), the World Wide Cancer Research Foundation Grant No. 16-1285 (to J.I.M.-S.), the ERC (grant agreement 609989 to M.A.M.-R.), European Union's Horizon 2020 research and innovation programme (grant agreement 676556 to M.A.M.-R.). We also knowledge the support of Spanish Ministerio de Ciencia, Innovación y Universidades through SAF2012-31138 and SAF2017-86126-R to J.I.M.-S., SAF2015-64885-R to E.C., BFU2017-85926-P to M.A.M.-R. and PMP15/00007 to E.C. which is part of Plan Nacional de I + D + I and co-financed by the ISCIII-Sub-Directorate General for Evaluation and the European Regional Development Fund (FEDER-"Una manera de Hacer Europa") (to E.C.), the International Cancer Genome Consortium (Chronic Lymphocytic Leukemia Genome consortium to E.C.), La Caixa Foundation (CLLEvolution-HE17-00221, to E.C.). Furthermore, the authors would like to thank the support of the Generalitat de Catalunya Suport Grups de Recerca AGAUR 2017-SGR-736 (to J.I.M.-S.), 2017-SGR-1142 (to E.C.) and 2017-SGR-468 (to E.C.), the Accelerator award CRUK/AIRC/AECC joint funder-partnership, the CERCA Programme/Generalitat de Catalunya and CIBERONC (CB16/12/00225, CB16/12/00334, and CB16/12/00489). R.V.-B. (BES-2013-064328) and P.S.-V. (BES-2014-070327) were supported by a predoctoral FPI Fellowship from the Spanish Government. CRG acknowledges support from 'Centro de Excelencia Severo Ochoa 2013-2017', SEV-2012-0208 and the CERCA Programme/Generalitat de Catalunya as well as support of the Spanish Ministry of Science and Innovation through the Instituto de Salud Carlos III and the EMBL partnership, the Generalitat de Catalunya through Departament de Salut and Departament d'Empresa i Coneixement, and the Cofinancing with funds from the European Regional Development Fund (ERDF) by the Spanish Ministry of Science and Innovation coresponding to the Programa Opertaivo FEDER Plurirregional de España (POPE) 2014-2020 and by the Secretaria d'Universitats i Recerca, Departament d'Empresa i Coneixement of the Generalitat de Catalunya corresponding to the programa Operatiu FEDER Catalunya 2014-2020
Understanding the regulation of normal and malignant human hematopoiesis requires comprehensive cell atlas of the hematopoietic stem cell (HSC) regulatory microenvironment. Here, we develop a tailored bioinformatic pipeline to integrate public and proprietary single-cell RNA sequencing (scRNA-seq) datasets. As a result, we robustly identify for the first time 14 intermediate cell states and 11 stages of differentiation in the endothelial and mesenchymal BM compartments, respectively. Our data provide the most comprehensive description to date of the murine HSC-regulatory microenvironment and suggest a higher level of specialization of the cellular circuits than previously anticipated. Furthermore, this deep characterization allows inferring conserved features in human, suggesting that the layers of microenvironmental regulation of hematopoiesis may also be shared between species. Our resource and methodology is a stepping-stone toward a comprehensive cell atlas of the BM microenvironment. ; We would like to thank the staff of the flow cytometry core, the advances genomic lab, and the animal facility at CIMA Universidad de Navarra for their invaluable technical and intellectual assistance. We are particularly grateful to the healthy volunteers who donated bone marrow tissue for this study. We also acknowledge Ali O. Balubaid's help in writing. We would like to acknowledge Miguel Cocera-Fernandez's contribution to the Graphical Abstract. ; Funded by grants from The Spanish Government, through project PID2019-111192GA-I00 (MICINN) to DGC. Instituto de Salud Carlos III (ISCIII) and co-financed by FEDER: PI16/02024, PI17/00701 and PI19/01352, TRANSCAN EPICA AC16/00041, CIBERONC CB16/12/00489; Redes de Investigación Cooperativa (TERCEL RD16/0011/0005); Spanish Ministry of Economy, Industry and Competitivity (RTHALMY SAF2017-92632-EXP); Departamento de Salud, Gobierno de Navarra 40/2016 and Departamento de Desarrollo Económico y Empresarial (AGATA 0011-1411-2020-000010 and 0011-1411-2020-000013). The study was also ...
Introduction Based on the advances in the treatment of multiple sclerosis (MS), currently available disease-modifying treatments (DMT) have positively influenced the disease course of MS. However, the efficacy of DMT is highly variable and increasing treatment efficacy comes with a more severe risk profile. Hence, the unmet need for safer and more selective treatments remains. Specifically restoring immune tolerance towards myelin antigens may provide an attractive alternative. In this respect, antigen-specific tolerisation with autologous tolerogenic dendritic cells (tolDC) is a promising approach. Methods and analysis Here, we will evaluate the clinical use of tolDC in a well-defined population of MS patients in two phase I clinical trials. In doing so, we aim to compare two ways of tolDC administration, namely intradermal and intranodal. The cells will be injected at consecutive intervals in three cohorts receiving incremental doses of tolDC, according to a best-of-five design. The primary objective is to assess the safety and feasibility of tolDC administration. For safety, the number of adverse events including MRI and clinical outcomes will be assessed. For feasibility, successful production of tolDC will be determined. Secondary endpoints include clinical and MRI outcome measures. The patients' immune profile will be assessed to find presumptive evidence for a tolerogenic effect in vivo. Ethics and dissemination Ethics approval was obtained for the two phase I clinical trials. The results of the trials will be disseminated in a peer-reviewed journal, at scientific conferences and to patient associations. ; A FACTT network (Cost Action) [BM1305]; EU Framework Program Horizon 2020; European Union's Horizon 2020 research and innovation program [779316]; applied biomedical research project of the Institute for the Promotion of Innovation by Science and Technology in Flanders [IWT-TBM 140191]; Platform for Clinical Research and Clinical Trial Units, Spanish Clinical Research Network, SCReN [PI11/02416, PI14/01175, PI16/01737, PT13/0002/0038]; Health Institute Carlos III -Subdireccion General de Evaluacion y Fomento de la Investigacion of the Spanish Ministry of Economy and Competitiveness; Fondo Europeo de Desarrollo Regional (FEDER)European Union (EU); Fundacio La Marato de TV3 [07/2410]; Sanofi Genzyme, Belgium; Research Foundation Flanders (FWO)FWO; Hospital Germans Trias i Pujol ('Germans Trias Talents 2016-2018'); University of Antwerp; Research Foundation FlandersFWO [FWO 1701919N]; Spanish Patient association 'Treball de Vida' (Associacio d'Afectats d'Esclerosi Multiple del Barcelones Nord i Maresme)
