Biobank quality management in the BBMRI.be network

From as early as 2005, different guidelines and quality standards covering biobank activities and sample handling methods have been developed to improve and guarantee the reproducibility of biomarker research. Ten years on, the BBMRI.be Quality working group wanted to gauge the current situation of these aspects in the biobanks of the BBMRI.be network. To this end, two online surveys were launched (fall 2017 and fall 2018) to the biobank quality managers in the BBMRI.be network to determine the status and setup of their current quality management system (QMS) and how their QMS and related practices have evolved over a 14 month time period. All biobanks addressed by the two surveys provided a complete response (12 and 13, respectively). A QMS was implemented in 85% of biobanks, with 4 standards emerging as primary basis. Supplementary guidelines were used, with a strong preference for the ISBER best practices for biobanks. The Standard Preanalytical Code-an indicator of the preanalytical lifecycle of a biospecimen impacting the downstream analysis results-was already implemented in 50% of the biobanks while the other half intends future implementation. To assess and maintain the quality of their QMS, 62% of biobanks used self-assessment tools and 71% participated in proficiency testing schemes. The majority of biobanks had implemented procedures for general and biobank specific activities. However, policies regarding the business and sustainability aspect of biobank were only implemented in a limited number of biobanks. A clear desire for a peer-review audit was expressed by 69% of biobanks, with over half of them intending to implement the recently published biobank standard ISO20387. Overall, the biobanks of the BBMRI.be network have actively implemented a solid quality approach in their practices. The implementation of ISO 20387 may bring further professionalization of activities. Based on the needs expressed in this survey, the Quality working group will be setting up an audit program for the BBMRI.be biobanks, to enhance, harmonize and streamline their activities. On the whole, the biobanks in the BBMRI.be network are able to substantially contribute to translational research, as a primary facilitator guaranteeing high quality standards and reproducibility

Mesenchymal stem cells in arthritis

Reumatoide artritis (RA) is een chronische inflammatoire systeemaandoening die zich vooral in het synovium manifesteert. In het proces van synovitis zijn zowel immunologische als inflammatoire processen betrokken, maar daarnaast spelen ook de synoviale fibroblasten een rol en draagt deze stromale celpopulatie bij tot het aantrekken, beïnvloeden en vasthouden van ontstekingscellen. Immunohistochemie (IHC) en celcultuur van synoviale fibroblasten zijn nuttig in het bestuderen van de pathofysiologie van deze aandoening en in de evaluatie van therapie. Cellen die uit het synoviaal membraan (SM) worden geïsoleerd en in cultuur gebracht, voldoen bovendien ook aan de minimale definitie van mesenchymale stamcellen (MSCs) in vitro. In vivo vertonen zij bijkomende kenmerken van weefselplasticiteit. Naast dit vermogen tot differentiatie naar mesenchymale celtypes vertonen MSCs ook immuunmodulerende eigenschappen, waarbij zij immuunresponsen in vitro onderdrukken, maar ook tolerantie kunnen uitlokken ten opzichte van zichzelf of andere cellen in een allogene of xenogene omgeving. MSCs afgeleid van beenmerg en vetweefsel zijn zo in staat gebleken om het klinische verloop van collageen-geïnduceerde artritis (CIA), een veelgebruikt model voor RA, te onderdrukken. Dit effect wordt hoofdzakelijk toegeschreven aan inductie van antigen-specifieke regulatoire T-cellen (Tregs).De uit het SM geïsoleerde en in vitro geëxpandeerde celpopulaties met progenitor kenmerken zijn nauwelijks gekarakteriseerd met betrekking tot hun originele context in het weefsel. Gelijkaardig aan andere stamcellen zijn zij mogelijk te situeren in een specifieke niche, waar bepaalde embryonale signaalwegen, zoals bone morphogenetic proteins (BMPs), het gedrag van de MSC populatie sturen. De bestaande evidentie dat BMP signalen geactiveerd zijn in chronische artritis ondersteunt een mogelijke rol voor MSCs in het ziekteproces, waarbij hun normale functie verstoord is. Deze hypothese wordt nog verder gevoed door de evidentie rond het immuunmodulerend potentieel van deze cellen. Om deze vragen te beantwoorden is het nodig om een zo specifiek mogelijke isolatie en karakterisatie van synoviale progenitorcellen uit te voeren, gevolgd door de studie van deze populaties in artritis modellen, die zij mogelijkerwijze moduleren. We concentreerden de studie op de synoviale isolaten, gezien expansie in cultuur leidt tot fenotypische veranderingen afhankelijk van de specifieke condities, die daarenboven de verschillen tussen subpopulaties of stalen kunnen maskeren. We optimaliseerden een kort isolatieprotocol voor synoviale biopten van artritis- en controlepatiënten, bestaande uit enzymatische en mechanische componenten. We analyseerden de isolaten door middel van multilabel flow cytometrie. Hierbij konden macrofagen, B- en T-lymphocyten, en stromale populaties zoals endotheelcellen en pericyten onderscheiden worden. De cellen van hematopoietische oorsprong waren talrijker in de biopten van artritispatiënten ten opzichte van controles. Er was een grotere CD34-expressie in spondyloartritis stalen ten opzichte van RA stalen, wat bestaande literatuurgegevens over de synoviale vascularisatie bevestigt, en hetgeen ook in onze stalen werd bevestigd met IHC. Om de methode nog beter te valideren, bestudeerden we de invloed van de digestieprocedure op de detectie van oppervlaktemerkers. Monocyten en B-lymphocyten bleken respectievelijk gevoelig aan de enzymatische en de mechanische component van de digestie. Na digestie werden primaire celculturen gestart vanuit de isolaten. Het fenotype van deze geëxpandeerde cellen werd niet beïnvloed door de celdensiteit of de groei op een collageen type 1-bodem. Deze SM-MSCs vertoonden een zekere mate van CD34-expressie, in tegenstelling tot beenmerg-gederiveerde MSCs. Het sorteren van CD34-negatieve cellen vanuit de isolaten leidde daarbij echter na expansie niet tot CD34-negatieve SM-MSCs. In de isolaten werden ook kleine CD146- en CD271-positieve celpopulaties gedetecteerd. Een CD271-positieve populatie werd gesorteerd maar verloor deze merkerexpressie tijdens expansie, waardoor ook CD271 niet als merker voor stabiele identificatie kon gebruikt worden. De expressie van BMP-receptors werd flow cytometrisch onderzocht in de SM-MSCs waarbij enkel ALK2 detecteerbaar bleek. In de isolaten werd geen van deze receptoren gedetecteerd, hoewel IHC kleuringen ALK2 expressie aantoonden in controlesynovium en nog meer uitgesproken in artritis, wat suggereert dat de receptor aangetast werd door de digestie procedure. De SM-MSCs van controle- en artritispatiënten vertoonden dosisafhankelijke onderdrukking van de T-cel proliferatie in vitro, parallel aan het effect van andere stromale cellen zoals beenmerg-gederiveerde MSCs. Na lokale transfer in de spier van immuuncompetente muizen werd echter een immuunrespons uitgelokt waarbij de cellen na twee dagen wel nog intact leken, maar na drie weken niet meer detecteerbaar waren. Een systemische transfer van zowel SM-MSCs als MSCs uit humaan vetweefsel naar muizen met CIA, werd uitgevoerd drie weken na de initiële immunisatie. De klinische evolutie van het model werd hierdoor echter niet positief beïnvloed, maar eerder was een toename van de symptomen waar te nemen ten opzichte van niet-behandelde muizen. Op het eindpunt vonden we een sterker verhoogd IL-5 in de behandelde muizen. In een volgend experiment werden de behandelingsmodaliteiten uitgebreid naar hogere dosissen MSCs en meervoudige injecties. Het klinisch verloop werd gevolgd tot dag 35 en verliep gelijk voor muizen die MSCs versus PBS toegediend kregen, en evenmin werden pathologische verschillen gezien. Wel noteerden we tussen dag 23 en 35 een tendens tot lagere IL-5 waarden (Th2 respons) in de muizen die met cellen werden behandeld. Gezien deze resultaten in tegenspraak blijven met de specifieke studies, die een gunstig effect van humane MSCs in CIA beschrijven, is verdere studie van deze opstelling noodzakelijk. Hierbij zouden enerzijds in vivo gegevens over Treg inductie in de behandelde muizen moeten verzameld worden, en anderzijds de invloed van synoviale MSCs op T-cellen in vitro opnieuw moeten onderzocht worden met betrekking tot het verwerven van een regulatoir phenotype.nrpages: 2status: publishe

Automated Sample Storage in Biobanking to Enhance Translational Research: The Bumpy Road to Implementation

The low reproducibility of biomarker research is a major holdback for the translation of research results to the bedside. Sample integrity has been identified as a key factor that contributes to improved reproducibility. The key mission of biobanks is to ensure that all activities and materials are managed according to standardized procedures and best practices to ensure and preserve sample integrity. When handling large numbers of biospecimens automation of sample handling and storage is often the method of choice to maintain and improve sample integrity. In December 2013, the centralized Biobank of the University Hospitals and the Catholic University of Leuven (UZ KU Leuven) decided to implement automated systems for sample storage and retrieval, one for storage at -20°C and one for storage at -80°C. Here we describe the extensive process of installation, acceptance, validation, and implementation of these two systems. Overall it took about 4 years to effectively take the systems into production. Multiple issues resulted in the delayed implementation, with labware change, quality of the initial installation, and misunderstanding of biobank concerns being the most impacting. Significant effort in terms of time and resources from both the automated store supplier as well as the biobank itself was needed to achieve a successful implementation. Within 15 months of actual integration in the biobank workflow, over 63 k samples were placed into the systems. Actual hands-on sample handling and retrieval times were substantially reduced, although this implied the shift of dedicated personnel time from the researchers' laboratories to the biobank. With the successful implementation of automated frozen sample storage systems, the centralized UZ KU Leuven Biobank is now also able to efficiently support large-scale translational research.status: publishe

Biobank Quality Management in the BBMRI.be Network

From as early as 2005, different guidelines and quality standards covering biobank activities and sample handling methods have been developed to improve and guarantee the reproducibility of biomarker research. Ten years on, the BBMRI.be Quality working group wanted to gauge the current situation of these aspects in the biobanks of the BBMRI.be network. To this end, two online surveys were launched (fall 2017 and fall 2018) to the biobank quality managers in the BBMRI.be network to determine the status and setup of their current quality management system (QMS) and how their QMS and related practices have evolved over a 14 month time period. All biobanks addressed by the two surveys provided a complete response (12 and 13, respectively). A QMS was implemented in 85% of biobanks, with 4 standards emerging as primary basis. Supplementary guidelines were used, with a strong preference for the ISBER best practices for biobanks. The Standard Preanalytical Code-an indicator of the preanalytical lifecycle of a biospecimen impacting the downstream analysis results-was already implemented in 50% of the biobanks while the other half intends future implementation. To assess and maintain the quality of their QMS, 62% of biobanks used self-assessment tools and 71% participated in proficiency testing schemes. The majority of biobanks had implemented procedures for general and biobank specific activities. However, policies regarding the business and sustainability aspect of biobank were only implemented in a limited number of biobanks. A clear desire for a peer-review audit was expressed by 69% of biobanks, with over half of them intending to implement the recently published biobank standard ISO20387. Overall, the biobanks of the BBMRI.be network have actively implemented a solid quality approach in their practices. The implementation of ISO 20387 may bring further professionalization of activities. Based on the needs expressed in this survey, the Quality working group will be setting up an audit program for the BBMRI.be biobanks, to enhance, harmonize and streamline their activities. On the whole, the biobanks in the BBMRI.be network are able to substantially contribute to translational research, as a primary facilitator guaranteeing high quality standards and reproducibility.status: publishe

CD248 and its cytoplasmic domain: a therapeutic target for Arthritis

Objective. CD248 is a transmembrane glycoprotein expressed on the surface of activated perivascular and fibroblast-like cells. This study was undertaken to explore the function of CD248 and its cytoplasmic domain in arthritis. Methods. Synovial tissue biopsy samples from healthy controls, from patients with psoriatic arthritis (PsA), and from patients with rheumatoid arthritis (RA) were stained for CD248. Transgenic mice that were CD248-deficient (CD248-knockout [CD248(KO/KO)]) or mice with CD248 lacking the cytoplasmic domain (CD248(CyD/CyD)) were generated. Collagen antibody-induced arthritis (CAIA) was induced in these mice and in corresponding wild-type (WT) mice as controls. Clinical signs and histologic features of arthritis were evaluated. Cytokine levels were determined by enzyme-linked immunosorbent assay, and the number of infiltrating inflammatory cells was quantified by immuno-histochemistry. In vitro studies were performed with fibroblasts from CD248-transgenic mouse embryos to explain the observed effects on inflammation. Results. Immunostaining of synovium from patients with PsA and patients with RA and that from mice after the induction of CAIA revealed strong CD248 expression in perivascular and fibroblast-like stromal cells. CD248(KO/KO) and CD248(CyD/CyD) mice had less severe arthritis, with lower plasma levels of proinflammatory cytokines, as compared with WT controls. Moreover, the joints of these mice had less synovial hyperplasia, reduced accumulation of inflammatory cells, and less articular cartilage and bone damage. Tumor necrosis factor alpha-induced monocyte adhesion to CD248(CyD/CyD) fibroblasts was impaired. CD248(CyD/CyD) fibroblasts exhibited reduced expression of hypoxia-inducible factor 1 alpha, placental growth factor, vascular endothelial growth factor, and matrix metalloproteinase 9 activity in response to transforming growth factor beta. Conclusion. CD248 contributes to synovial hyperplasia and leukocyte accumulation in inflammatory arthritis, the effects of which are mediated partly via its cytoplasmic domain. CD248 is therefore a potential new target in the treatment of arthritis

Inflammatory Bowel Disease (IBD)-A Textbook Case for Multi-Centric Banking of Human Biological Materials

Inflammatory bowel disease (IBD) is a chronic relapsing inflammatory condition affecting mainly the gastro-intestinal tract with two main entities: Crohn's disease (CD) and ulcerative colitis (UC). Although the exact mechanisms underlying the initial development of IBD are not fully understood, it is believed that an abnormal immune response is elicited against the intestinal microbiota in genetically predisposed individuals. Crucial elements of the etiopathogenesis have been elucidated by research using human biological materials. The estimated prevalence of IBD is 0.5% in the Western world. Although incidence rates are increasing, both conditions are not "common" in general terms mandating a multicentric approach. Biological material from numerous Belgian patients have been collected over time in a number of university hospitals in Belgium (UH Ghent: 800 CD patients, 350 UC patients, 600 normal controls; UH Leuven: 2,600 CD patients, 1,380 UC patients, 98 IC/IBDU patients, 6,260 normal controls). Within the setting of the Flemish Center Medical Innovation (CMI) initiative and later on the Flemish biobank network a prospective study was set-up across three Belgian IBD centers (University Hospitals Brussels, Ghent, and Leuven). Human biological materials and data have been collected prospectively from newly diagnosed CD and UC patients. The analyses hereof have generated new insights which have been published in the most renowned journals. The approach of well-thought off, multi-centric, structured, and systematic biobanking has proven to be a success-story and thus a textbook case for multi-centric banking of human biological materials. This story is being told in this article.status: publishe

An EULAR study group pilot study on reliability of simple capillaroscopic definitions to describe capillary morphology in rheumatic diseases

Objective. To propose simple capillaroscopic definitions for interpretation of capillaroscopic morphologies and to assess inter-rater reliability. Methods. The simple definitions proposed were: normal-hairpin, tortuous or crossing; abnormal-not hairpin, not tortuous and not crossing; not evaluable-whenever rater undecided between normal and abnormal. Based upon an aimed kappa of 0.80 and default prevalences of normal (0.4), abnormal (0.4) and not evaluable (0.2) capillaries, 90 single capillaries were presented to three groups of raters: experienced independent raters, n = 5; attendees of the sixth EULAR capillaroscopy course, n = 34; novices after a 1-h course, n = 11. Inter-rater agreement was assessed by calculation of proportion of agreement and by kappa coefficients. Results. Mean kappa based on 90 capillaries was 0.47 (95% CI: 0.39, 0.54) for expert raters, 0.40 (95% CI: 0.36, 0.44) for attendees and 0.46 (95% CI: 0.41, 0.52) for novices, with overall agreements of 67% (95% CI: 63, 71), 63% (95% CI: 60, 65) and 67% (95% CI: 63, 70), respectively. Comparing only normal vs the combined groups of abnormal and not evaluable capillaries did increase the kappa: 0.51 (95% CI: 0.37, 0.65), 0.53 (95% CI: 0.49, 0.58) and 0.55 (95% CI: 0.49, 0.62). On the condition that the capillaries were classifiable, the mean kappa was 0.62 (95% CI: 0.50, 0.74) for expert raters (n = 65), 0.76 (95% CI: 0.69, 0.83) for attendees (n = 20) and 0.81 (95% CI: 0.74, 0.89) for novices (n = 44). Conclusion. This multicentre, international study showed moderate reliability of simple capillaroscopic definitions for describing morphology of capillaries by rheumatologists with varying levels of expertise. Novices were capable of distinguishing normal from abnormal capillaries by means of a 1-h training session. In future studies, the class not evaluable may be obsolete. © The Author 2015