PB2092 IMPLEMENTATION AND VALIDATION OF A COLLABORATIVE PLATFORM FOR LONGITUDINAL BIOBANKING OF VIABLE CELLS AND PLASMA IN MYELOID NEOPLASMS

Abstract

Background:Myeloid malignancies are heterogeneous disorders affecting proliferation and differentiation of hematopoietic stem cells (HSCs). They comprise myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPN) as well as overlap MDS/MPN syndromes and are characterized by a propensity to evolve towards secondary acute myeloid leukemias (AML). The underlying genetic composition is very heterogeneous and involves sequential accumulation of mutations in genes of epigenetic regulation, transcriptional control, splicing, signal transduction as well as cell cycling. Due to the complex genetic interactions and the unpredictable clonal evolution, longitudinal collection of biologic material is mandatory to forward our understanding of the pathogenesis in myeloid malignancies. The implementation of a biobanking platform is, therefore, important to foster translational research in the current post‐genomic era of “precision medicine”.Aims:We aimed to establish a standardized platform for longitudinal collection and storage of viable cells as well as plasma from peripheral blood (PB) and bone marrow (BM) for patients treated for myeloid malignancies at our institution.Methods10‐20 mL of PB/BM were collected and processed with isolation of mononuclear cells (MNCs) using density‐gradient centrifugation. 1 mL PB/BM was separately centrifuged and 500ul plasma stored at ‐80 °C. Manual and automated measurement of cell‐counts as well as viability was performed. MNCs were stored at a maximum concentration of 10 × 10e6 cells in 500 μL freezing‐medium (RPMI, 10% DMSO, 20% FCS) using 0.7 mL 2D‐barcoded cryotubes (FluidX®). Samples were immediately frozen at <‐150 °C with an automated freezing work bench using a predefined temperature curve and centrally stored in a fully‐automated biobank (ASKION C‐line® system). Relevant health related data was simultaneously collected on a SecuTrial web‐based database.Results:Between March 2017 and January 2019, 122 patient samples were included in our biobank. Samples originated from BM (MNCs n = 108; plasma n = 23), PB (MNCs n = 57) and leukapheresis (n = 9). Samples were mainly collected at diagnosis and conditions comprised AML (n = 16), MDS/MPN (n = 13), MDS (n = 14), hypoplastic bone‐marrow conditions (hypoplastic MDS/aplastic anaemia: n= 5), idiopathic cytopenia of unknown significance (ICUS: n = 26) and others (n = 26) (Table 1). The median volume of BM collected was 10 mL. BM cell‐counts varied strongly with the highest values in MDS/MPN patients. MDS and ICUS demonstrated the lowest cell‐count and also viability compared to other conditions. Hypoplastic bone marrow conditions showed low cell‐count with paradoxically high viabilities. 19 frozen MNCs samples from patients with different conditions were thawed and we observed cell viabilities ranging from 51‐92%. Due to small sample size, no correlation of post‐thawing viabilities with clinical conditions could be made. We observed clumping of cells after thawing, which was associated with delay in processing (< 25% clumping with immediate processing; 50% after 24 h; >75% after more than 24 h).Summary/Conclusion:The implementation of a shared biobank platform for longitudinal collection of viable cells poses numerous challenges at personnel, infrastructural, logistic, technical and financial levels. Such a platform is feasible but collection procedures must be highly standardized at all levels to minimize heterogeneity of sample quality. Our preliminary experience provides an essential basis for further improvement of our platform and will be instrumental to foster collaborative translational research in myeloid neoplasms.image