Community Research and Development Information Service - CORDIS

ERC

BioBlood Report Summary

Project ID: 340719
Funded under: FP7-IDEAS-ERC
Country: United Kingdom

Mid-Term Report Summary - BIOBLOOD (Development of a Bio-Inspired Blood Factory for Personalised Healthcare)

BioBlood is an ambitious interdisciplinary project that aims to deliver a “step-change” in precision cellular and drug therapies for patients with blood disorders, specifically those requiring red blood cell (RBC) transfusions and treatments for acute myeloid leukaemia (AML) and chronic lymphocytic leukaemia (CLL). To this end, we have thus far manufactured a 3D hollow fibre perfusion bioreactor (HFB) for the cultivation of human cord blood (for RBC production) and primary human bone marrow and peripheral blood samples from patients with AML and CLL (for evaluation of treatment). One of the main objectives of the project is to manufacture RBCs for eventual transfusion into patients; the HFB construct has therefore been optimised towards a system which could eventually be used for clinical purposes (GMP-compliant) by (1) modification of the 3D scaffold by the attachment of RGD tripeptide to enhance cell adhesion and growth, replacing collagen coating, (2) improvement of the alumina hollow fibres to ensure stability during the HFB manufacturing process and a pore size combination of the outer and inner surfaces which would improve selective RBC filtration and harvest and, (3) HFB miniaturization in order to accommodate robust growth of smaller cord blood and patient samples. For the HFB bioprocess (in vitro), we have cultured human umbilical cord blood with serum-free media appropriate for clinical translation, supplemented with near-physiologic cytokine concentrations in order to produce normal RBCs, harvested and filtered out of the HFB; in situ analyses revealed that the 3D HFB microenvironment supported maturing RBC niches, including stromal populations and supportive growth factors, which changed in spatial conformity over time of the long-term culture. A novel mathematical tool (in silico) was developed to quantitate these cell associations over space and time within the HFB in order to provide a means of targeting improvements in the platform for the next project phases. We have also optimised the 3D culture techniques of cord blood towards RBCs to incorporate low oxygen (hypoxia) to enhance physiologic erythropoiesis. Imitating what would normally occur in the bone marrow, a combination schedule of early stage hypoxia with late stage normal oxygen exposure used in standard cultures (normoxia) extended erythroid maturation, sustained culture self-renewal and created an in vitro 3D microenvironment with supportive cellular and cytokine niches. This hypoxia-normoxia schedule will now be incorporated into the perfusion HFB RBC production platform. The second main objective of BioBlood is to create a 3D culture (in vitro) and mathematical model expression (in silico) of human AML and CLL throughout treatment. Thus far, we have acquired ethics and R&D approvals to acquire anonymised patient samples and datasets for use in the project. Using these patient samples, long-term 3D static cultures of AML and CLL have been achieved in cytokine-free conditions and we have determined optimum seeding density and culture conditions for both. In order to investigate the role of the microenvironment and parameters for the bioprocess, a metabolomics protocol has been established for primary and cultured AML samples, results for which have highlighted the importance of glycolysis in AML. These leukaemia cultures will now be moved into the smaller perfusion HFB, “mini-reactors”, to assess leukaemia progression, response to treatment, and for use in the in silico modelling of precision chemotherapy for AML and CLL. In order to achieve precision, we have developed a Population Balance Model (PBM) which incorporates experimentally-derived cyclin parameters within the mathematical description of the cell cycle to describe the heterogeneity and kinetics of leukaemia cells, critical to describing response and resistance to treatment. Using PBM and experimental validation with leukaemia cell lines, we have been able to predict leukaemic clonal kinetics over time and derive clonal origins. An in silico model of CLL disease progression has also been created and, using global sensitivity analysis, critical parameters in CLL patients were found to be proliferation in nodal centres (bone marrow and lymph nodes) and migration rates between them, an area in CLL treatment which has not previously been addressed. We are currently employing historical patient datasets in AML and CLL in order to test applicability of the in silico PBM model in the treatment setting, integrating pharmacokinetics-pharmacodynamics with growth kinetics and cell cycle parameters of leukaemia. In the next phases of BioBlood, this model will be used in tandem with the in vitro HFB AML and CLL models before, during and after treatment.

Contact

Shaun Power, (Research Services Manager)
Tel.: +44 207 594 8773
Fax: +44 207 594 8609
E-mail
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