Abstract
Lack of suitable in vitro culture conditions for primary acute lymphoblastic leukaemia (ALL) cells severely impairs their experimental accessibility and the testing of new drugs on cell material reflecting clonal heterogeneity in patients. We show that Nestin-positive human mesenchymal stem cells (MSCs) support expansion of a range of biologically and clinically distinct patient-derived ALL samples. Adherent ALL cells showed an increased accumulation in the S phase of the cell cycle and diminished apoptosis when compared with cells in the suspension fraction. Moreover, surface expression of adhesion molecules CD34, CDH2 and CD10 increased several fold. Approximately 20% of the ALL cells were in G0 phase of the cell cycle, suggesting that MSCs may support quiescent ALL cells. Cellular barcoding demonstrated long-term preservation of clonal abundance. Expansion of ALL cells for >3 months compromised neither feeder dependence nor cancer initiating ability as judged by their engraftment potential in immunocompromised mice. Finally, we demonstrate the suitability of this co-culture approach for the investigation of drug combinations with luciferase-expressing primograft ALL cells. Taken together, we have developed a preclinical platform with patient-derived material that will facilitate the development of clinically effective combination therapies for ALL.
Highlights
Robust preclinical models for childhood acute lymphoblastic leukaemia (ALL) are essential for dissecting mechanisms that drive malignant growth and survival and to test and develop novel targeted therapies that may improve current therapies with regard to efficacy and toxicity
mesenchymal stem cells (MSCs) support short-term proliferation of a range of clinically distinct patient-derived ALL cells To compare the impact of stromal cell lines and primary human MSCs on B-ALL propagation, we examined ALL cell proliferation on either the murine stromal cell line M2-10B4, human TERT-immortalised MSC or primary human MSCs obtained from hip surgery
Primary MSCs were characterised as NES+ CD73+ Cadherin 2 (CDH2)+ CD90++ TEK+; absence of haematopoietic cells was confirmed by lack of CD45 and CD19 surface expression (Supplementary Figure S3)
Summary
Robust preclinical models for childhood acute lymphoblastic leukaemia (ALL) are essential for dissecting mechanisms that drive malignant growth and survival and to test and develop novel targeted therapies that may improve current therapies with regard to efficacy and toxicity. Cell line models have been widely used in functional in vitro studies and preclinical drug screens.[1,2,3,4,5,6] cell lines do retain the original driver mutations, they do not represent the molecular complexity of the disease at presentation. The combination of low complexity and reduced dependence on cell-extrinsic signalling can affect the translation of cell line data to the clinical situation, for example, in relation to clinically relevant mechanisms of drug resistance; 7–9 affecting the ability of cell line models to reflect the original disease. ALL blasts are highly dependent on their in vivo environment and rapidly undergo apoptosis ex vivo.[10,11,12] Shortterm in vitro assays have been developed for testing drug sensitivity;[13] their use has not been widely implemented because of the rapid decline of ALL cells in these assays, even without exposure to any anti-leukaemic compounds
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