Abstract

Relapse remains a leading cause of treatment failure in acute myeloid leukaemia (AML), making haematopoietic stem cell transplantation (HSCT) the only curative option for many patients. Yet HSCT efficacy is often limited by impaired engraftment, driven by AML-induced remodelling of the bone marrow stem cell niche. Mesenchymal stromal cells (MSCs) are key mediators of niche formation and could, in principle, restore a supportive microenvironment when introduced alongside HSC therapy; but this strategy remains largely untested. A key obstacle to MSC-based therapy is that MSCs acquire a senescent, pro-inflammatory phenotype during standard in vitro expansion. We addressed this by engineering a polymer-laminin presentation system that suppresses senescence and preserves a proliferative, regenerative MSC phenotype during expansion. Then, to investigate potential cell therapy use, we developed a bioengineered in vitro model as a new approach methodology (NAM) for studying disease-driven niche modification. The system consists of MSC spheroids embedded in a synthetic hydrogel within a transwell platform, allowing controlled co-culture of healthy or AML-derived haematopoietic cells, therapeutic MSCs, and chemotherapeutic agents. Using this platform, we modelled an AML-like niche and showed that MSCs expanded via the polymer-laminin system, when introduced alongside HSCs, significantly improved HSCT engraftment relative to both standard-expanded MSCs and HSCT performed without MSC support. These results establish MSC phenotype maintenance as a critical determinant of therapeutic efficacy, and position this NAM as a platform for pre-clinical screening of niche-targeted therapies in AML.