Investigating the bone marrow microenvironment in MGUS and multiple myeloma – evidence for early and irreversible transformation Daniel A Tennant Institute of Metabolism and Systems Research University of Birmingham

Investigating a changing network through its metabolic footprint Read-out of: Cell number Cell phenotype Microenvironment Cell-cell communication Haas et al. TiBS (2016)

Metabolic transformation of the bone marrow is an early event in the aetiology of MM Changes in the bone marrow phenotype are early Ludwig et al. Blood Cancer J (2015)

Multiple complex changes across the bone marrow metabolic network Ludwig et al. Blood Cancer J (2015)

This metabolic change can be observed in the peripheral blood Observable change is significant, but small Ludwig et al. Blood Cancer J (2015)

Enhanced urea – signs of early altered bone marrow function Blair et al. Pract Neurol (2014)

BMMSCs are a critical stromal cell type supporting plasma cells Hideshima and Anderson. Nature Rev Cancer (2002)

Gene expression signature of BMMSCs changes with disease onset Ctrl MGUS MM Primary BMMSCs cultured for <5 passages McNee et al. (2017) Leukemia

PADI2 expression is high in MGUS and MM bone marrow mesenchymal stem cells Previously-described PADI2 target Urea transporter McNee et al. (2017) Leukemia

Peptidyl arginine deiminases Family of 5 enzymes that deiminate peptidyl arginyl residues to form citrulline Jang et al. J Bacteriol Virol (2013)

Action of the PADI enzymes

A significant increase in PADI2 expression is observed in BMMSCs from both MGUS and MM McNee et al. (2017) Leukemia

PADI2 expression correlates with IL-6 expression in in vitro and in vivo McNee et al. (2017) Leukemia

IL-6 expression is downstream of PADI2 catalytic activity McNee et al. (2017) Leukemia

Citrullination of arginine 26 of histone H3 is increased in primary BMMSCs McNee et al. (2017) Leukemia

PADI2 activity results in citrullination of the IL-6 promoter region McNee et al. (2017) Leukemia

PADI2 modulates expression of multiple chemokines with a role in myeloma pathogenesis McNee et al. (2017) Leukemia

PADI2 activity modulates therapy responsiveness of co-cultured myeloma cells McNee et al. (2017) Leukemia

Early changes in BMMSC phenotype alter myeloma cell behaviour How does PADI2 become upregulated in early disease? McNee et al. (2017) Leukemia

Acknowledgements University of Birmingham Clare Davies Federica Cuozzo Katherine Eales Cristina Escribano Kate Hollinshead Gavin McNee Haydn Munford Katarina Kluckova Alpesh Thakker National NMR facility Christian Ludwig Clare Davies Paul Moss Sarah Essex David Bartlett Guy Pratt Royal Wolverhampton Hospital NHS Trust Supratik Basu Angelique Barkhuizen