Introduction and background Dissecting Roles of Dynein Light Chain 1 towards The Novel Therapy for Lymphomas Lingli Li, PhD candidate, Year 3, Department of Medicine/ School of Medicine, Dentistry and Health sciences, University of Melbourne /SVI @lilly9612 Introduction and background B cell lymphomas are blood cancers arise from B cell development defects . Cell circle and apoptosis are critical steps for maintaining cell numbers during normal B cell development. B-lymphocytes (B cells) guard our health and are responsible for humoral immunity by producing antibodies required. Dynein Light Chain 1 Dynein light chain 1 (DYNLL1) is part of the cytoplasmic dynein motor complex Structural Role: DYNLL1 Mark P Dodding and Michael Way The EMBO Journal (2011) DYNLL1 binds to multiple proteins regulates protein expression in cell cycle, apoptosis etc Gene Regulator: Jurado, S, J Biol Chem. 2012 B cells are continuously generated from precursors in the bone marrow, development and differentiation are mainly regulated by BCR (B cell receptor) and Pre-BCR (Pre B cell receptor) signals. More than half of the aggressive types of NHL are incurable with poor three-year survival (30-50%) and severe side effects under current treatments. Target therapy becomes the focus of search for effective treatment of lymphomas.

Important steps for early B cell development B cell Tumorigenesis Ulf Klein & Riccardo Dalla-Favera. 2008 Important steps for early B cell development Modified from Hardy, R et al, Annul Review Immunology 19, 595-621 (2001) in Heierhorst lab, SVI Conditional Deletion of Dynll1 using Cre Recombination Research Strategy Heierhorst lab data SVI

Results 1: DYNLL1 in early B cell development Reduction of Splenic B cells to 12% B cells in Periphery Blood and in Spleen Decres Cell cycle Profile shows in Pre B cells: Cell division slow down G1 C’ B cells in blood Ctrl Del B cells B cells in spleen Deletion of Dynll1: Huge reduction of B cells in periphery blood and in spleen Deletion of Dynll1 Decrease of the cycling fraction C’ fraction of Pre B, fewer dividing cells] More cells pause at G1 phase before entering to cell cycling. B Cells Pre B cells Snapshot of B Cell Fractions in Bone Marrow Immature B Decres Apoptosis analysis showed that in Immature B cells: Cell death Up Cell Death Deletion of Dynll1 in Bone marrow Big Loss of Pre B cells( C’ + D ) Huge loss of immature B cells (E) Deletion of Dynll1 Increase of apoptosis rate, immature B cells die fast before leave bone marrow. B Cells References References LeBien, T. W. & Tedder, T. F. B , Blood 112, 1570-1580 (2008). Hardy, R et al, Annul Review Immunology 19, 595-621 (20001) Kuppers, R. Nat Rev Cancer 5, 251-262 (2005). Jurado, S.,et al J Exp Med 209, 1629-1639 (2012). Adams, J. M et al, Nature 318, 533-538 (1985). Jurado, S. et al, J Biol Chem 287, 3156-3164 (2012). Rapali, P et al, FEBS J 278, 2980-2996 (2011). Puthalakath, H et al Mol Cell 3, 287-296 (1999). Bouillet, P et al, Science 286, 1735-1738 (1999). Hobeika, E et al, Proc Natl Acad Sci U S A 103, 13789-13794 (2006).   LeBien, T. W. & Tedder, T. F. B , Blood 112, 1570-1580 (2008). Hardy, R et al, Annul Review Immunology 19, 595-621 (20001) Kuppers, R. Nat Rev Cancer 5, 251-262 (2005). Jurado, S.,et al J Exp Med 209, 1629-1639 (2012). Adams, J. M et al, Nature 318, 533-538 (1985). Jurado, S. et al, J Biol Chem 287, 3156-3164 (2012). Rapali, P et al, FEBS J 278, 2980-2996 (2011). Puthalakath, H et al Mol Cell 3, 287-296 (1999). Bouillet, P et al, Science 286, 1735-1738 (1999). Hobeika, E et al, Proc Natl Acad Sci U S A 103, 13789-13794 (2006).  

Results 2: DYNLL1 in Lymphomagenesis Dramatic reduction of immature B in Pre Tumor Mice Survival Analysis of Leukemic Mice Ctrl Del Double Mut Wong, D et al, Cell Rep. 16;14(6):1488-99 (2016). Deletion of Dynll1 in Myc driven pre tumor mice: Huge reduction of Immature B and circulating B cell numbers Immature B cell fraction in mice Deletion of Dynll1 in tumor mice, protects mice from tumor formation: Significantly increase the life span of tumor mice Significant fewer mice develop tumors Fewer mice develop lymphomas There is significant increase in life span when deleted Dynll1.  

References References Conclusions Dynll1 is essential for early B cell development in the bone marrow: regulates apoptosis processes in immature B cells facilitates cell cycle entry at the Pre B cell stage The significant protection of Dynll1-deleted mice from lymphoma formation indicates that DYNLL1 might be a novel target for therapeutic strategies to treat B cell lymphomas. Acknowledgements: Supervisory and Advisory committee: A/P Jörg Heierhorst, A/Prof Jock Campbell, Dr Ora Obernard, A/ Prof Carl Walkley, Dr Meaghan Wall Bio Resources Centre in St Vincent’s Hospital Funding: APA University of Melbourne Scholarship and Leukaemia foundation PhD Scholarship Host institute: SVI and University of Melbourne References: Adams, J. M et al, Nature 318, 533-538 (1985). Bouillet, P et al, Science 286, 1735-1738 (1999). Dupunt, T et al, Oncotarget. 19;7(3):3520-32 (2016). Hardy, R et al, Annul Review Immunology 19, 595-621 (2001). Hobeika, E et al, Proc Natl Acad Sci U S A 103, 13789-13794 (2006). Jurado, S.,et al J Exp Med 209, 1629-1639 (2012). Jurado, S. et al, J Biol Chem 287, 3156-3164 (2012). Kuppers, R. Nat Rev Cancer 5, 251-262 (2005). LeBien, T. W. & Tedder, T. F. B , Blood 112, 1570-1580 (2008). Mark P Dodding and Michael Way, The EMBO Journal (2011). Rapali, P et al, FEBS J 278, 2980-2996 (2011). Puthalakath, H et al Mol Cell 3, 287-296 (1999). Ulf Klein & Riccardo Dalla-Favera. Nature Reviews Immunology 8, 22-33 (2008). Roberts AW et al, N Engl J Med28;374(4):311-22 (2016). Xiao Q, et al, Int J Biol Sci 12(9):1083-1092 (2016). Wong, D et al, Cell Rep. 16;14(6):1488-99 (2016). All experimental results used in this poster have been generated using Heierhorst lab resources in SVI by the author, some are yet to be published. References References LeBien, T. W. & Tedder, T. F. B , Blood 112, 1570-1580 (2008). Hardy, R et al, Annul Review Immunology 19, 595-621 (20001) Kuppers, R. Nat Rev Cancer 5, 251-262 (2005). Jurado, S.,et al J Exp Med 209, 1629-1639 (2012). Adams, J. M et al, Nature 318, 533-538 (1985). Jurado, S. et al, J Biol Chem 287, 3156-3164 (2012). Rapali, P et al, FEBS J 278, 2980-2996 (2011). Puthalakath, H et al Mol Cell 3, 287-296 (1999). Bouillet, P et al, Science 286, 1735-1738 (1999). Hobeika, E et al, Proc Natl Acad Sci U S A 103, 13789-13794 (2006).   LeBien, T. W. & Tedder, T. F. B , Blood 112, 1570-1580 (2008). Hardy, R et al, Annul Review Immunology 19, 595-621 (20001) Kuppers, R. Nat Rev Cancer 5, 251-262 (2005). Jurado, S.,et al J Exp Med 209, 1629-1639 (2012). Adams, J. M et al, Nature 318, 533-538 (1985). Jurado, S. et al, J Biol Chem 287, 3156-3164 (2012). Rapali, P et al, FEBS J 278, 2980-2996 (2011). Puthalakath, H et al Mol Cell 3, 287-296 (1999). Bouillet, P et al, Science 286, 1735-1738 (1999). Hobeika, E et al, Proc Natl Acad Sci U S A 103, 13789-13794 (2006).