RUNX2 regulates leukemic cell metabolism and chemotaxis in high-risk T cell acute lymphoblastic leukemia
Filip Matthijssens, … , Pieter Van Vlierberghe, Ksenia Matlawska-Wasowska
Filip Matthijssens, … , Pieter Van Vlierberghe, Ksenia Matlawska-Wasowska
Published February 8, 2021
Citation Information: J Clin Invest. 2021;131(6):e141566. https://doi.org/10.1172/JCI141566.
View: Text | PDF
Research Article Oncology

RUNX2 regulates leukemic cell metabolism and chemotaxis in high-risk T cell acute lymphoblastic leukemia

Abstract

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy with inferior outcome compared with that of B cell ALL. Here, we show that Runt-related transcription factor 2 (RUNX2) was upregulated in high-risk T-ALL with KMT2A rearrangements (KMT2A-R) or an immature immunophenotype. In KMT2A-R cells, we identified RUNX2 as a direct target of the KMT2A chimeras, where it reciprocally bound the KMT2A promoter, establishing a regulatory feed-forward mechanism. Notably, RUNX2 was required for survival of immature and KMT2A-R T-ALL cells in vitro and in vivo. We report direct transcriptional regulation of CXCR4 signaling by RUNX2, thereby promoting chemotaxis, adhesion, and homing to medullary and extramedullary sites. RUNX2 enabled these energy-demanding processes by increasing metabolic activity in T-ALL cells through positive regulation of both glycolysis and oxidative phosphorylation. Concurrently, RUNX2 upregulation increased mitochondrial dynamics and biogenesis in T-ALL cells. Finally, as a proof of concept, we demonstrate that immature and KMT2A-R T-ALL cells were vulnerable to pharmacological targeting of the interaction between RUNX2 and its cofactor CBFβ. In conclusion, we show that RUNX2 acts as a dependency factor in high-risk subtypes of human T-ALL through concomitant regulation of tumor metabolism and leukemic cell migration.

Authors

Filip Matthijssens, Nitesh D. Sharma, Monique Nysus, Christian K. Nickl, Huining Kang, Dominique R. Perez, Beatrice Lintermans, Wouter Van Loocke, Juliette Roels, Sofie Peirs, Lisa Demoen, Tim Pieters, Lindy Reunes, Tim Lammens, Barbara De Moerloose, Filip Van Nieuwerburgh, Dieter L. Deforce, Laurence C. Cheung, Rishi S. Kotecha, Martijn D.P. Risseeuw, Serge Van Calenbergh, Takeshi Takarada, Yukio Yoneda, Frederik W. van Delft, Richard B. Lock, Seth D. Merkley, Alexandre Chigaev, Larry A. Sklar, Charles G. Mullighan, Mignon L. Loh, Stuart S. Winter, Stephen P. Hunger, Steven Goossens, Eliseo F. Castillo, Wojciech Ornatowski, Pieter Van Vlierberghe, Ksenia Matlawska-Wasowska

×

Figure 3

RUNX2 regulates T-ALL survival.

Options: View larger image (or click on image) Download as PowerPoint
RUNX2 regulates T-ALL survival. (A) Cell growth of LOUCY and KARPAS-45 c...
(A) Cell growth of LOUCY and KARPAS-45 cells transduced with RUNX2 shRNAs (shRUNX2 B, shRUNX2 C) and scrambled control (shNC). Data are shown as mean ± SD for 1 of 3 independent experiments performed in triplicate. (B) Flow cytometry cell-cycle analysis using propidium iodide staining (48 hours). (C) Apoptotic cell death in T-ALL cells stained with annexin V/7-AAD (96 hours). (B and C) Data are representative of 1 of 3 independent experiments. (D) Human LOUCY and KARPAS-45 cells, murine MOHITO cells expressing control plasmid (CON), KMT2A-R (KMT2A-MLLT1, KMT2A-MLLT4), and primary T-ALL cells (PASSPP, 18-169) were transduced with RUNX2 shRNAs and scrambled control. Western blot analysis of the indicated proteins. (E) RUNX2 and H3K27ac ChIP-Seq at the CTNNB1 (β-catenin) and BIRC5 (survivin) loci in KARPAS-45. Forced expression of (F) active β-catenin and (G) survivin in LOUCY with or without shRNA-mediated RUNX2 depletion. (H and I) Annexin V/7-ADD staining in LOUCY (96 hours). Data are shown as mean ± SD for 3 independent experiments. AnV, annexin V. (J) Kaplan-Meier survival curve analyses of NSG mice (n = 10/group) transplanted with 106 LOUCY cells expressing shRUNX2 B, shRUNX2 C, or scrambled control (median log-rank Mantel-Cox test). (K) Flow cytometric quantification of human CD45+ cells in mice (n = 4/group) euthanized 41 days after inoculation with 106 transduced LOUCY cells. (L) The levels of GFP+ cells were determined in blood (day 30), BM, and spleen (day 100) in animals injected with PDX samples harboring KMT2A-MLLT-1. (A) Repeated measure ANOVA with Tukey’s multiple comparisons test; (H, I, K, and L) 1-way ANOVA with Tukey’s multiple comparison test. ***P < 0.0005; ****P < 0.0001. (D, F, and G) Representative blots from at least 2 separate experiments.