Hyperactive STAT5 hijacks T cell receptor signaling and drives immature T cell acute lymphoblastic leukemia
Tobias Suske, … , Marco Herling, Richard Moriggl
Tobias Suske, … , Marco Herling, Richard Moriggl
Published April 15, 2024
Citation Information: J Clin Invest. 2024;134(8):e168536. https://doi.org/10.1172/JCI168536.
View: Text | PDF
Research Article Genetics Hematology

Hyperactive STAT5 hijacks T cell receptor signaling and drives immature T cell acute lymphoblastic leukemia

Abstract

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive immature T cell cancer. Mutations in IL7R have been analyzed genetically, but downstream effector functions such as STAT5A and STAT5B hyperactivation are poorly understood. Here, we studied the most frequent and clinically challenging STAT5BN642H driver in T cell development and immature T cell cancer onset and compared it with STAT5A hyperactive variants in transgenic mice. Enhanced STAT5 activity caused disrupted T cell development and promoted an early T cell progenitor–ALL phenotype, with upregulation of genes involved in T cell receptor (TCR) signaling, even in absence of surface TCR. Importantly, TCR pathway genes were overexpressed in human T-ALL and mature T cell cancers and activation of TCR pathway kinases was STAT5 dependent. We confirmed STAT5 binding to these genes using ChIP-Seq analysis in human T-ALL cells, which were sensitive to pharmacologic inhibition by dual STAT3/5 degraders or ZAP70 tyrosine kinase blockers in vitro and in vivo. We provide genetic and biochemical proof that STAT5A and STAT5B hyperactivation can initiate T-ALL through TCR pathway hijacking and suggest similar mechanisms for other T cell cancers. Thus, STAT5 or TCR component blockade are targeted therapy options, particularly in patients with chemoresistant clones carrying STAT5BN642H.

Authors

Tobias Suske, Helena Sorger, Gabriele Manhart, Frank Ruge, Nicole Prutsch, Mark W. Zimmerman, Thomas Eder, Diaaeldin I. Abdallah, Barbara Maurer, Christina Wagner, Susann Schönefeldt, Katrin Spirk, Alexander Pichler, Tea Pemovska, Carmen Schweicker, Daniel Pölöske, Emina Hubanic, Dennis Jungherz, Tony Andreas Müller, Myint Myat Khine Aung, Anna Orlova, Ha Thi Thanh Pham, Kerstin Zimmel, Thomas Krausgruber, Christoph Bock, Mathias Müller, Maik Dahlhoff, Auke Boersma, Thomas Rülicke, Roman Fleck, Elvin Dominic de Araujo, Patrick Thomas Gunning, Tero Aittokallio, Satu Mustjoki, Takaomi Sanda, Sylvia Hartmann, Florian Grebien, Gregor Hoermann, Torsten Haferlach, Philipp Bernhard Staber, Heidi Anne Neubauer, Alfred Thomas Look, Marco Herling, Richard Moriggl

×

Figure 6

T-ALL cells with elevated TCR pathway gene expression and STAT5 activation respond to ZAP70 and STAT5 inhibition.

Options: View larger image (or click on image) Download as PowerPoint
T-ALL cells with elevated TCR pathway gene expression and STAT5 activati...
(A) Growth of T-ALL cell lines upon STAT5B or ZAP70 CRISPR/Cas9 knockout via GFP tracing, normalized to day 3 after infection, and AAVS1 and RPL17 as control genes. (B) Western blot analysis for pY-STAT5/-STAT3 and t-STAT5/-STAT3 in KOPT-K1 cells treated 24 hours with JPX-0750 at indicated concentrations or vehicle (Veh), with actin as loading control. (C) Dose-response curve for T-ALL cell lines or human nonleukemic CD3 cells treated with JPX-0750. Three or more independent experiments in technical triplicates were performed. (D) Dose-response synergy analysis of the indicated 2-drug combination in T-ALL cells after 72 hours treatment. The most synergistic area (MSA) is highlighted (white rectangle) with the respective MSA score. One representative plot of 2 independent experiments is shown. (E) Percentage survival of nonleukemic control and T-ALL cells from primary patient samples treated for 24 hours with JPX-0750 at 100, 1,000, or 10,000 nM. Mean values of respective concentration versus DMSO control in 2 technical replicates are shown. Relapsed patients are marked in red. (F) Confocal microscopy of LOUCY, JURKAT, and KOPT-K1 T-ALL cells stained for STAT5B (green) and DAPI (blue), and the number of peaks found in STAT5B ChIP-Seq of respective cell lines. (G) Fractions of hCD45+ KOPT-K1 cells in spleen, liver, and blood after transplantation into NSG mice and treatment with JPX-0750 (n = 8) alone or in combination with fostamatinib (n = 6) or vehicle (n = 7) for 21 days. (H) Representative immunohistochemical staining for CD3 in the spleens and lungs of these mice. Original magnification: ×10; scale bar: 400 μm. In G, significant differences are indicated as *P < 0.05, **P < 0.01 by 1-way ANOVA with Tukey’s multiple comparison test. In A, C, and G, data are shown as the mean ± SEM.