DYRK1A regulates B cell acute lymphoblastic leukemia through phosphorylation of FOXO1 and STAT3
Rahul S. Bhansali, Malini Rammohan, Paul Lee, Anouchka P. Laurent, Qiang Wen, Praveen Suraneni, Bon Ham Yip, Yi-Chien Tsai, Silvia Jenni, Beat Bornhauser, Aurélie Siret, Corinne Fruit, Alexandra Pacheco-Benichou, Ethan Harris, Thierry Besson, Benjamin J. Thompson, Young Ah Goo, Nobuko Hijiya, Maria Vilenchik, Shai Izraeli, Jean-Pierre Bourquin, Sébastien Malinge, John D. Crispino
Rahul S. Bhansali, Malini Rammohan, Paul Lee, Anouchka P. Laurent, Qiang Wen, Praveen Suraneni, Bon Ham Yip, Yi-Chien Tsai, Silvia Jenni, Beat Bornhauser, Aurélie Siret, Corinne Fruit, Alexandra Pacheco-Benichou, Ethan Harris, Thierry Besson, Benjamin J. Thompson, Young Ah Goo, Nobuko Hijiya, Maria Vilenchik, Shai Izraeli, Jean-Pierre Bourquin, Sébastien Malinge, John D. Crispino
View: Text | PDF
Research Article Oncology

DYRK1A regulates B cell acute lymphoblastic leukemia through phosphorylation of FOXO1 and STAT3

Abstract

DYRK1A is a serine/threonine kinase encoded on human chromosome 21 (HSA21) that has been implicated in several pathologies of Down syndrome (DS), including cognitive deficits and Alzheimer’s disease. Although children with DS are predisposed to developing leukemia, especially B cell acute lymphoblastic leukemia (B-ALL), the HSA21 genes that contribute to malignancies remain largely undefined. Here, we report that DYRK1A is overexpressed and required for B-ALL. Genetic and pharmacologic inhibition of DYRK1A decreased leukemic cell expansion and suppressed B-ALL development in vitro and in vivo. Furthermore, we found that FOXO1 and STAT3, transcription factors that are indispensable for B cell development, are critical substrates of DYRK1A. Loss of DYRK1A-mediated FOXO1 and STAT3 signaling disrupted DNA damage and ROS regulation, respectively, leading to preferential cell death in leukemic B cells. Thus, we reveal a DYRK1A/FOXO1/STAT3 axis that facilitates the development and maintenance of B-ALL.

Authors

Rahul S. Bhansali, Malini Rammohan, Paul Lee, Anouchka P. Laurent, Qiang Wen, Praveen Suraneni, Bon Ham Yip, Yi-Chien Tsai, Silvia Jenni, Beat Bornhauser, Aurélie Siret, Corinne Fruit, Alexandra Pacheco-Benichou, Ethan Harris, Thierry Besson, Benjamin J. Thompson, Young Ah Goo, Nobuko Hijiya, Maria Vilenchik, Shai Izraeli, Jean-Pierre Bourquin, Sébastien Malinge, John D. Crispino

×

Figure 7

DYRK1A-mediated phosphorylation of STAT3 regulates mitochondrial ROS in B-ALL.

Options: View larger image (or click on image) Download as PowerPoint
DYRK1A-mediated phosphorylation of STAT3 regulates mitochondrial ROS in ...
(A) Western blot results of an in vitro kinase assay with GST-DYRK1A, FLAG-STAT3, and ATP-γ-S. Alkylated reaction products were analyzed for thiophosphate esters. (B) Western blot showing p-STAT3 (Ser727, Tyr705) and total STAT3 protein in MUTZ-5 cells after 4 hours of treatment with EHT 1610. Densitometric values were normalized to actin. (C) Western blot showing p-STAT3 (Ser727, Tyr705), total STAT3, and DYRK1A in MUTZ-5 cells with or without CRISPR/Cas9-mediated knockout of DYRK1A (i, ii, and iii indicate separate blots from the same cell extracts). (D) IC50 values of MUTZ-5 cells transduced with MSCV-puro (empty, STAT3WT, STAT3S727A, STAT3S727D, or STAT3S727E) after treatment with 3 μM EHT 1610. Data indicate the mean ± SD. (E) Quantification of flow cytometric analysis of apoptosis using annexin V staining in MUTZ-5 cells transduced with MSCV-puro (empty, STAT3WT, STAT3S727A, STAT3S727D, or STAT3S727E) after treatment with 3 μM EHT 1610. Data indicate the mean ± SD. (F) Mitochondrial ROS luminescence quantification as RLU (relative light units) after treatment with 3 μM EHT 1610 using the Mitochondrial Superoxide Assay Kit in MUTZ-5 cells transduced with MSCV-puro (empty, STAT3WT, STAT3S727A, STAT3S727D, or STAT3S727E). (G) Cell numbers after treatment of B-ALL cell lines with C188-9, quantified by luminescence as RLU. Curves were fitted nonlinearly with variable slope (4 parameters). IC50 values are shown. (H) Cell numbers after treatment of human PDX–passaged Ph-like ALL samples with C188-9, quantified by luminescence as RLU. Curves were fitted nonlinearly with variable slope (4 parameters). IC50 values are shown. n = 3 biological replicates (A–H). Significance was determined by ANOVA with post hoc Bonferroni’s correction (D–F). ***P < 0.001 and ****P < 0.0001.