Development of combination therapies with BTK inhibitors and dasatinib to treat CNS-infiltrating E2A-PBX1+/preBCR+ ALL.
Gaia Gentile, Teresa Poggio, Antonella Catalano, Minna Voutilainen, Mari Lahnalampi, Marta Andrade-Martinez, Tobias Ma, Roman Sankowski, Lina Goncharenko, Stefan Tholen, Kyuho Han, David W Morgens, Marco Prinz, Michael Lübbert, Sophia Engel, Tanja Nicole Hartmann, Gunnar Cario, Martin Schrappe, Lennart Lenk, Martin Stanulla, Justus Duyster, Peter Bronsert, Michael C Bassik, Michael L Cleary, Oliver Schilling, Merja Heinäniemi, Jesús Duque-Afonso
Author Information
Gaia Gentile: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Teresa Poggio: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
Antonella Catalano: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Minna Voutilainen: Institute of Biomedicine, School of Medicine, University of Eastern Finland, Kuopio, Finland.
Mari Lahnalampi: Institute of Biomedicine, School of Medicine, University of Eastern Finland, Kuopio, Finland. ORCID
Marta Andrade-Martinez: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
Tobias Ma: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Roman Sankowski: Department of Neuropathology, University of Freiburg Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
Lina Goncharenko: Institute for Molecular Medicine and Cell Research, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Stefan Tholen: Institute of Surgical Pathology, University of Freiburg Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
Kyuho Han: Department of Genetics, Stanford University School of Medicine, Stanford, CA. ORCID
David W Morgens: Department of Genetics, Stanford University School of Medicine, Stanford, CA.
Marco Prinz: Department of Neuropathology, University of Freiburg Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Michael Lübbert: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Sophia Engel: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
Tanja Nicole Hartmann: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
Gunnar Cario: Department of Pediatrics, University Medical Center Schleswig-Holstein, Campus Kiel, Kiel, Germany.
Martin Schrappe: Department of Pediatrics, University Medical Center Schleswig-Holstein, Campus Kiel, Kiel, Germany. ORCID
Lennart Lenk: Department of Pediatrics, University Medical Center Schleswig-Holstein, Campus Kiel, Kiel, Germany. ORCID
Martin Stanulla: Department of Pediatrics, University Medical Center Hannover, Hannover, Germany.
Justus Duyster: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
Peter Bronsert: Institute of Surgical Pathology, University of Freiburg Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Michael C Bassik: Department of Genetics, Stanford University School of Medicine, Stanford, CA.
Michael L Cleary: Department of Pathology, Stanford University School of Medicine, Stanford, CA.
Oliver Schilling: Institute for Molecular Medicine and Cell Research, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Merja Heinäniemi: Institute of Biomedicine, School of Medicine, University of Eastern Finland, Kuopio, Finland. ORCID
Jesús Duque-Afonso: Department of Hematology and Oncology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany. ORCID
ABSTRACT: The t(1;19) translocation, encoding the oncogenic fusion protein E2A (TCF3)-PBX1, is involved in acute lymphoblastic leukemia (ALL) and associated with a pre-B-cell receptor (preBCR+) phenotype. Relapse in patients with E2A-PBX1+ ALL frequently occurs in the central nervous system (CNS). Therefore, there is a medical need for the identification of CNS active regimens for the treatment of E2A-PBX1+/preBCR+ ALL. Using unbiased short hairpin RNA (shRNA) library screening approaches, we identified Bruton tyrosine kinase (BTK) as a key gene involved in both proliferation and dasatinib sensitivity of E2A-PBX1+/preBCR+ ALL. Depletion of BTK by shRNAs resulted in decreased proliferation of dasatinib-treated E2A-PBX1+/preBCR+ cells compared with control-transduced cells. Moreover, the combination of dasatinib with BTK inhibitors (BTKi; ibrutinib, acalabrutinib, or zanubrutinib) significantly decreased E2A-PBX1+/preBCR+ human and murine cell proliferation, reduced phospholipase C gamma 2 (PLCG2) and BTK phosphorylation and total protein levels and increased disease-free survival of mice in secondary transplantation assays, particularly reducing CNS-leukemic infiltration. Hence, dasatinib with ibrutinib reduced pPLCG2 and pBTK in primary ALL patient samples, including E2A-PBX1+ ALLs. In summary, genetic depletion and pharmacological inhibition of BTK increase dasatinib effects in human and mouse with E2A-PBX1+/preBCR+ ALL across most of performed assays, with the combination of dasatinib and BTKi proving effective in reducing CNS infiltration of E2A-PBX1+/preBCR+ ALL cells in vivo.
References
Blood. 2008 Aug 15;112(4):1005-12
[PMID: 18477770]
