Synergistic drug interactions of the histone deacetylase inhibitor givinostat (ITF2357) in CRLF2-rearranged pediatric B-cell precursor acute lymphoblastic leukemia identified by high-throughput drug screening
Athanasios Oikonomou,
Titus Watrin,
Luigia Valsecchi,
Katerina Scharov,
Angela Maria Savino,
Julian Schliehe-Diecks,
Michela Bardini,
Grazia Fazio,
Silvia Bresolin,
Andrea Biondi,
Arndt Borkhardt,
Sanil Bhatia,
Giovanni Cazzaniga,
Chiara Palmi
Affiliations
Athanasios Oikonomou
Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy
Titus Watrin
Department of Paediatric Oncology, Haematology and Clinical Immunology, Heinrich-Heine University Dusseldorf, Medical Faculty, Düsseldorf, Germany
Luigia Valsecchi
Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy
Katerina Scharov
Department of Paediatric Oncology, Haematology and Clinical Immunology, Heinrich-Heine University Dusseldorf, Medical Faculty, Düsseldorf, Germany
Angela Maria Savino
Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy; School of Medicine and Surgery, University of Milano-Bicocca, Italy
Julian Schliehe-Diecks
Department of Paediatric Oncology, Haematology and Clinical Immunology, Heinrich-Heine University Dusseldorf, Medical Faculty, Düsseldorf, Germany
Michela Bardini
Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy
Grazia Fazio
Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy
Silvia Bresolin
Pediatric Hematology, Oncology and Stem Cell Transplant Division, Women and Child Health Department, Padua University and Hospital, Padua, Italy; Onco-Hematology, Stem Cell Transplant and Gene Therapy, Istituto di Ricerca Pediatrica Foundation - Città della Speranza, Padua, Italy
Andrea Biondi
School of Medicine and Surgery, University of Milano-Bicocca, Italy; Pediatrics, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy
Arndt Borkhardt
Department of Paediatric Oncology, Haematology and Clinical Immunology, Heinrich-Heine University Dusseldorf, Medical Faculty, Düsseldorf, Germany
Sanil Bhatia
Department of Paediatric Oncology, Haematology and Clinical Immunology, Heinrich-Heine University Dusseldorf, Medical Faculty, Düsseldorf, Germany
Giovanni Cazzaniga
Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy; School of Medicine and Surgery, University of Milano-Bicocca, Italy; Corresponding author. Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Via Pergolesi, 33, 20900, Monza, (MB), Italy.
Chiara Palmi
Tettamanti Center, Fondazione IRCCS San Gerardo dei Tintori, Monza, Italy
Combining multiple drugs broadens the window of therapeutic opportunities and is crucial for diseases that are currently lacking fully curative treatments. A powerful emerging tool for selecting effective drugs and combinations is the high-throughput drug screening (HTP). The histone deacetylase inhibitor (HDACi) givinostat (ITF2357) has been shown to act effectively against CRLF2-rearranged pediatric B-cell precursor acute lymphoblastic leukemia (BCP-ALL), a subtype characterized by poor outcome and enriched in children with Down Syndrome, very fragile patients with a high susceptibility to treatment-related toxicity. The aim of this study is to investigate possible synergies with givinostat for these difficult-to-treat patients by performing HTP screening with a library of 174 drugs, either approved or in preclinical studies. By applying this approach to the CRLF2-r MHH-CALL-4 cell line, we identified 19 compounds with higher sensitivity in combination with givinostat compared to the single treatments. Next, the synergy between givinostat and the promising candidates was further validated in CRLF2r cell lines with a broad matrix of concentrations. The combinations with trametinib (MEKi) or venetoclax (BCL2i) were found to be the most effective and with the greatest synergy across three metrics (ZIP, HAS, Bliss). Their efficacy was confirmed in primary blasts treated ex vivo at concentration ranges with a safe profile on healthy cells. Finally, we described givinostat-induced modifications in gene expression of MAPK and BCL-2 family members, supporting the observed synergistic interactions. Overall, our study represents a model of drug repurposing strategy using HTP screening for identifying synergistic, efficient, and safe drug combinations.
