Splicing modulation as novel therapeutic strategy against diffuse malignant peritoneal mesothelioma
Rocco Sciarrillo,
Anna Wojtuszkiewicz,
Btissame El Hassouni,
Niccola Funel,
Paolo Gandellini,
Tonny Lagerweij,
Silvia Buonamici,
Maxime Blijlevens,
Eveline A. Zeeuw van der Laan,
Nadia Zaffaroni,
Marcello Deraco,
Shigeki Kusamura,
Tom Würdinger,
Godefridus J. Peters,
Carla F.M. Molthoff,
Gerrit Jansen,
Gertjan J.L. Kaspers,
Jacqueline Cloos,
Elisa Giovannetti
Affiliations
Rocco Sciarrillo
Department of Pediatric Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands; Department of Hematology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands; Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Anna Wojtuszkiewicz
Department of Pediatric Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands; Department of Hematology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Btissame El Hassouni
Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Niccola Funel
Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, Italy; CNR-Nano, Institute of Nanoscience and Nanotechnology, Pisa, Italy
Paolo Gandellini
Department of Applied Research and Technological Development, Molecular Pharmacology Unit, Fondazione IRCCS - National Cancer Institute, Milano, Italy
Tonny Lagerweij
Department of Neurosurgery, Neuro-Oncology Research Group, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Silvia Buonamici
H3 Biomedicine, Inc., 300 Technology Square, FL 5, Cambridge, MA 02139, USA
Maxime Blijlevens
Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Eveline A. Zeeuw van der Laan
Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Nadia Zaffaroni
Department of Applied Research and Technological Development, Molecular Pharmacology Unit, Fondazione IRCCS - National Cancer Institute, Milano, Italy
Marcello Deraco
Peritoneal Malignancy Program, National Cancer Institute, Milano, Italy
Shigeki Kusamura
Peritoneal Malignancy Program, National Cancer Institute, Milano, Italy
Tom Würdinger
Department of Neurosurgery, Neuro-Oncology Research Group, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Godefridus J. Peters
Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Carla F.M. Molthoff
Department of Radiology & Nuclear Medicine, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Gerrit Jansen
Amsterdam Rheumatology and Immunology Center, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Gertjan J.L. Kaspers
Princes Maxima Center for Pediatric Oncology, Utrecht, the Netherlands
Jacqueline Cloos
Department of Pediatric Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands; Department of Hematology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands
Elisa Giovannetti
Department of Medical Oncology, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Amsterdam, the Netherlands; Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, Italy; CNR-Nano, Institute of Nanoscience and Nanotechnology, Pisa, Italy; Corresponding author at: Department of Medical Oncology, Cancer Center Amsterdam, Cancer Center Amsterdam, Amsterdam University Medical Center, location VUMC, Room 1.52, De Boelelaan 1117, Amsterdam 1081 HV, the Netherlands.
Introduction: Therapeutic options for diffuse malignant peritoneal mesothelioma (DMPM) are limited to surgery and locoregional chemotherapy. Despite improvements in survival rates, patients eventually succumb to disease progression. We investigated splicing deregulation both as molecular prognostic factor and potential novel target in DMPM, while we tested modulators of SF3b complex for antitumor activity. Methods: Tissue-microarrays of 64 DMPM specimens were subjected to immunohistochemical assessment of SF3B1 expression and correlation to clinical outcome. Two primary cell cultures were used for gene expression profiling and in vitro screening of SF3b modulators. Drug-induced splicing alterations affecting downstream cellular pathways were detected through RNA sequencing. Ultimately, we established bioluminescent orthotopic mouse models to test the efficacy of splicing modulation in vivo. Results: Spliceosomal genes are differentially upregulated in DMPM cells compared to normal tissues and high expression of SF3B1 correlated with poor clinical outcome in univariate and multivariate analysis. SF3b modulators (Pladienolide-B, E7107, Meayamycin-B) showed potent cytotoxic activity in vitro with IC50 values in the low nanomolar range. Differential splicing analysis of Pladienolide-B-treated cells revealed abundant alterations of transcripts involved in cell cycle, apoptosis and other oncogenic pathways. This was validated by RT-PCR and functional assays. E7107 demonstrated remarkable in vivo antitumor efficacy, with significant improvement of survival rates compared to vehicle-treated controls. Conclusions: SF3B1 emerged as a novel potential prognostic factor in DMPM. Splicing modulators markedly impair cancer cell viability, resulting also in potent antitumor activity in vivo. Our data designate splicing as a promising therapeutic target in DMPM. Keywords: Mesothelioma, Therapy, SF3b modulation, Splicing, RNA-sequencing
