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

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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.

LanguageEnglish
Pages215-225
JournalEBioMedicine
Volume39
DOIs
Publication statusPublished - Jan 2019

Cite this

@article{8613decbe0b34b4c84f22b3c4af80c71,
title = "Splicing modulation as novel therapeutic strategy against diffuse malignant peritoneal mesothelioma",
abstract = "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.",
author = "Rocco Sciarrillo and Anna Wojtuszkiewicz and {El Hassouni}, Btissame and Niccola Funel and Paolo Gandellini and Tonny Lagerweij and Silvia Buonamici and Maxime Blijlevens and {Zeeuw van der Laan}, {Eveline A} and Nadia Zaffaroni and Marcello Deraco and Shigeki Kusamura and Tom W{\"u}rdinger and Peters, {Godefridus J} and Molthoff, {Carla F M} and Gerrit Jansen and Kaspers, {Gertjan J L} and Jacqueline Cloos and Elisa Giovannetti",
note = "Copyright {\circledC} 2018. Published by Elsevier B.V.",
year = "2019",
month = "1",
doi = "10.1016/j.ebiom.2018.12.025",
language = "English",
volume = "39",
pages = "215--225",
journal = "EBioMedicine",
issn = "2352-3964",
publisher = "Elsevier BV",

}

Splicing modulation as novel therapeutic strategy against diffuse malignant peritoneal mesothelioma. / Sciarrillo, Rocco; Wojtuszkiewicz, Anna; El Hassouni, Btissame; Funel, Niccola; Gandellini, Paolo; Lagerweij, Tonny; Buonamici, Silvia; Blijlevens, Maxime; Zeeuw van der Laan, Eveline A; Zaffaroni, Nadia; Deraco, Marcello; Kusamura, Shigeki; Würdinger, Tom; Peters, Godefridus J; Molthoff, Carla F M; Jansen, Gerrit; Kaspers, Gertjan J L; Cloos, Jacqueline; Giovannetti, Elisa.

In: EBioMedicine, Vol. 39, 01.2019, p. 215-225.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Splicing modulation as novel therapeutic strategy against diffuse malignant peritoneal mesothelioma

AU - Sciarrillo, Rocco

AU - Wojtuszkiewicz, Anna

AU - El Hassouni, Btissame

AU - Funel, Niccola

AU - Gandellini, Paolo

AU - Lagerweij, Tonny

AU - Buonamici, Silvia

AU - Blijlevens, Maxime

AU - Zeeuw van der Laan, Eveline A

AU - Zaffaroni, Nadia

AU - Deraco, Marcello

AU - Kusamura, Shigeki

AU - Würdinger, Tom

AU - Peters, Godefridus J

AU - Molthoff, Carla F M

AU - Jansen, Gerrit

AU - Kaspers, Gertjan J L

AU - Cloos, Jacqueline

AU - Giovannetti, Elisa

N1 - Copyright © 2018. Published by Elsevier B.V.

PY - 2019/1

Y1 - 2019/1

N2 - 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.

AB - 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.

U2 - 10.1016/j.ebiom.2018.12.025

DO - 10.1016/j.ebiom.2018.12.025

M3 - Article

VL - 39

SP - 215

EP - 225

JO - EBioMedicine

T2 - EBioMedicine

JF - EBioMedicine

SN - 2352-3964

ER -