Abstract
Splicing alterations are common in diseases such as cancer, where mutations in splicing factor genes are frequently responsible for aberrant splicing. Here we present an alternative mechanism for splicing regulation in T-cell acute lymphoblastic leukemia (T-ALL) that involves posttranslational stabilization of the splicing machinery via deubiquitination. We demonstrate there are extensive exon skipping changes in disease, affecting proteasomal subunits, cell-cycle regulators, and the RNA machinery. We present that the serine/arginine-rich splicing factors (SRSF), controlling exon skipping, are critical for leukemia cell survival. The ubiquitin-specific pepti-dase 7 (USP7) regulates SRSF6 protein levels via active deubiquitination, and USP7 inhibition alters the exon skipping pattern and blocks T-ALL growth. The splicing inhibitor H3B-8800 affects splicing of proteasomal transcripts and proteasome activity and acts synergistically with proteasome inhibitors in inhibiting T-ALL growth. Our study provides the proof-of-principle for regulation of splicing factors via deubiquitination and suggests new therapeutic modalities in T-ALL. Significance: Our study provides a new proof-of-principle for posttranslational regulation of splicing factors independently of mutations in aggressive T-cell leukemia. It further suggests a new drug combination of splicing and proteasomal inhibitors, a concept that might apply to other diseases with or without mutations affecting the splicing machinery.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1388-1410 |
| Number of pages | 23 |
| Journal | Cancer discovery |
| Volume | 10 |
| Issue number | 9 |
| DOIs | |
| State | Published - Sep 2020 |
Funding
The Ntziachristos laboratory is or has been supported by the NCI (R00CA188293 and R01CA248770), the National Science Foundation, the Hartwell Foundation, a Gilead Research Scholarship, the American Society of Hematology, the Leukemia Research Foundation, the St. Baldrick’s Foundation, the H Foundation, the Gabrielle’s Angel Foundation, the Elsa Pardee Foundation, and the Zell Foundation. B.T. Guttierrez Diaz has been supported by the Fulbright Foundation and The Graduate School of Northwestern University. D.R. Amici was supported by 5T32GM008152-33. Proteomics services were performed by the Northwestern Proteomics Core Facility, supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center, instrumentation award (S10OD025194) from NIH Office of Director, and the National Resource for Translational and Developmental Proteomics supported by P41 GM108569. A. Tsirigos is supported by the American Cancer Society (RSG-15-189-01-RMC), St. Baldrick’s foundation (581357), and NCI/NIH P01CA229086-01A1. P. Van Vlierberghe is supported by funding from the European Research Council (ERC-StG-639784). We thank the NYU Langone Genome Technology Center (GTC) for PacBio Sequel sequencing, acquired with the Shared Instrumentation Grant 1S10OD023423-01 from NIH. We also thank the Applied Bioinformatics Laboratories (ABL) for providing bioinformatics support and helping with the analysis and The Ntziachristos laboratory is or has been supported by the NCI (R00CA188293 and R01CA248770), the National Science Foundation, the Hartwell Foundation, a Gilead Research Scholarship, the American Society of Hematology, the Leukemia Research Foundation, the St. Baldrick?s Foundation, the H Foundation, the Gabrielle?s Angel Foun-dation, the Elsa Pardee Foundation, and the Zell Foundation. B.T. Guttierrez Diaz has been supported by the Fulbright Foundation and The Graduate School of Northwestern University. D.R. Amici was supported by 5T32GM008152-33. Proteomics services were performed by the Northwestern Proteomics Core Facility, supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center, instrumentation award (S10OD025194) from NIH Office of Director, and the National Resource for Translational and Developmental Proteomics supported by P41 GM108569. A. Tsirigos is supported by the American Cancer Society (RSG-15-189-01-RMC), St. Baldrick?s foundation (581357), and NCI/NIH P01CA229086-01A1. P. Van Vlierberghe is supported by funding from the European Research Council (ERC-StG-639784). We thank the NYU Langone Genome Technology Center (GTC) for PacBio Sequel sequencing, acquired with the Shared Instrumentation Grant 1S10OD023423-01 from NIH. We also thank the Applied Bioinformatics Laboratories (ABL) for providing bioinformatics support and helping with the analysis and interpretation of the data. GTC and ABL are shared resources partially supported by the NYU Cancer Center Support Grant P30CA016087 at the Laura and Isaac Perlmutter Cancer Center. This work has used computing resources at the NYU School of Medicine High Perfor-mance Computing Facility. We want to thank Dr. Issam Ben Sahra and laboratory and the Department of Biochemistry and Molecular Genetics for helping with reagents and equipment used in the study as well as all the members of the Ntziachristos laboratory for their comments and critical review of the manuscript. interpretation of the data. GTC and ABL are shared resources partially supported by the NYU Cancer Center Support Grant P30CA016087 at the Laura and Isaac Perlmutter Cancer Center. This work has used computing resources at the NYU School of Medicine High Performance Computing Facility. We want to thank Dr. Issam Ben Sahra and laboratory and the Department of Biochemistry and Molecular Genetics for helping with reagents and equipment used in the study as well as all the members of the Ntziachristos laboratory for their comments and critical review of the manuscript.
ASJC Scopus subject areas
- Oncology
