The Splicing Factor DHX38/PRP16 is Required for Ovarian Clear Cell Carcinoma Tumorigenesis, As Revealed by a CRISPR-Cas9 Screen

Overview

Journal FEBS Open Bio

Specialty Biology

Date 2021 Dec 29

PMID 34965029

Citations 2

Authors

Brandon Cona

Tomoatsu Hayashi

Ai Yamada

Naomi Shimizu

Naoko Yokota

Ryuichiro Nakato

Katsuhiko Shirahige

Tetsu Akiyama

Affiliations

Soon will be listed here.

Abstract

Certain cancers, such as ovarian clear cell carcinoma (OCCC), display high levels of genetic variation between patients, making it difficult to develop effective therapies. In order to identify novel genes critical to OCCC growth, we carried out a comprehensive CRISPR-Cas9 knockout screen against cell growth using an OCCC cell line and a normal ovarian surface epithelium cell line. We identified the gene encoding DHX38/PRP16, an ATP-dependent RNA helicase involved in splicing, as critical for the growth and tumorigenesis of OCCC. DHX38/PRP16 knockdown in OCCC cells, but not normal cells, induces apoptosis and impairs OCCC tumorigenesis in a mouse model. Our results suggest that DHX38/PRP16 may play a role in OCCC tumorigenesis and could potentially be a promising therapeutic target.

Citing Articles

Knockout of Causes Inner Ear Developmental Defects in Zebrafish.

Ren M, Chen X, Dai L, Tu J, Hu H, Sun X Biomedicines. 2025; 13(1).

PMID: 39857604 PMC: 11760894. DOI: 10.3390/biomedicines13010020.

New insights into pathogen-mediated modulation of host RNA splicing.

Gao C, Dong S Stress Biol. 2023; 2(1):34.

PMID: 37676360 PMC: 10442024. DOI: 10.1007/s44154-022-00053-2.

Multimodal CRISPR perturbations of GWAS loci associated with coronary artery disease in vascular endothelial cells.

Wunnemann F, Fotsing Tadjo T, Beaudoin M, Lalonde S, Lo K, Kleinstiver B PLoS Genet. 2023; 19(3):e1010680.

PMID: 36928188 PMC: 10047545. DOI: 10.1371/journal.pgen.1010680.

References

Iida Y, Okamoto A, Hollis R, Gourley C, Herrington C . Clear cell carcinoma of the ovary: a clinical and molecular perspective. Int J Gynecol Cancer. 2020; 31(4):605-616. DOI: 10.1136/ijgc-2020-001656. View

Chen L, Chen J, Huang Y, Gu Y, Qiu J, Qian H . The Augmented R-Loop Is a Unifying Mechanism for Myelodysplastic Syndromes Induced by High-Risk Splicing Factor Mutations. Mol Cell. 2018; 69(3):412-425.e6. PMC: 5957072. DOI: 10.1016/j.molcel.2017.12.029. View

Michels B, Mosa M, Streibl B, Zhan T, Menche C, Abou-El-Ardat K . Pooled In Vitro and In Vivo CRISPR-Cas9 Screening Identifies Tumor Suppressors in Human Colon Organoids. Cell Stem Cell. 2020; 26(5):782-792.e7. DOI: 10.1016/j.stem.2020.04.003. View

Wang Y, Guthrie C . PRP16, a DEAH-box RNA helicase, is recruited to the spliceosome primarily via its nonconserved N-terminal domain. RNA. 1998; 4(10):1216-29. PMC: 1369694. View

Mi H, Dong Q, Muruganujan A, Gaudet P, Lewis S, Thomas P . PANTHER version 7: improved phylogenetic trees, orthologs and collaboration with the Gene Ontology Consortium. Nucleic Acids Res. 2009; 38(Database issue):D204-10. PMC: 2808919. DOI: 10.1093/nar/gkp1019. View

Sugiyama T, Kamura T, Kigawa J, Terakawa N, Kikuchi Y, Kita T . Clinical characteristics of clear cell carcinoma of the ovary: a distinct histologic type with poor prognosis and resistance to platinum-based chemotherapy. Cancer. 2000; 88(11):2584-9. View

Wilkinson M, Fica S, Galej W, Norman C, Newman A, Nagai K . Postcatalytic spliceosome structure reveals mechanism of 3'-splice site selection. Science. 2017; 358(6368):1283-1288. PMC: 5808836. DOI: 10.1126/science.aar3729. View

Hart T, Chandrashekhar M, Aregger M, Steinhart Z, Brown K, MacLeod G . High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities. Cell. 2015; 163(6):1515-26. DOI: 10.1016/j.cell.2015.11.015. View

Saijo S, Kuwano Y, Masuda K, Nishikawa T, Rokutan K, Nishida K . Serine/arginine-rich splicing factor 7 regulates p21-dependent growth arrest in colon cancer cells. J Med Invest. 2016; 63(3-4):219-26. DOI: 10.2152/jmi.63.219. View

10.

Zhan T, Rindtorff N, Betge J, Ebert M, Boutros M . CRISPR/Cas9 for cancer research and therapy. Semin Cancer Biol. 2018; 55:106-119. DOI: 10.1016/j.semcancer.2018.04.001. View

11.

Thomas P, Campbell M, Kejariwal A, Mi H, Karlak B, Daverman R . PANTHER: a library of protein families and subfamilies indexed by function. Genome Res. 2003; 13(9):2129-41. PMC: 403709. DOI: 10.1101/gr.772403. View

12.

Zhou Z, REED R . Human homologs of yeast prp16 and prp17 reveal conservation of the mechanism for catalytic step II of pre-mRNA splicing. EMBO J. 1998; 17(7):2095-106. PMC: 1170554. DOI: 10.1093/emboj/17.7.2095. View

13.

Kedzierska H, Piekielko-Witkowska A . Splicing factors of SR and hnRNP families as regulators of apoptosis in cancer. Cancer Lett. 2017; 396:53-65. DOI: 10.1016/j.canlet.2017.03.013. View

14.

Gadducci A, Multinu F, Cosio S, Carinelli S, Ghioni M, Aletti G . Clear cell carcinoma of the ovary: Epidemiology, pathological and biological features, treatment options and clinical outcomes. Gynecol Oncol. 2021; 162(3):741-750. DOI: 10.1016/j.ygyno.2021.06.033. View

15.

Yang J, Nie J, Ma X, Wei Y, Peng Y, Wei X . Targeting PI3K in cancer: mechanisms and advances in clinical trials. Mol Cancer. 2019; 18(1):26. PMC: 6379961. DOI: 10.1186/s12943-019-0954-x. View

16.

Fuller-Pace F . DEAD box RNA helicase functions in cancer. RNA Biol. 2013; 10(1):121-32. PMC: 3590229. DOI: 10.4161/rna.23312. View

17.

Schwer B, Guthrie C . PRP16 is an RNA-dependent ATPase that interacts transiently with the spliceosome. Nature. 1991; 349(6309):494-9. DOI: 10.1038/349494a0. View

18.

Fraile J, Manchado E, Lujambio A, Quesada V, Campos-Iglesias D, Webb T . USP39 Deubiquitinase Is Essential for Oncogene-driven Cancer. J Biol Chem. 2017; 292(10):4164-4175. PMC: 5354494. DOI: 10.1074/jbc.M116.762757. View

19.

Kurokawa K, Akaike Y, Masuda K, Kuwano Y, Nishida K, Yamagishi N . Downregulation of serine/arginine-rich splicing factor 3 induces G1 cell cycle arrest and apoptosis in colon cancer cells. Oncogene. 2013; 33(11):1407-17. DOI: 10.1038/onc.2013.86. View

20.

Bonnal S, Lopez-Oreja I, Valcarcel J . Roles and mechanisms of alternative splicing in cancer - implications for care. Nat Rev Clin Oncol. 2020; 17(8):457-474. DOI: 10.1038/s41571-020-0350-x. View