Elys deficiency constrains Kras-driven tumour burden by amplifying oncogenic stress

The nucleoporin ELYS, encoded by AHCTF1, is a large multifunctional protein with essential roles in nuclear pore assembly and mitosis. Using a zebrafish model of hepatocellular carcinoma, in which the expression of an inducible mutant kras transgene (krasG12V) drives hepatocyte-specific hyperplasia and liver enlargement, we show that reducing ahctf1 gene dosage by 50% markedly shrinks tumour burden, while non-hyperplastic tissues are unaffected. We demonstrate that ahctf1 heterozygosity impairs nuclear pore formation, mitotic spindle assembly and chromosome segregation, leading to DNA damage and activation of TP53-dependent and independent mechanisms of cell death and cell cycle arrest. This selective vulnerability of cancer cells to mild disruption of Elys function uncovers a novel synthetic lethal interaction between ahctf1 and oncogenic kras that could be exploited therapeutically. Heterozygous expression of both ahctf1 and ranbp2, or treatment of heterozygous ahctf1 larvae with the nucleocytoplasmic transport inhibitor, Selinexor, completely blocked krasG12V-driven hepatocyte hyperplasia, revealing promising avenues for combinatorial treatments.

[1]  A. Pandiella,et al.  Proteolysis targeting chimeras (PROTACs) in cancer therapy , 2020, Journal of Experimental & Clinical Cancer Research.

[2]  Jenifer B. Kaplan,et al.  PEARL-seq: A Photoaffinity Platform for the Analysis of Small Molecule-RNA Interactions. , 2020, ACS chemical biology.

[3]  P. Currie,et al.  Cellular rescue in a zebrafish model of congenital muscular dystrophy type 1A , 2019, npj Regenerative Medicine.

[4]  Shawn C. Little,et al.  Chromatin targeting of nuclear pore proteins induces chromatin decondensation , 2019, The Journal of cell biology.

[5]  Aviad Tsherniak,et al.  Extracting Biological Insights from the Project Achilles Genome-Scale CRISPR Screens in Cancer Cell Lines , 2019, bioRxiv.

[6]  D. Jans,et al.  Inhibitors of nuclear transport. , 2019, Current opinion in cell biology.

[7]  H. Kurumizaka,et al.  Structural and biochemical analyses of the nuclear pore complex component ELYS identify residues responsible for nucleosome binding , 2019, Communications Biology.

[8]  Z. Gong,et al.  Transcriptomic analyses of oncogenic hepatocytes reveal common and different molecular pathways of hepatocarcinogenesis in different developmental stages and genders in krasG12V transgenic zebrafish. , 2019, Biochemical and biophysical research communications.

[9]  S. Lowe,et al.  Nuclear Pores Promote Lethal Prostate Cancer by Increasing POM121-Driven E2F1, MYC, and AR Nuclear Import , 2018, Cell.

[10]  L. Lai,et al.  Synthetic lethality in drug development: the dawn is coming. , 2018, Future medicinal chemistry.

[11]  P. Schirmacher,et al.  Alterations of the nuclear transport system in hepatocellular carcinoma - New basis for therapeutic strategies. , 2017, Journal of hepatology.

[12]  Ann E. Sizemore,et al.  Computational correction of copy-number effect improves specificity of CRISPR-Cas9 essentiality screens in cancer cells , 2017, Nature Genetics.

[13]  Alan Ashworth,et al.  PARP inhibitors: Synthetic lethality in the clinic , 2017, Science.

[14]  Eric S. Lander,et al.  Gene Essentiality Profiling Reveals Gene Networks and Synthetic Lethal Interactions with Oncogenic Ras , 2017, Cell.

[15]  M. Beck,et al.  The nuclear pore complex: understanding its function through structural insight , 2016, Nature Reviews Molecular Cell Biology.

[16]  R. Bernards,et al.  A Vulnerability of a Subset of Colon Cancers with Potential Clinical Utility , 2016, Cell.

[17]  J. Zucman‐Rossi,et al.  Genetic Landscape and Biomarkers of Hepatocellular Carcinoma. , 2015, Gastroenterology.

[18]  S. Gery,et al.  KPT-330 inhibitor of XPO1-mediated nuclear export has anti-proliferative activity in hepatocellular carcinoma , 2014, Cancer Chemotherapy and Pharmacology.

[19]  B. C. Low,et al.  Crosstalk of Ras and Rho: activation of RhoA abates Kras-induced liver tumorigenesis in transgenic zebrafish models , 2014, Oncogene.

[20]  Weiling Zheng,et al.  Xmrk, Kras and Myc Transgenic Zebrafish Liver Cancer Models Share Molecular Signatures with Subsets of Human Hepatocellular Carcinoma , 2014, PloS one.

[21]  S. Grimmond,et al.  Minor class splicing shapes the zebrafish transcriptome during development , 2014, Proceedings of the National Academy of Sciences.

[22]  Juan Carlos González-Sánchez,et al.  The nucleoporin MEL-28 promotes RanGTP-dependent γ-tubulin recruitment and microtubule nucleation in mitotic spindle formation , 2014, Nature Communications.

[23]  R. Hill,et al.  Targeting nucleocytoplasmic transport in cancer therapy , 2013, Oncotarget.

[24]  C. Hashizume,et al.  Down-modulation of nucleoporin RanBP2/Nup358 impaired chromosomal alignment and induced mitotic catastrophe , 2013, Cell Death and Disease.

[25]  John R. Yates,et al.  Identification of Long-Lived Proteins Reveals Exceptional Stability of Essential Cellular Structures , 2013, Cell.

[26]  Benjamin E. Gross,et al.  Integrative Analysis of Complex Cancer Genomics and Clinical Profiles Using the cBioPortal , 2013, Science Signaling.

[27]  T. Schwartz,et al.  Structural and functional studies of the 252 kDa nucleoporin ELYS reveal distinct roles for its three tethered domains. , 2013, Structure.

[28]  Seok-Hyung Kim,et al.  Autophagy Induction Is a Tor- and Tp53-Independent Cell Survival Response in a Zebrafish Model of Disrupted Ribosome Biogenesis , 2013, PLoS genetics.

[29]  A. Strasser,et al.  DNA damage-induced primordial follicle oocyte apoptosis and loss of fertility require TAp63-mediated induction of Puma and Noxa. , 2012, Molecular cell.

[30]  T. Schwartz,et al.  Molecular basis for Nup37 and ELY5/ELYS recruitment to the nuclear pore complex , 2012, Proceedings of the National Academy of Sciences.

[31]  B. Paw,et al.  Mutation mapping and identification by whole-genome sequencing , 2012, Genome research.

[32]  Benjamin E. Gross,et al.  The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. , 2012, Cancer discovery.

[33]  W. Driever,et al.  Fast Homozygosity Mapping and Identification of a Zebrafish ENU-Induced Mutation by Whole-Genome Sequencing , 2012, PloS one.

[34]  B. Fahrenkrog,et al.  Nucleoporins: leaving the nuclear pore complex for a successful mitosis. , 2011, Cellular signalling.

[35]  G Melino,et al.  p63 and p73, the ancestors of p53. , 2010, Cold Spring Harbor perspectives in biology.

[36]  B. Fontoura,et al.  The Nup107-160 complex and γ-TuRC regulate microtubule polymerization at kinetochores , 2010, Nature Cell Biology.

[37]  Michael J. Emanuele,et al.  A Genome-wide RNAi Screen Identifies Multiple Synthetic Lethal Interactions with the Ras Oncogene , 2009, Cell.

[38]  L. Zon,et al.  Abnormal nuclear pore formation triggers apoptosis in the intestinal epithelium of elys-deficient zebrafish. , 2009, Gastroenterology.

[39]  U. Kutay,et al.  Orchestrating nuclear envelope disassembly and reassembly during mitosis , 2009, Nature Reviews Molecular Cell Biology.

[40]  A. Moorman,et al.  Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data , 2009, Nucleic acids research.

[41]  M. D'Angelo,et al.  Age-Dependent Deterioration of Nuclear Pore Complexes Causes a Loss of Nuclear Integrity in Postmitotic Cells , 2009, Cell.

[42]  M. Pack,et al.  Mutation of the Zebrafish Nucleoporin elys Sensitizes Tissue Progenitors to Replication Stress , 2008, PLoS genetics.

[43]  Thorsten Wohland,et al.  Requirement of vasculogenesis and blood circulation in late stages of liver growth in zebrafish , 2008, BMC Developmental Biology.

[44]  A. Harel,et al.  Capture of AT-rich chromatin by ELYS recruits POM121 and NDC1 to initiate nuclear pore assembly. , 2008, Molecular biology of the cell.

[45]  J. Swedlow,et al.  ELYS/MEL-28 Chromatin Association Coordinates Nuclear Pore Complex Assembly and Replication Licensing , 2007, Current Biology.

[46]  S. Elledge,et al.  Non-Oncogene Addiction and the Stress Phenotype of Cancer Cells , 2007, Cell.

[47]  M. Hetzer,et al.  MEL‐28/ELYS is required for the recruitment of nucleoporins to chromatin and postmitotic nuclear pore complex assembly , 2007, EMBO reports.

[48]  S. Briggs,et al.  ELYS is a dual nucleoporin/kinetochore protein required for nuclear pore assembly and proper cell division , 2006, Proceedings of the National Academy of Sciences.

[49]  A. Yang,et al.  p63 protects the female germ line during meiotic arrest , 2006, Nature.

[50]  Samantha G. Zeitlin,et al.  The Nup107-160 nucleoporin complex is required for correct bipolar spindle assembly. , 2006, Molecular biology of the cell.

[51]  D. Stainier,et al.  Mesodermal Wnt2b signalling positively regulates liver specification , 2006, Nature.

[52]  S. Thorgeirsson,et al.  Ubiquitous activation of Ras and Jak/Stat pathways in human HCC. , 2006, Gastroenterology.

[53]  L. Zon,et al.  tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[54]  H. Kiyonari,et al.  Targeted disruption of the mouse ELYS gene results in embryonic death at peri‐implantation development , 2004, Genes to cells : devoted to molecular & cellular mechanisms.

[55]  M. Dasso,et al.  The RanGAP1-RanBP2 Complex Is Essential for Microtubule-Kinetochore Interactions In Vivo , 2004, Current Biology.

[56]  Pamela A. Silver,et al.  Nuclear transport and cancer: from mechanism to intervention , 2004, Nature Reviews Cancer.

[57]  J. B. Rattner,et al.  Nup358 integrates nuclear envelope breakdown with kinetochore assembly , 2003, The Journal of cell biology.

[58]  G Melino,et al.  The p53/p63/p73 family of transcription factors: overlapping and distinct functions. , 2000, Journal of cell science.

[59]  C. Nüsslein-Volhard,et al.  Mutations affecting the cardiovascular system and other internal organs in zebrafish. , 1996, Development.

[60]  C. Nüsslein-Volhard,et al.  Mutations affecting pigmentation and shape of the adult zebrafish , 1996, Development Genes and Evolution.