RBL1 (p107) functions as tumor suppressor in glioblastoma and small-cell pancreatic neuroendocrine carcinoma in Xenopus tropicalis.
Animals
Animals, Genetically Modified
CRISPR-Cas Systems
/ genetics
Carcinoma, Neuroendocrine
/ genetics
Carcinoma, Small Cell
/ genetics
Disease Models, Animal
Gene Editing
Glioblastoma
/ genetics
Humans
Pancreatic Neoplasms
/ genetics
Retinoblastoma-Like Protein p107
/ genetics
Signal Transduction
/ genetics
Xenopus
Xenopus Proteins
/ genetics
Pancreatic Neoplasms
Journal
Oncogene
ISSN: 1476-5594
Titre abrégé: Oncogene
Pays: England
ID NLM: 8711562
Informations de publication
Date de publication:
03 2020
03 2020
Historique:
received:
15
03
2019
accepted:
20
01
2020
revised:
13
01
2020
pubmed:
1
2
2020
medline:
15
12
2020
entrez:
1
2
2020
Statut:
ppublish
Résumé
Alterations of the retinoblastoma and/or the p53 signaling network are associated with specific cancers such as high-grade astrocytoma/glioblastoma, small-cell lung cancer (SCLC), choroid plexus tumors, and small-cell pancreatic neuroendocrine carcinoma (SC-PaNEC). However, the intricate functional redundancy between RB1 and the related pocket proteins RBL1/p107 and RBL2/p130 in suppressing tumorigenesis remains poorly understood. Here we performed lineage-restricted parallel inactivation of rb1 and rbl1 by multiplex CRISPR/Cas9 genome editing in the true diploid Xenopus tropicalis to gain insight into this in vivo redundancy. We show that while rb1 inactivation is sufficient to induce choroid plexus papilloma, combined rb1 and rbl1 inactivation is required and sufficient to drive SC-PaNEC, retinoblastoma and astrocytoma. Further, using a novel Li-Fraumeni syndrome-mimicking tp53 mutant X. tropicalis line, we demonstrate increased malignancy of rb1/rbl1-mutant glioma towards glioblastoma upon concomitant inactivation of tp53. Interestingly, although clinical SC-PaNEC samples are characterized by abnormal p53 expression or localization, in the current experimental models, the tp53 status had little effect on the establishment and growth of SC-PaNEC, but may rather be essential for maintaining chromosomal stability. SCLC was only rarely observed in our experimental setup, indicating requirement of additional or alternative oncogenic insults. In conclusion, we used CRISPR/Cas9 to delineate the tumor suppressor properties of Rbl1, generating new insights in the functional redundancy within the retinoblastoma protein family in suppressing neuroendocrine pancreatic cancer and glioma/glioblastoma.
Identifiants
pubmed: 32001819
doi: 10.1038/s41388-020-1173-z
pii: 10.1038/s41388-020-1173-z
doi:
Substances chimiques
Retinoblastoma-Like Protein p107
0
Xenopus Proteins
0
rbl1 protein, Xenopus
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2692-2706Références
Tong Y, Merino D, Nimmervoll B, Gupta K, Wang Y-D, Finkelstein D, et al. Cross-species genomics identifies TAF12, NFYC, and RAD54L as choroid plexus carcinoma oncogenes. Cancer Cell. 2015;27:712–27.
doi: 10.1016/j.ccell.2015.04.005
pubmed: 4458854
pmcid: 4458854
McLendon R, Friedman A, Bigner D, Van Meir EG, Brat DJ, Mastrogianakis GM, et al. Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature. 2008;455:1061–8.
doi: 10.1038/nature07385
Chow LML, Endersby R, Zhu X, Rankin S, Qu C, Zhang J, et al. Cooperativity within and among Pten, p53, and Rb pathways induces high-grade astrocytoma in adult brain. Cancer Cell. 2011;19:305–16.
doi: 10.1016/j.ccr.2011.01.039
pubmed: 3060664
pmcid: 3060664
Konukiewitz B, Schlitter AM, Jesinghaus M, Pfister D, Steiger K, Segler A, et al. Somatostatin receptor expression related to TP53 and RB1 alterations in pancreatic and extrapancreatic neuroendocrine neoplasms with a Ki67-index above 20%. Mod Pathol. 2017;30:587–98.
doi: 10.1038/modpathol.2016.217
George J, Lim JS, Jang SJ, Cun Y, Ozretić L, Kong G, et al. Comprehensive genomic profiles of small cell lung cancer. Nature. 2015;524:47–53.
doi: 10.1038/nature14664
pubmed: 4861069
pmcid: 4861069
Hann CL, Rudin CM. Management of small-cell lung cancer: incremental changes but hope for the future. Oncology. 2008;22:1486–92.
pubmed: 19133604
Basturk O, Yang Z, Tang LH, Hruban RH, Adsay V, McCall CM, et al. The high-grade (WHO G3) pancreatic neuroendocrine tumor category is morphologically and biologically heterogenous and includes both well differentiated and poorly differentiated neoplasms. Am J Surg Pathol. 2015;39:683–90.
doi: 10.1097/PAS.0000000000000408
pubmed: 4398606
pmcid: 4398606
Tabori U, Shlien A, Baskin B, Levitt S, Ray P, Alon N, et al. TP53 alterations determine clinical subgroups and survival of patients with choroid plexus tumors. J Clin Oncol. 2010;28:1995–2001.
doi: 10.1200/JCO.2009.26.8169
Chow RD, Guzman CD, Wang G, Schmidt F, Youngblood MW, Ye L, et al. AAV-mediated direct in vivo CRISPR screen identifies functional suppressors in glioblastoma. Nat Neurosci. 2017;20:1329–41.
doi: 10.1038/nn.4620
pubmed: 5614841
pmcid: 5614841
Dannenberg J-H, Schuijff L, Dekker M, Van Der Valk M, Te Riele H. Tissue-specific tumor suppressor activity of retinoblastoma gene homologs p107 and p130. 2004. https://doi.org/10.1101/gad.322004 .
Costa C, Paramio JM, Santos M. Skin tumors Rb(eing) uncovered. Front Oncol. 2013;3:307.
doi: 10.3389/fonc.2013.00307
pubmed: 3865458
pmcid: 3865458
Robanus-Maandag E, Dekker M, van der Valk M, Carrozza ML, Jeanny JC, Dannenberg JH, et al. p107 is a suppressor of retinoblastoma development in pRb-deficient mice. Genes Dev. 1998;12:1599–609.
doi: 10.1101/gad.12.11.1599
pubmed: 316874
pmcid: 316874
Naert T, Colpaert R, Van Nieuwenhuysen T, Dimitrakopoulou D, Leoen J, Haustraete J, et al. CRISPR/Cas9 mediated knockout of rb1 and rbl1 leads to rapid and penetrant retinoblastoma development in Xenopus tropicalis. Sci Rep. 2016;6. https://doi.org/10.1038/srep35264 .
Xiao A, Wu H, Pandolfi PP, Louis DN, Van Dyke T. Astrocyte inactivation of the pRb pathway predisposes mice to malignant astrocytoma development that is accelerated by PTEN mutation. Cancer Cell. 2002;1:157–68.
doi: 10.1016/S1535-6108(02)00029-6
Lu X, Magrane G, Yin C, Louis DN, Gray J, Van Dyke T. Selective inactivation of p53 facilitates mouse epithelial tumor progression without chromosomal instability. Mol Cell Biol. 2001;21:6017–30.
doi: 10.1128/MCB.21.17.6017-6030.2001
pubmed: 87319
pmcid: 87319
Naert T, Van Nieuwenhuysen T, Vleminckx K. TALENs and CRISPR/Cas9 fuel genetically engineered clinically relevant Xenopus tropicalis tumor models. Genesis. 2017;55:e23005.
doi: 10.1002/dvg.23005
Bougeard G, Renaux-Petel M, Flaman J-M, Charbonnier C, Fermey P, Belotti M, et al. Revisiting Li-fraumeni syndrome from TP53 mutation carriers. J Clin Oncol. 2015;33:2345–52.
doi: 10.1200/JCO.2014.59.5728
Donehower LA, Harvey M, Slagle BL, McArthur MJ, Montgomery CA, Butel JS, et al. Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature. 1992;356:215–21.
doi: 10.1038/356215a0
Ignatius MS, Hayes MN, Moore FE, Tang Q, Garcia SP, Blackburn PR, et al. tp53 deficiency causes a wide tumor spectrum and increases embryonal rhabdomyosarcoma metastasis in zebrafish. Elife. 2018;7. https://doi.org/10.7554/eLife.37202 .
Berghmans S, Murphey RD, Wienholds E, Neuberg D, Kutok JL, Fletcher CDM, et al. tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors. Proc Natl Acad Sci. 2005;102:407–12.
doi: 10.1073/pnas.0406252102
Manning AL, Benes C, Dyson NJ. Whole chromosome instability resulting from the synergistic effects of pRB and p53 inactivation. Oncogene. 2014;33:2487–94.
doi: 10.1038/onc.2013.201
Eischen CM. Genome stability requires p53. Cold Spring Harb Perspect Med. 2016;6:a026096.
doi: 10.1101/cshperspect.a026096
pubmed: 4888814
pmcid: 4888814
Naert T, Vleminckx K. CRISPR/Cas9 disease models in zebrafish and Xenopus: the genetic renaissance of fish and frogs. Drug Discov Today Technol. 2018;28:41–52.
doi: 10.1016/j.ddtec.2018.07.001
Boel A, Steyaert W, De Rocker N, Menten B, Callewaert B, De Paepe A, et al. BATCH-GE: batch analysis of next-generation sequencing data for genome editing assessment. Sci Rep. 2016;6:30330.
doi: 10.1038/srep30330
pubmed: 4962088
pmcid: 4962088
Neely HR, Guo J, Flowers EM, Criscitiello MF, Flajnik MF. ‘Double-duty’ conventional dendritic cells in the amphibian Xenopus as the prototype for antigen presentation to B cells. Eur J Immunol. 2018;48:430–40.
doi: 10.1002/eji.201747260
pubmed: 5844829
pmcid: 5844829
Ward JM, Tadesse-Heath L, Perkins SN, Chattopadhyay SK, Hursting SD, Morse HC. Splenic marginal zone B-cell and thymic T-cell lymphomas in p53-deficient mice. Lab Investig. 1999;79:3–14.
pubmed: 9952106
Jacks T, Remington L, Williams BO, Schmitt EM, Halachmi S, Bronson RT, et al. Tumor spectrum analysis in p53-mutant mice. Curr Biol. 1994;4:1–7.
doi: 10.1016/S0960-9822(00)00002-6
Maresch R, Mueller S, Veltkamp C, Öllinger R, Friedrich M, Heid I, et al. Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice. Nat Commun. 2016;7. https://doi.org/10.1038/ncomms10770 .
van Zoest ID, Heijmen PS, Cruijsen PMJM, Jenks BG. Dynamics of background adaptation in Xenopus laevis: role of catecholamines and melanophore-stimulating hormone. Gen Comp Endocrinol. 1989;76:19–28.
doi: 10.1016/0016-6480(89)90028-2
Marino S, Vooijs M, van Der Gulden H, Jonkers J, Berns A. Induction of medulloblastomas in p53-null mutant mice by somatic inactivation of Rb in the external granular layer cells of the cerebellum. Genes Dev. 2000;14:994–1004.
pubmed: 316543
pmcid: 316543
McEvoy J, Flores-Otero J, Zhang J, Nemeth K, Brennan R, Bradley C, et al. Coexpression of normally incompatible developmental pathways in retinoblastoma genesis. Cancer Cell. 2011;20:260–75.
doi: 10.1016/j.ccr.2011.07.005
pubmed: 3551581
pmcid: 3551581
Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, et al. The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathol. 2016;131:803–20.
doi: 10.1007/s00401-016-1545-1
Steed TC, Treiber JM, Patel K, Ramakrishnan V, Merk A, Smith AR, et al. Differential localization of glioblastoma subtype: implications on glioblastoma pathogenesis. Oncotarget. 2016;7:24899–907.
doi: 10.18632/oncotarget.8551
pubmed: 5041878
pmcid: 5041878
Schaffer BE, Park K-S, Yiu G, Conklin JF, Lin C, Burkhart DL, et al. Loss of p130 accelerates tumor development in a mouse model for human small-cell lung carcinoma. Cancer Res. 2010;70:3877–83.
doi: 10.1158/0008-5472.CAN-09-4228
pubmed: 2873158
pmcid: 2873158
Yachida S, Vakiani E, White CM, Zhong Y, Saunders T, Morgan R, et al. Small cell and large cell neuroendocrine carcinomas of the pancreas are genetically similar and distinct from well-differentiated pancreatic neuroendocrine tumors. Am J Surg Pathol. 2012;36:173–84.
doi: 10.1097/PAS.0b013e3182417d36
pubmed: 3261427
pmcid: 3261427
Abou-El-Ardat K, Seifert M, Becker K, Eisenreich S, Lehmann M, Hackmann K, et al. Comprehensive molecular characterization of multifocal glioblastoma proves its monoclonal origin and reveals novel insights into clonal evolution and heterogeneity of glioblastomas. Neuro Oncol. 2017;19:546–57.
doi: 10.1093/neuonc/now231
pubmed: 5464316
pmcid: 5464316
Wiedemeyer WR, Dunn IF, Quayle SN, Zhang J, Chheda MG, Dunn GP, et al. Pattern of retinoblastoma pathway inactivation dictates response to CDK4/6 inhibition in GBM. Proc Natl Acad Sci USA. 2010;107:11501–6.
doi: 10.1073/pnas.1001613107
Schmid RS, Simon JM, Vitucci M, McNeill RS, Bash RE, Werneke AM, et al. Core pathway mutations induce de-differentiation of murine astrocytes into glioblastoma stem cells that are sensitive to radiation but resistant to temozolomide. Neuro Oncol. 2016;18:962–73.
doi: 10.1093/neuonc/nov321
pubmed: 4896545
pmcid: 4896545
Liu F, Gong J, Huang W, Wang Z, Wang M, Yang J, et al. MicroRNA-106b-5p boosts glioma tumorigensis by targeting multiple tumor suppressor genes. Oncogene. 2014;33:4813–22.
doi: 10.1038/onc.2013.428
Vitucci M, Irvin DM, McNeill RS, Schmid RS, Simon JM, Dhruv HD, et al. Genomic profiles of low-grade murine gliomas evolve during progression to glioblastoma. Neuro Oncol. 2017;19:1237–47.
doi: 10.1093/neuonc/nox050
pubmed: 5570221
pmcid: 5570221
Wojton J, Chu Z, Mathsyaraja H, Meisen WH, Denton N, Kwon C-H, et al. Systemic delivery of SapC-DOPS has antiangiogenic and antitumor effects against glioblastoma. Mol Ther. 2013;21:1517–25.
doi: 10.1038/mt.2013.114
pubmed: 3734660
pmcid: 3734660
Vanderluit JL, Wylie CA, McClellan KA, Ghanem N, Fortin A, Callaghan S, et al. The retinoblastoma family member p107 regulates the rate of progenitor commitment to a neuronal fate. J Cell Biol. 2007;178:129–39.
doi: 10.1083/jcb.200703176
pubmed: 2064429
pmcid: 2064429
Pan D, Chen Y, Du Y, Ren Z, Li X, Hu B. Methylation of promoter of RBL1 enhances the radioresistance of three dimensional cultured carcinoma cells. Oncotarget. 2017;8:4422–35.
pubmed: 27779109
Harb G, Vasavada RC, Cobrinik D, Stewart AF. The retinoblastoma protein and its homolog p130 regulate the G1/S transition in pancreatic beta-cells. Diabetes. 2009;58:1852–62.
doi: 10.2337/db08-0759
pubmed: 2712776
pmcid: 2712776
Cai EP, Luk CT, Wu X, Schroer SA, Shi SY, Sivasubramaniyam T, et al. Rb and p107 are required for alpha cell survival, beta cell cycle control and glucagon-like peptide-1 action. Diabetologia. 2014;57:2555–65.
doi: 10.1007/s00125-014-3381-y
Vasavada RC, Cozar-Castellano I, Sipula D, Stewart AF. Tissue-specific deletion of the retinoblastoma protein in the pancreatic beta-cell has limited effects on beta-cell replication, mass, and function. Diabetes. 2007;56:57–64.
doi: 10.2337/db06-0517
Glenn ST, Jones CA, Sexton S, LeVea CM, Caraker SM, Hajduczok G, et al. Conditional deletion of p53 and Rb in the renin-expressing compartment of the pancreas leads to a highly penetrant metastatic pancreatic neuroendocrine carcinoma. Oncogene. 2014;33:5706–15.
doi: 10.1038/onc.2013.514
Solin SL, Shive HR, Woolard KD, Essner JJ, McGrail M. Rapid tumor induction in zebrafish by TALEN-mediated somatic inactivation of the retinoblastoma1 tumor suppressor rb1. Sci Rep. 2015;5:13745.
doi: 10.1038/srep13745
pubmed: 4642565
pmcid: 4642565
Shim J, Choi J-H, Park M-H, Kim H, Kim JH, Kim S-Y, et al. Development of zebrafish medulloblastoma-like PNET model by TALEN-mediated somatic gene inactivation. Oncotarget. 2017;8:55280–97.
pubmed: 5589658
pmcid: 5589658
Jiang Z, Deng T, Jones R, Li H, Herschkowitz JI, Liu JC, et al. Rb deletion in mouse mammary progenitors induces luminal-B or basal-like/EMT tumor subtypes depending on p53 status. J Clin Investig. 2010;120:3296–309.
doi: 10.1172/JCI41490
Shi Z, Xin H, Tian D, Lian J, Wang J, Liu G, et al. Modeling human point mutation diseases in Xenopus tropicalis with a modified CRISPR/Cas9 system. FASEB J. 2019;33:6962–8.
doi: 10.1096/fj.201802661R
Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, et al. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature. 2019;576:149–57
Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin EV, et al. RNA-guided DNA insertion with CRISPR-associated transposases. Science. 2019;365:48–53.
doi: 10.1126/science.aax9181
pubmed: 6659118
pmcid: 6659118
Naert T, Nieuwenhuysen T Van, Demuynck S, Grande S de, Przybyl J, Vuylsteke M, et al. CRISPR-NSID: an in vivo CRISPR/Cas9 negative selection screen reveals EZH2 as a druggable dependency factor in a genetic desmoid tumor model. 2019. https://www.biorxiv.org/content/10.1101/595769v1 .
Moreno-Mateos MA, Vejnar CE, Beaudoin J-D, Fernandez JP, Mis EK, Khokha MK, et al. CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo. Nat Methods. 2015;12:982–8.
doi: 10.1038/nmeth.3543
pubmed: 4589495
pmcid: 4589495
Naert T, Vleminckx K. Methods for CRISPR/Cas9 Xenopus tropicalis Tissue-Specific Multiplex Genome Engineering. Methods Mol Biol. 2018;1865:33–54.
doi: 10.1007/978-1-4939-8784-9_3
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002;3:research0034.1.
doi: 10.1186/gb-2002-3-7-research0034