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Molecular Cancer
Published in: Molecular Cancer 1/2015

Open Access 01-12-2015 | Research

Oncogenic KRAS promotes malignant brain tumors in zebrafish

Authors: Bensheng Ju, Wenbiao Chen, Brent A Orr, Jan M Spitsbergen, Sujuan Jia, Christopher J Eden, Hannah E Henson, Michael R Taylor

Published in: Molecular Cancer | Issue 1/2015

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Abstract

Background

Zebrafish have been used as a vertebrate model to study human cancers such as melanoma, rhabdomyosarcoma, liver cancer, and leukemia as well as for high-throughput screening of small molecules of therapeutic value. However, they are just emerging as a model for human brain tumors, which are among the most devastating and difficult to treat. In this study, we evaluated zebrafish as a brain tumor model by overexpressing a human version of oncogenic KRAS (KRASG12V).

Methods

Using zebrafish cytokeratin 5 (krt5) and glial fibrillary acidic protein (gfap) gene promoters, we activated Ras signaling in the zebrafish central nervous system (CNS) through transient and stable transgenic overexpression. Immunohistochemical analyses were performed to identify activated pathways in the resulting brain tumors. The effects of the MEK inhibitor U0126 on oncogenic KRAS were evaluated.

Results

We demonstrated that transient transgenic expression of KRASG12V in putative neural stem and/or progenitor cells induced brain tumorigenesis. When expressed under the control of the krt5 gene promoter, KRASG12V induced brain tumors in ventricular zones (VZ) at low frequency. The majority of other tumors were composed mostly of spindle and epithelioid cells, reminiscent of malignant peripheral nerve sheath tumors (MPNSTs). In contrast, when expressed under the control of the gfap gene promoter, KRASG12V induced brain tumors in both VZs and brain parenchyma at higher frequency. Immunohistochemical analyses indicated prominent activation of the canonical RAS-RAF-ERK pathway, variable activation of the mTOR pathway, but no activation of the PI3K-AKT pathway. In a krt5-derived stable and inducible transgenic line, expression of oncogenic KRAS resulted in skin hyperplasia, and the MEK inhibitor U0126 effectively suppressed this pro-proliferative effects. In a gfap-derived stable and inducible line, expression of oncogenic KRAS led to significantly increased mitotic index in the spinal cord.

Conclusions

Our studies demonstrate that zebrafish could be explored to study cellular origins and molecular mechanisms of brain tumorigenesis and could also be used as a platform for studying human oncogene function and for discovering oncogenic RAS inhibitors.
Appendix
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Literature
1.
Maher EA, Furnari FB, Bachoo RM, Rowitch DH, Louis DN, Cavenee WK, et al. Malignant glioma: genetics and biology of a grave matter. Genes Dev. 2001;15:1311–33.PubMedCrossRef
2.
Monje M, Mitra SS, Freret ME, Raveh TB, Kim J, Masek M, et al. Hedgehog-responsive candidate cell of origin for diffuse intrinsic pontine glioma. Proc Natl Acad Sci U S A. 2011;108:4453–8.PubMedCentralPubMedCrossRef
3.
Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature. 2008;455:1061–68.
4.
Holmen SL, Williams BO. Essential role for Ras signaling in glioblastoma maintenance. Cancer Res. 2005;65:8250–5.PubMedCrossRef
5.
Abel TW, Clark C, Bierie B, Chytil A, Aakre M, Gorska A, et al. GFAP-Cre-mediated activation of oncogenic K-ras results in expansion of the subventricular zone and infiltrating glioma. Mol Cancer Res. 2009;7:645–53.PubMedCentralPubMedCrossRef
6.
Holland EC, Celestino J, Dai C, Schaefer L, Sawaya RE, Fuller GN. Combined activation of Ras and Akt in neural progenitors induces glioblastoma formation in mice. Nat Genet. 2000;25:55–7.PubMedCrossRef
7.
Munoz DM, Tung T, Agnihotri S, Singh S, Guha A, Zadeh G, et al. Loss of p53 cooperates with K-ras activation to induce glioma formation in a region-independent manner. Glia. 2013;61:1862–72.PubMed
8.
White R, Rose K, Zon L. Zebrafish cancer: the state of the art and the path forward. Nat Rev Cancer. 2013;13:624–36.PubMedCrossRef
9.
Patton EE, Widlund HR, Kutok JL, Kopani KR, Amatruda JF, Murphey RD, et al. BRAF mutations are sufficient to promote nevi formation and cooperate with p53 in the genesis of melanoma. Curr Biol. 2005;15:249–54.PubMedCrossRef
10.
Park SW, Davison JM, Rhee J, Hruban RH, Maitra A, Leach SD. Oncogenic KRAS induces progenitor cell expansion and malignant transformation in zebrafish exocrine pancreas. Gastroenterology. 2008;134:2080–90.PubMedCentralPubMedCrossRef
11.
Langenau DM, Keefe MD, Storer NY, Guyon JR, Kutok JL, Le X, et al. Effects of RAS on the genesis of embryonal rhabdomyosarcoma. Genes Dev. 2007;21:1382–95.PubMedCentralPubMedCrossRef
12.
Nguyen AT, Emelyanov A, Koh CH, Spitsbergen JM, Lam SH, Mathavan S, et al. A high level of liver-specific expression of oncogenic Kras(V12) drives robust liver tumorigenesis in transgenic zebrafish. Dis Model Mech. 2011;4:801–13.PubMedCentralPubMedCrossRef
13.
Ju B. Activation of Sonic hedgehog signaling in neural progenitor cells promotes glioma development in the zebrafish optic pathway. Oncogenesis. 2014;3:e96.PubMedCentralPubMedCrossRef
14.
15.
Krushna Padhi B, Akimenko MA, Ekker M. Independent expansion of the keratin gene family in teleostean fish and mammals: an insight from phylogenetic analysis and radiation hybrid mapping of keratin genes in zebrafish. Gene. 2006;368:37–45.PubMedCrossRef
16.
Chua KL, Lim TM. Type I and type II cytokeratin cDNAs from the zebrafish (Danio rerio) and expression patterns during early development. Differentiation. 2000;66:31–41.PubMedCrossRef
17.
Lam CS, Marz M, Strahle U. gfap and nestin reporter lines reveal characteristics of neural progenitors in the adult zebrafish brain. Dev Dyn. 2009;238:475–86.PubMedCrossRef
18.
Winkler C, Schafer M, Duschl J, Schartl M, Volff JN. Functional divergence of two zebrafish midkine growth factors following fish-specific gene duplication. Genome Res. 2003;13:1067–81.PubMedCentralPubMedCrossRef
19.
Berghmans S, Murphey RD, Wienholds E, Neuberg D, Kutok JL, Fletcher CD, et al. tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors. Proc Natl Acad Sci U S A. 2005;102:407–12.PubMedCentralPubMedCrossRef
20.
Uhrbom L, Dai C, Celestino JC, Rosenblum MK, Fuller GN, Holland EC. Ink4a-Arf loss cooperates with KRas activation in astrocytes and neural progenitors to generate glioblastomas of various morphologies depending on activated Akt. Cancer Res. 2002;62:5551–8.PubMed
21.
22.
Kim SH, Speirs CK, Solnica-Krezel L, Ess KC. Zebrafish model of tuberous sclerosis complex reveals cell-autonomous and non-cell-autonomous functions of mutant tuberin. Dis Model Mech. 2011;4:255–67.PubMedCentralPubMedCrossRef
23.
Hawkins TA, Cavodeassi F, Erdelyi F, Szabo G, Lele Z. The small molecule Mek1/2 inhibitor U0126 disrupts the chordamesoderm to notochord transition in zebrafish. BMC Dev Biol. 2008;8:42.PubMedCentralPubMedCrossRef
24.
Restrepo A, Smith CA, Agnihotri S, Shekarforoush M, Kongkham PN, Seol HJ, et al. Epigenetic regulation of glial fibrillary acidic protein by DNA methylation in human malignant gliomas. Neuro Oncol. 2011;13:42–50.PubMedCentralPubMedCrossRef
25.
Friedmann-Morvinski D, Bushong EA, Ke E, Soda Y, Marumoto T, Singer O, et al. Dedifferentiation of neurons and astrocytes by oncogenes can induce gliomas in mice. Science. 2012;338:1080–4.PubMedCentralPubMedCrossRef
26.
Haigis KM, Kendall KR, Wang Y, Cheung A, Haigis MC, Glickman JN, et al. Differential effects of oncogenic K-Ras and N-Ras on proliferation, differentiation and tumor progression in the colon. Nat Genet. 2008;40:600–8.PubMedCentralPubMedCrossRef
27.
Song X, Andrew Allen R, Terence Dunn S, Fung KM, Farmer P, Gandhi S, et al. Glioblastoma with PNET-like components has a higher frequency of isocitrate dehydrogenase 1 (IDH1) mutation and likely a better prognosis than primary glioblastoma. Int J Clin Exp Pathol. 2011;4:651–60.PubMedCentralPubMed
28.
Cichowski K, Shih TS, Schmitt E, Santiago S, Reilly K, McLaughlin ME, et al. Mouse models of tumor development in neurofibromatosis type 1. Science. 1999;286:2172–6.PubMedCrossRef
29.
30.
Amsterdam A, Sadler KC, Lai K, Farrington S, Bronson RT, Lees JA, et al. Many ribosomal protein genes are cancer genes in zebrafish. PLoS Biol. 2004;2:E139.PubMedCentralPubMedCrossRef
31.
Sanai N, Alvarez-Buylla A, Berger MS. Neural stem cells and the origin of gliomas. N Engl J Med. 2005;353:811–22.PubMedCrossRef
32.
Marz M, Chapouton P, Diotel N, Vaillant C, Hesl B, Takamiya M, et al. Heterogeneity in progenitor cell subtypes in the ventricular zone of the zebrafish adult telencephalon. Glia. 2010;58:870–88.PubMed
33.
da Lee Y, Gianino SM, Gutmann DH. Innate neural stem cell heterogeneity determines the patterning of glioma formation in children. Cancer Cell. 2012;22:131–8.PubMedCentralCrossRef
34.
Shaw RJ, Cantley LC. Ras, PI(3)K and mTOR signalling controls tumour cell growth. Nature. 2006;441:424–30.PubMedCrossRef
35.
Nguyen AT, Emelyanov A, Koh CH, Spitsbergen JM, Parinov S, Gong Z. An inducible kras(V12) transgenic zebrafish model for liver tumorigenesis and chemical drug screening. Dis Model Mech. 2012;5:63–72.PubMedCentralPubMedCrossRef
36.
Roux PP, Shahbazian D, Vu H, Holz MK, Cohen MS, Taunton J, et al. RAS/ERK signaling promotes site-specific ribosomal protein S6 phosphorylation via RSK and stimulates cap-dependent translation. J Biol Chem. 2007;282:14056–64.PubMedCentralPubMedCrossRef
37.
Seok SH, Na YR, Han JH, Kim TH, Jung H, Lee BH, et al. Cre/loxP-regulated transgenic zebrafish model for neural progenitor-specific oncogenic Kras expression. Cancer Sci. 2010;101:149–54.PubMedCrossRef
38.
39.
White RM, Cech J, Ratanasirintrawoot S, Lin CY, Rahl PB, Burke CJ, et al. DHODH modulates transcriptional elongation in the neural crest and melanoma. Nature. 2011;471:518–22.PubMedCentralPubMedCrossRef
40.
Ridges S, Heaton WL, Joshi D, Choi H, Eiring A, Batchelor L, et al. Zebrafish screen identifies novel compound with selective toxicity against leukemia. Blood. 2012;119:5621–31.PubMedCentralPubMedCrossRef
41.
42.
Stern HM, Murphey RD, Shepard JL, Amatruda JF, Straub CT, Pfaff KL, et al. Small molecules that delay S phase suppress a zebrafish bmyb mutant. Nat Chem Biol. 2005;1:366–70.PubMedCrossRef
43.
Jeong JY, Kwon HB, Ahn JC, Kang D, Kwon SH, Park JA, et al. Functional and developmental analysis of the blood–brain barrier in zebrafish. Brain Res Bull. 2008;75:619–28.PubMedCrossRef
44.
Umans RA, Taylor MR. Zebrafish as a model to study drug transporters at the blood–brain barrier. Clin Pharmacol Ther. 2012;92:567–70.PubMedCrossRef
45.
Kwan KM, Fujimoto E, Grabher C, Mangum BD, Hardy ME, Campbell DS, et al. The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev Dyn. 2007;236:3088–99.PubMedCrossRef
46.
Urasaki A, Morvan G, Kawakami K. Functional dissection of the Tol2 transposable element identified the minimal cis-sequence and a highly repetitive sequence in the subterminal region essential for transposition. Genetics. 2006;174:639–49.PubMedCentralPubMedCrossRef
Metadata
Title
Oncogenic KRAS promotes malignant brain tumors in zebrafish
Authors
Bensheng Ju
Wenbiao Chen
Brent A Orr
Jan M Spitsbergen
Sujuan Jia
Christopher J Eden
Hannah E Henson
Michael R Taylor
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2015
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/s12943-015-0288-2

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