PUBLICATION

Development of zebrafish medulloblastoma-like PNET model by TALEN-mediated somatic gene inactivation

Authors: Shim, J., Choi, J.H., Park, M.H., Kim, H., Kim, J.H., Kim, S.Y., Hong, D., Kim, S., Lee, J.E., Kim, C.H., Lee, J.S., Bae, Y.K.
ID: ZDB-PUB-170915-11
Date: 2017
Source: Oncotarget 8: 55280-55297 (Journal)
Registered Authors: Bae, Young Ki, Choi, Jung-Hwa, Kim, Cheol-Hee, Lee, Jeong-Soo, Lee, Ji Eun, Park, Moon-Hak
Keywords: MPNST, PNET, TALEN, medulloblastoma, somatic inactivation
Datasets: GEO:GSE92260
MeSH Terms: none
PubMed: 28903419 Full text @ Oncotarget

Abstract

Genetically engineered animal tumor models have traditionally been generated by the gain of single or multiple oncogenes or the loss of tumor suppressor genes; however, the development of live animal models has been difficult given that cancer phenotypes are generally induced by somatic mutation rather than by germline genetic inactivation. In this study, we developed somatically mutated tumor models using TALEN-mediated somatic gene inactivation of cdkn2a/b or rb1 tumor suppressor genes in zebrafish. One-cell stage injection of cdkn2a/b-TALEN mRNA resulted in malignant peripheral nerve sheath tumors with high frequency (about 39%) and early onset (about 35 weeks of age) in F0 tp53^e7/e7 mutant zebrafish. Injection of rb1-TALEN mRNA also led to the formation of brain tumors at high frequency (58%, 31 weeks of age) in F0 tp53^e7/e7 mutant zebrafish. Analysis of each tumor induced by somatic inactivation showed that the targeted genes had bi-allelic mutations. Tumors induced by rb1 somatic inactivation were characterized as medulloblastoma-like primitive neuroectodermal tumors based on incidence location, histopathological features, and immunohistochemical tests. In addition, 3' mRNA Quanti-Seq analysis showed differential activation of genes involved in cell cycle, DNA replication, and protein synthesis; especially, genes involved in neuronal development were up-regulated.

Genes / Markers

Figures

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Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + TALEN4-rb1	standard conditions	Adult	whole organism decreased life span, abnormal	Fig. 3
rb1^zk3/zk3	standard conditions	Days 7-13	liver increased size, abnormal	Fig. S3
rb1^zk3/zk3	standard conditions	Days 7-13	swim bladder flattened, abnormal	Fig. S3
rb1^zk3/zk3	standard conditions	Days 7-13	whole organism decreased life span, abnormal	Fig. S3
tp53^zdf1/zdf1 + TALEN1-cdkn2a/b	standard conditions	Adult	eye anatomical region neoplastic, exacerbated	Fig. 1
tp53^zdf1/zdf1 + TALEN1-cdkn2a/b	standard conditions	Adult	eye neurofibrosarcoma increased amount, exacerbated	Fig. 1
tp53^zdf1/zdf1 + TALEN1-cdkn2a/b	standard conditions	Adult	whole organism decreased life span, abnormal	Fig. 1
tp53^zdf1/zdf1 + TALEN2-cdkn2a/b	standard conditions	Adult	eye anatomical region neoplastic, exacerbated	Fig. 1
tp53^zdf1/zdf1 + TALEN2-cdkn2a/b	standard conditions	Adult	eye neurofibrosarcoma increased amount, exacerbated	Fig. 1
tp53^zdf1/zdf1 + TALEN2-cdkn2a/b	standard conditions	Adult	whole organism decreased life span, abnormal	Fig. 1

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Mutations / Transgenics

Allele	Type	Affected Genomic Region
zdf1	Point Mutation	tp53
zk1	Small Deletion	cdkn2a/b
zk2	Small Deletion	cdkn2a/b
zk3	Small Deletion	rb1

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
malignant peripheral nerve sheath tumor	tp53^zdf1/zdf1 + TALEN1-cdkn2a/b	standard conditions	TAS
malignant peripheral nerve sheath tumor	tp53^zdf1/zdf1 + TALEN2-cdkn2a/b	standard conditions	TAS
medulloblastoma	tp53^zdf1/zdf1 + TALEN4-rb1	standard conditions	TAS

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Sequence Targeting Reagents

Target	Reagent	Reagent Type
cdkn2a/b	TALEN1-cdkn2a/b	TALEN
cdkn2a/b	TALEN2-cdkn2a/b	TALEN
rb1	TALEN4-rb1	TALEN

Fish

Fish
AB
AB + TALEN4-rb1
rb1^zk3/zk3
tp53^zdf1/zdf1
tp53^zdf1/zdf1 + TALEN1-cdkn2a/b
tp53^zdf1/zdf1 + TALEN2-cdkn2a/b
tp53^zdf1/zdf1 + TALEN4-rb1

Antibodies

Orthology

Gene	Orthology
cdkn2a/b

Engineered Foreign Genes

Mapping

Shim et al., 2017 ZDB-PUB-170915-11 PMID:28903419

Development of zebrafish medulloblastoma-like PNET model by TALEN-mediated somatic gene inactivation

Shim et al., 2017

ZDB-PUB-170915-11
PMID:28903419