PUBLICATION

Stepwise crosstalk between aberrant Nf1, Tp53 and Rb signalling pathways induces gliomagenesis in zebrafish

Authors

Luo, J., Liu, P., Lu, C., Bian, W., Su, D., Zhu, C., Xie, S., Pan, Y., Li, N., Cui, W., Pei, D.S., Yang, X.

ID

ZDB-PUB-201208-50

Date

2020

Source

Brain : a journal of neurology 144(2): 615-635 (Journal)

Registered Authors

Keywords

glioma, nf1, rb1, tp53, zebrafish

Datasets

GEO:GSE129899

MeSH Terms

Signal Transduction*
Tumor Suppressor Protein p53/metabolism
Female
Brain Neoplasms/metabolism*
Brain Neoplasms/pathology
Zebrafish Proteins/metabolism
Male
Glioma/metabolism*
Glioma/pathology
Retinoblastoma Protein/metabolism
Neurofibromatosis 1/metabolism
Animals
Mutation
Animals, Genetically Modified
Zebrafish

(all 15)

PubMed

33279959 Full text @ Brain

Abstract

The molecular pathogenesis of glioblastoma indicates that RTK/Ras/PI3K, RB and TP53 pathways are critical for human gliomagenesis. Here, several transgenic zebrafish lines with single or multiple deletions of nf1, tp53 and rb1 in astrocytes, were established to genetically induce gliomagenesis in zebrafish. In the mutant with a single deletion, we found only the nf1 mutation low-efficiently induced tumour incidence, suggesting that the Nf1 pathway is critical for the initiation of gliomagenesis in zebrafish. Combination of mutations, nf1;tp53 and rb1;tp53 combined knockout fish, showed much higher tumour incidences, high-grade histology, increased invasiveness, and shortened survival time. Further bioinformatics analyses demonstrated the alterations in RTK/Ras/PI3K, cell cycle, and focal adhesion pathways, induced by abrogated nf1, tp53, or rb1, were probably the critical stepwise biological events for the initiation and development of gliomagenesis in zebrafish. Gene expression profiling and histological analyses showed the tumours derived from zebrafish have significant similarities to the subgroups of human gliomas. Furthermore, temozolomide treatment effectively suppressed gliomagenesis in these glioma zebrafish models, and the histological responses in temozolomide-treated zebrafish were similar to those observed in clinically treated glioma patients. Thus, our findings will offer a potential tool for genetically investigating gliomagenesis and screening potential targeted anti-tumour compounds for glioma treatment.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
akt1	AB	standard conditions	Adult	brain	RTPCR	Fig. 6
akt1	AB	standard conditions	Adult	brain	WB	Fig. 6
akt1	nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Adult	brain neoplasm	WB	Fig. 6
akt1	nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Adult	brain	RTPCR	Fig. 6
akt1	nf1a^{zf3453a/zf3453a}; zf3453Tg (AB)	standard conditions	Adult	brain	RTPCR	Fig. 6
akt1	rb1^{zf3454a/zf3454a}; tp53^{zf3455a/zf3455a}; zf3454Tg; zf3455Tg	standard conditions	Adult	brain	RTPCR	Fig. 6
casp3a	AB	standard conditions	Adult	brain	RTPCR	Fig. 6
casp3a	nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Adult	brain	RTPCR	Fig. 6
casp3a	nf1a^{zf3453a/zf3453a}; zf3453Tg (AB)	standard conditions	Adult	brain	RTPCR	Fig. 6
casp3a	rb1^{zf3454a/zf3454a}; tp53^{zf3455a/zf3455a}; zf3454Tg; zf3455Tg	standard conditions	Adult	brain	RTPCR	Fig. 6

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
nf1a^{zf3453a/zf3453a}; rb1^{zf3454a/zf3454a}; zf3453Tg; zf3454Tg	standard conditions	Adult	brain glioblastoma present, abnormal	Fig. 5
nf1a^{zf3453a/zf3453a}; rb1^{zf3454a/zf3454a}; zf3453Tg; zf3454Tg	standard conditions	Adult	whole organism viability, abnormal	Fig. 5
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Days 45-89	brain ctnnb1 expression increased amount, abnormal	Fig. 4
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Days 45-89	brain gfap expression increased amount, abnormal	Fig. 4
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Days 45-89	brain pcna expression increased amount, abnormal	Fig. 4
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Days 45-89 to Adult	brain glioblastoma present, abnormal	Fig. 4
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Adult	brain akt1 expression increased amount, abnormal	Fig. 6
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Adult	brain casp3a expression decreased amount, abnormal	Fig. 6
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Adult	brain casp3b expression decreased amount, abnormal	Fig. 6
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	Adult	brain ccnd1 expression increased amount, abnormal	Fig. 8

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
mi2001Tg	Tg(gfap:GFP)	Transgenic Insertion
zf3453a		Unknown	nf1a
zf3453Tg	Tg(gfap:Cas9-2A-mCherry,rnu6-z2:CRISPR3-nf1a)	Transgenic Insertion
zf3454a		Unknown	rb1
zf3454Tg	Tg(gfap:Cas9-2A-mCherry,rnu6-z2:CRISPR6-rb1)	Transgenic Insertion
zf3455a		Unknown	tp53
zf3455Tg	Tg(gfap:Cas9-2A-mCherry,rnu6-z2:CRISPR12-tp53)	Transgenic Insertion

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
brain glioblastoma multiforme	nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg	standard conditions	TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
nf1a	CRISPR3-nf1a	CRISPR
nf1b	CRISPR2-nf1b	CRISPR
rb1	CRISPR6-rb1	CRISPR
tp53	CRISPR12-tp53	CRISPR

Fish

Fish
AB
AB/C32
nf1a^{zf3453a/zf3453a}; rb1^{zf3454a/zf3454a}; zf3453Tg; zf3454Tg
nf1a^{zf3453a/zf3453a}; tp53^{zf3455a/zf3455a}; zf3453Tg; zf3455Tg
nf1a^{zf3453a/zf3453a}; zf3453Tg (AB)
rb1^{zf3454a/zf3454a}; tp53^{zf3455a/zf3455a}; zf3454Tg; zf3455Tg
rb1^{zf3454a/zf3454a}; zf3454Tg (AB)
tp53^{zf3455a/zf3455a}; zf3455Tg (AB)
WT

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Cas9	EFG	Cas9
GFP	EFG	GFP
mCherry	EFG	mCherry

Mapping

Luo et al., 2020 ZDB-PUB-201208-50 PMID:33279959

Stepwise crosstalk between aberrant Nf1, Tp53 and Rb signalling pathways induces gliomagenesis in zebrafish

Luo et al., 2020

ZDB-PUB-201208-50
PMID:33279959