PUBLICATION

First quantitative high-throughput screen in zebrafish identifies novel pathways for increasing pancreatic β-cell mass

Authors

Wang, G., Rajpurohit, S.K., Delaspre, F., Walker, S.L., White, D.T., Ceasrine, A., Kuruvilla, R., Li, R.J., Shim, J.S., Liu, J.O., Parsons, M.J., Mumm, J.S.

ID

ZDB-PUB-150729-3

Date

2015

Source

eLIFE 4: (Journal)

Registered Authors

Delaspre, Fabien, Mumm, Jeff, Rajpurohit, Surendra, Walker, Steven, Wang, Guangliang (Johnny)

Keywords

NF-κB, beta cell, cell biology, developmental biology, diabetes, high-throughput screening, mouse, serotonin, stem cells, whole-organism drug discovery, zebrafish

MeSH Terms

Cell Differentiation/drug effects*
Automation, Laboratory/methods
High-Throughput Screening Assays/methods*
Animals
Zebrafish/physiology*
Drug Discovery/methods*
Insulin-Secreting Cells/drug effects
Insulin-Secreting Cells/physiology*
Cell Proliferation/drug effects*

(all 9)

PubMed

26218223 Full text @ Elife

Abstract

Whole-organism chemical screening can circumvent bottlenecks that impede drug discovery. However, in vivo screens have not attained throughput capacities possible with in vitro assays. We therefore developed a method enabling in vivo high-throughput screening (HTS) in zebrafish, termed automated reporter quantification in vivo (ARQiv). Here, ARQiv was combined with robotics to fully actualize whole-organism HTS (ARQiv-HTS). In a primary screen, this platform quantified cell-specific fluorescent reporters in >500,000 transgenic zebrafish larvae to identify FDA-approved drugs that increased the number of insulin-producing β cells in the pancreas. Twenty-four drugs were confirmed as inducers of endocrine differentiation and/or stimulators of β-cell proliferation. Further, we discovered novel roles for NF-κB signaling in regulating endocrine differentiation and for serotonergic signaling in selectively stimulating β-cell proliferation. These studies demonstrate the power of ARQiv-HTS for drug discovery and provide unique insights into signaling pathways controlling β-cell mass, potential therapeutic targets for treating diabetes.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
EGFP	jh10Tg	chemical treatment: parthenolide	Day 5	pancreatic B cell nucleus	IHC	Fig. 6
EGFP	jh10Tg	control	Day 5	pancreatic B cell nucleus	IHC	Fig. 6
EGFP	jh10Tg	chemical treatment: RO4929097	Day 5	pancreatic B cell nucleus	IHC	Fig. 6
EGFP	jh10Tg; lmc01Tg	chemical treatment: parthenolide	Day 5	pancreatic B cell nucleus	IHC	Fig. 4
EGFP	jh10Tg; lmc01Tg	control	Day 5	pancreatic B cell nucleus	IHC	Fig. 4
EGFP	jh10Tg; lmc01Tg	chemical treatment: RO4929097	Day 5	pancreatic B cell nucleus	IHC	Fig. 4
EGFP	jh11Tg; nc1Tg	control	Day 5	pancreas endocrine cell	IFL	Fig. 5
EGFP	nl1Tg	chemical treatment: parthenolide	Day 5	pancreas endocrine cell	IFL	Fig. 3
EGFP	nl1Tg	control	Day 5	pancreas endocrine cell	IFL	Fig. 3
mCherry	jh11Tg; nc1Tg	control	Day 5	pancreas endocrine cell	IFL	Fig. 5

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: parthenolide	Adult	pancreatic B cell cell population proliferation increased process quality, abnormal	Fig. 6
jh10Tg	chemical treatment: fluoxetine	Day 5	pancreatic B cell increased amount, abnormal	Fig. 6
jh10Tg	chemical treatment: parthenolide	Day 5	pancreatic B cell cell population proliferation increased process quality, abnormal	Fig. 6
jh10Tg	chemical treatment: parthenolide	Day 5	pancreatic B cell increased amount, abnormal	Fig. 6
jh10Tg	chemical treatment: RO4929097	Day 5	pancreatic B cell increased amount, abnormal	Fig. 6
jh10Tg	chemical treatment: serotonin	Day 5	pancreatic B cell increased amount, abnormal	Fig. 6
jh10Tg; lmc01Tg	chemical treatment: parthenolide	Day 5	pancreatic B cell increased amount, abnormal	Fig. 4
jh10Tg; lmc01Tg	chemical treatment: RO4929097	Day 5	pancreatic B cell increased amount, abnormal	Fig. 4
jh10Tg; lmc01Tg	chemical treatment: RO4929097	Day 5	pancreatic D cell increased amount, abnormal	Fig. 4
lmc01Tg	chemical treatment: parthenolide	Day 5	pancreatic B cell increased amount, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type
jh10Tg	Tg(ins:hmgb1-EGFP)	Transgenic Insertion
jh11Tg	Tg(EPV.Tp1-Ocu.Hbb2:hmgb1-mCherry)	Transgenic Insertion
lmc01Tg	Tg(ins:PhiYFP-NTR,sst2:TagRFP)	Transgenic Insertion
nc1Tg	Tg(6xHsa.NFKB:EGFP)	Transgenic Insertion
nl1Tg	TgBAC(neurod1:EGFP)	Transgenic Insertion
ulg515Tg	Tg(pax6b:GFP)	Transgenic Insertion

1 - 6 of 6

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
type 1 diabetes mellitus			TAS

Sequence Targeting Reagents

Fish

Fish
jh10Tg
jh10Tg; lmc01Tg
jh11Tg; nc1Tg
lmc01Tg
nl1Tg
WT

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab2-serotonin	polyclonal		IgG	Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GFP	EFG	GFP
mCherry	EFG	mCherry
NTR	EFG	NTR
PhiYFP	EFG	PhiYFP
RFP	EFG	RFP
TagRFP	EFG	TagRFP

Mapping

Wang et al., 2015 ZDB-PUB-150729-3 PMID:26218223

First quantitative high-throughput screen in zebrafish identifies novel pathways for increasing pancreatic β-cell mass

Wang et al., 2015

ZDB-PUB-150729-3
PMID:26218223