PUBLICATION

A zebrafish model of Poikiloderma with Neutropenia recapitulates the human syndrome hallmarks and traces back neutropenia to the myeloid progenitor

Authors

Colombo, E.A., Carra, S., Fontana, L., Bresciani, E., Cotelli, F., Larizza, L.

ID

ZDB-PUB-151103-3

Date

2015

Source

Scientific Reports 5: 15814 (Journal)

Registered Authors

Bresciani, Erica, Cotelli, Franco

Keywords

Disease model, Diseases, Medical genetics

MeSH Terms

Skin Diseases/genetics
Skin Diseases/metabolism
Stem Cells/metabolism*
Humans
Zebrafish/genetics*
Myeloid Cells/metabolism*
Neutropenia/genetics*
Down-Regulation/genetics
Skin Abnormalities/genetics*
Morpholinos/genetics
Phenotype
RNA Splicing/genetics
RNA, Small Nuclear/genetics
Animals

(all 14)

PubMed

26522474 Full text @ Sci. Rep.

Abstract

Poikiloderma with Neutropenia (PN) is an autosomal recessive genodermatosis characterized by early-onset poikiloderma, pachyonychia, hyperkeratosis, bone anomalies and neutropenia, predisposing to myelodysplasia. The causative C16orf57/USB1 gene encodes a conserved phosphodiesterase that regulates the stability of spliceosomal U6-RNA. The involvement of USB1 in splicing has not yet allowed to unveil the pathogenesis of PN and how the gene defects impact on skin and bone tissues besides than on the haematological compartment. We established a zebrafish model of PN using a morpholino-knockdown approach with two different splicing morpholinos. Both usb1-depleted embryos displayed developmental abnormalities recapitulating the signs of the human syndrome. Besides the pigmentation and osteochondral defects, usb1-knockdown caused defects in circulation, manifested by a reduced number of circulating cells. The overall morphant phenotype was also obtained by co-injecting sub-phenotypic dosages of the two morpholinos and could be rescued by human USB1 RNA. Integrated in situ and real-time expression analyses of stage-specific markers highlighted defects of primitive haematopoiesis and traced back the dramatic reduction in neutrophil myeloperoxidase to the myeloid progenitors showing down-regulated pu.1 expression. Our vertebrate model of PN demonstrates the intrinsic requirement of usb1 in haematopoiesis and highlights PN as a disorder of myeloid progenitors associated with bone marrow dysfunction.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
DsRed	s843Tg; sd2Tg	control	Long-pec	nucleate erythrocyte	IFL	Fig. 4
DsRed	s843Tg; sd2Tg + MO3-usb1	control	Long-pec	nucleate erythrocyte	IFL	Fig. 4
EGFP	s843Tg; sd2Tg	control	Long-pec	vasculature	IFL	Fig. 4
EGFP	s843Tg; sd2Tg + MO3-usb1	control	Long-pec	vasculature	IFL	Fig. 4
gata1a	AB	control	Prim-5	erythroid lineage cell	ISH	Fig. 5
gata1a	AB + MO3-usb1	control	Prim-5	erythroid lineage cell	ISH	Fig. 5
GFP	i114Tg; nz50Tg	control	Long-pec	neutrophil	IFL	Fig. 4
GFP	i114Tg; nz50Tg	control	Long-pec	neutrophil	IFL	Fig. 6
GFP	i114Tg; nz50Tg + MO3-usb1	control	Long-pec	neutrophil	IFL	Fig. 6
GFP	i114Tg; nz50Tg + MO3-usb1	control	Long-pec	neutrophil	IFL	Fig. 4

1 - 10 of 52

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO3-usb1	control	26+ somites	neutrophil mpx expression decreased amount, abnormal	Fig. 5
AB + MO3-usb1	control	26+ somites	neutrophil mpx expression decreased distribution, abnormal	Fig. 5
AB + MO3-usb1	control	26+ somites	neutrophil decreased amount, abnormal	Fig. 5
AB + MO3-usb1	control	26+ somites	whole organism mpx expression decreased amount, abnormal	Fig. 5
AB + MO3-usb1	control	Prim-15	neutrophil mpx expression decreased amount, abnormal	Fig. 5
AB + MO3-usb1	control	Prim-15	neutrophil mpx expression decreased distribution, abnormal	Fig. 5
AB + MO3-usb1	control	Prim-15	neutrophil decreased amount, abnormal	Fig. 5
AB + MO3-usb1	control	Prim-15	whole organism mpx expression decreased amount, abnormal	Fig. 5
AB + MO3-usb1	control	Long-pec	nucleate erythrocyte decreased amount, abnormal	Fig. 5
AB + MO3-usb1	control	Day 5	ceratohyal cartilage displaced, abnormal	Fig. 4

1 - 10 of 49

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Mutations / Transgenics

Allele	Construct	Type
i114Tg	Tg(mpx:GFP)	Transgenic Insertion
nz50Tg	Tg(lyz:DsRed2)	Transgenic Insertion
s843Tg	Tg(kdrl:EGFP)	Transgenic Insertion
sd2Tg	Tg(gata1a:DsRed)	Transgenic Insertion

1 - 4 of 4

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
ablepharon macrostomia syndrome	AB + MO4-usb1	control	TAS
poikiloderma with neutropenia	AB + MO3-usb1	control	TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
tp53	MO4-tp53	MRPHLNO
usb1	MO3-usb1	MRPHLNO
usb1	MO4-usb1	MRPHLNO

Fish

Fish
AB
AB + MO3-usb1
AB + MO3-usb1 + MO4-usb1
AB + MO4-usb1
i114Tg; nz50Tg
i114Tg; nz50Tg + MO3-usb1
s843Tg; sd2Tg
s843Tg; sd2Tg + MO3-usb1

Antibodies

Orthology

Gene	Orthology
usb1

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed	EFG	DsRed
DsRed2	EFG	DsRed2
EGFP	EFG	EGFP
GFP	EFG	GFP

Mapping

Colombo et al., 2015 ZDB-PUB-151103-3 PMID:26522474

A zebrafish model of Poikiloderma with Neutropenia recapitulates the human syndrome hallmarks and traces back neutropenia to the myeloid progenitor

Colombo et al., 2015

ZDB-PUB-151103-3
PMID:26522474