PUBLICATION

Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development

Authors

Bresciani, E., Carrington, B., Yu, K., Kim, E.M.K., Zhen, T., Guzman, V.S., Broadbridge, E., Bishop, K., Kirby, M., Harper, U., Wincovitch, S., Dell'Orso, S., Sartorelli, V., Sood, R., Liu, P.P.

ID

ZDB-PUB-210908-15

Date

2021

Source

Blood advances 5(23): 4949-4962 (Journal)

Registered Authors

Bresciani, Erica, Sood, Raman

Keywords

none

Datasets

GEO:GSE158099, GEO:GSE158098, GEO:GSE169689, GEO:GSE158101

MeSH Terms

Hemangioblasts*
Zebrafish
Animals
Mice
Hematopoietic Stem Cells
Core Binding Factor Alpha 2 Subunit/genetics
Core Binding Factor Alpha 2 Subunit/metabolism*
GATA2 Transcription Factor/genetics
GATA2 Transcription Factor/metabolism*
Hematopoiesis/genetics
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*

(all 12)

PubMed

34492681 Full text @ Blood Adv

Abstract

RUNX1 is essential for the generation of hematopoietic stem cells (HSCs). Runx1 null mouse embryos lack definitive hematopoiesis and die in mid-gestation. However, even though zebrafish embryos with a runx1 W84X mutation have defects in early definitive hematopoiesis, some runx1W84X/W84X embryos can develop to fertile adults with blood cells of multi-lineages, raising the possibility that HSCs can emerge without RUNX1. Here, using three new zebrafish runx1-/- lines we uncovered the compensatory mechanism for runx1-independent hematopoiesis. We show that, in the absence of a functional runx1, a cd41-GFP+ population of hematopoietic precursors still emerge from the hemogenic endothelium and can colonize the hematopoietic tissues of the mutant embryos. Single-cell RNA sequencing of the cd41-GFP+ cells identified a set of runx1-/--specific signature genes during hematopoiesis. Significantly, gata2b, which normally acts upstream of runx1 for the generation of HSCs, was increased in the cd41-GFP+ cells in runx1- /- embryos. Interestingly, genetic inactivation of both gata2b and its paralog, gata2a, did not affect hematopoiesis. However, knocking out runx1 and any three of the four alleles of gata2a and gata2b abolished definitive hematopoiesis. Gata2 expression was also upregulated in hematopoietic cells in Runx1-/- mice, suggesting the compensatory mechanism is conserved. Our findings indicate that RUNX1 and GATA2 serve redundant roles for HSC production, acting as each other's safeguard.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
apln	runx1^hg96/hg96	standard conditions	Adult	kidney thrombocyte	RNA-Seq	Figure 1.
apln	runx1^hg97/hg97	standard conditions	Adult	kidney thrombocyte	RNA-Seq	Figure 1.
DsRed	runx1^hg96/hg96; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	Protruding-mouth	caudal hematopoietic tissue vasculature	IFL	Figure 2.
DsRed	runx1^hg96/hg96; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	14-19 somites	nucleate erythrocyte	IFL	Figure 1.
DsRed	runx1^hg96/hg96; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	Bud	nucleate erythrocyte	IFL	Figure 1.
DsRed	runx1^hg96/hg96; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	Long-pec	ventral wall of dorsal aorta vasculature	IFL	Figure 2.
DsRed	runx1^hg96/hg96; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	Protruding-mouth	ventral wall of dorsal aorta vasculature	IFL	Figure 2.
DsRed	runx1^hg96/+; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	Protruding-mouth	caudal hematopoietic tissue vasculature	IFL	Figure 2.
DsRed	runx1^hg96/+; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	14-19 somites	nucleate erythrocyte	IFL	Figure 1.
DsRed	runx1^hg96/+; la2Tg/la2Tg; sd2Tg/sd2Tg	standard conditions	Bud	nucleate erythrocyte	IFL	Figure 1.

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
en.gata2a.i4^hg123/+; gata2b^hg95/hg95; runx1^hg96/+	standard conditions	Days 45-89	head kidney hematopoietic multipotent progenitor cell normal amount, normal	Figure 6.
en.gata2a.i4^hg123/+; gata2b^hg95/hg95; runx1^hg96/hg96	standard conditions	Days 45-89	head kidney hematopoietic multipotent progenitor cell decreased amount, abnormal	Figure 6.
en.gata2a.i4^hg123/hg123	standard conditions	Days 30-44	head kidney hematopoietic multipotent progenitor cell normal amount, normal	Figure 6.
en.gata2a.i4^hg123/hg123; gata2b^hg95/+; runx1^hg96/hg96	standard conditions	Days 30-44	head kidney hematopoietic multipotent progenitor cell decreased amount, abnormal	Figure 6.
en.gata2a.i4^hg123/hg123; gata2b^hg95/hg95	standard conditions	Days 30-44	head kidney hematopoietic multipotent progenitor cell normal amount, normal	Figure 6.
en.gata2a.i4^hg123/hg123; gata2b^hg95/hg95; runx1^hg96/hg96	standard conditions	Days 30-44	head kidney hematopoietic multipotent progenitor cell decreased amount, abnormal	Figure 6.
runx1^hg96/+	standard conditions	Adult	whole organism morphology, normal	Figure 1.
runx1^hg96/+	standard conditions	Adult	whole organism viability, normal	Figure 1.
runx1^hg96/hg96	standard conditions	26+ somites	kidney cell morphology, normal	Figure 1.
runx1^hg96/hg96	standard conditions	26+ somites	nucleate erythrocyte cell morphology, normal	Figure 1.

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hg1		Point Mutation	runx1
hg94		Insertion	gata2b
hg95		Small Deletion	gata2b
hg96		Small Deletion	runx1
hg97		Small Deletion	runx1
hg98		Small Deletion	runx1
hg123		Small Deletion	en.gata2a.i4
la2Tg	Tg(-6.0itga2b:EGFP)	Transgenic Insertion
sd2Tg	Tg(gata1a:DsRed)	Transgenic Insertion
y171Tg	Tg(kdrl:mCherry-CAAX)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
en.gata2a.i4	CRISPR1-en.gata2a.i4	CRISPR
en.gata2a.i4	CRISPR3-en.gata2a.i4	CRISPR
gata2b	CRISPR1-gata2b	CRISPR
gata2b	CRISPR7-gata2b	CRISPR
runx1	CRISPR2-runx1	CRISPR
runx1	CRISPR3-runx1	CRISPR
runx1	TALEN1-runx1	TALEN

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Fish

Fish
en.gata2a.i4^hg123/hg123
en.gata2a.i4^hg123/hg123; gata2b^hg95/hg95
en.gata2a.i4^hg123/hg123; gata2b^hg95/hg95; runx1^hg96/hg96
en.gata2a.i4^hg123/hg123; gata2b^hg95/+; runx1^hg96/hg96
en.gata2a.i4^hg123/+; gata2b^hg95/hg95; runx1^hg96/hg96
en.gata2a.i4^hg123/+; gata2b^hg95/hg95; runx1^hg96/+
gata2b^hg95/hg95; runx1^hg96/hg96
runx1^hg96/hg96
runx1^hg96/hg96; gata2b^hg94/hg94
runx1^hg96/hg96; la2Tg/la2Tg; sd2Tg/sd2Tg

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed	EFG	DsRed
EGFP	EFG	EGFP
mCherry	EFG	mCherry

Mapping

Bresciani et al., 2021 ZDB-PUB-210908-15 PMID:34492681

Redundant mechanisms driven independently by RUNX1 and GATA2 for hematopoietic development

Bresciani et al., 2021

ZDB-PUB-210908-15
PMID:34492681