PUBLICATION

Ccdc103 promotes myeloid cell proliferation and migration independent of motile cilia

Authors

Falkenberg, L.G., Beckman, S.A., Ravisankar, P., Dohn, T.E., Waxman, J.S.

ID

ZDB-PUB-210525-14

Date

2021

Source

Disease models & mechanisms 14(5): (Journal)

Registered Authors

Waxman, Joshua

Keywords

CCDC103, Cell migration, Microtubules, Myeloid cells, Primary ciliary dyskinesia, Zebrafish

MeSH Terms

Cell Proliferation
Microtubules/metabolism
Zebrafish/embryology
Green Fluorescent Proteins/metabolism
HL-60 Cells
Stem Cells/metabolism
Animals
Neutrophils/metabolism
Cilia/metabolism*
Myeloid Cells/cytology*
Myeloid Cells/metabolism*
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Cell Movement*
Humans
Embryo, Nonmammalian/metabolism
Protein Binding
Mutation/genetics
HEK293 Cells

(all 19)

PubMed

34028558 Full text @ Dis. Model. Mech.

Abstract

The pathology of primary ciliary dyskinesia (PCD) is predominantly attributed to impairment of motile cilia. However, PCD patients also have perplexing functional defects in myeloid cells, which lack motile cilia. Here, we show that coiled-coil domain-containing protein 103 (CCDC103), one of the genes that, when mutated, is known to cause PCD, is required for the proliferation and directed migration of myeloid cells. CCDC103 is expressed in human myeloid cells, where it colocalizes with cytoplasmic microtubules. Zebrafish ccdc103/schmalhans (smh) mutants have macrophages and neutrophils with reduced proliferation, abnormally rounded cell morphology and an inability to migrate efficiently to the site of sterile wounds, all of which are consistent with a loss of cytoplasmic microtubule stability. Furthermore, we demonstrate that direct interactions between CCDC103 and sperm associated antigen 6 (SPAG6), which also promotes microtubule stability, are abrogated by CCDC103 mutations from PCD patients, and that spag6 zebrafish mutants recapitulate the myeloid defects observed in smh mutants. In summary, we have illuminated a mechanism, independent of motile cilia, to explain functional defects in myeloid cells from PCD patients. This article has an associated First Person interview with the first author of the paper.

Genes / Markers

Figures

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Expression

Gene	Antibody	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
dnaaf19	Ab2-ccdc103	dnaaf19^{tn222a/tn222a}; gl21Tg	standard conditions	20-25 somites	Not Detected	myeloid cell	IHC	Fig. S3
dnaaf19	Ab2-ccdc103	gl21Tg	standard conditions	20-25 somites		myeloid cell	IHC	Fig. S2
dnaaf19	Ab2-ccdc103	gl21Tg	standard conditions	20-25 somites		myeloid cell	IHC	Fig. S3
dnaaf19	Ab2-ccdc103	uwm1Tg	standard conditions	Prim-5		neutrophil perinuclear region of cytoplasm	IHC	Fig. 1.
dnaaf19		WT	standard conditions	14-19 somites		anterior lateral plate mesoderm myeloid lineage restricted progenitor cell	ISH	Fig. 1.
dnaaf19		WT	standard conditions	14-19 somites		pronephros	ISH	Fig. 1.
EGFP		gl21Tg	standard conditions	20-25 somites		myeloid cell	IHC	Fig. S2
GFP		uwm1Tg	standard conditions	Prim-5		neutrophil	IHC	Fig. 1.

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
dnaaf19^{tn222a/tn222a}	standard conditions	Prim-5	macrophage decreased amount, abnormal	Fig. S5
dnaaf19^{tn222a/tn222a}	standard conditions	Prim-5	neutrophil decreased amount, abnormal	Fig. S5
dnaaf19^{tn222a/tn222a}; gl21Tg	standard conditions	20-25 somites	myeloid cell dnaaf19 expression absent, abnormal	Fig. S3
dnaaf19^{tn222a/tn222a}; gl21Tg	standard conditions	Prim-5	myeloid lineage restricted progenitor cell decreased amount, abnormal	Fig. 3.
dnaaf19^{tn222a/tn222a}; uwm1Tg	standard conditions	Prim-5	myeloid cell cell population proliferation decreased occurrence, abnormal	Fig. 4.
dnaaf19^{tn222a/tn222a}; uwm1Tg	standard conditions	Prim-5	neutrophil decreased amount, abnormal	Fig. 3.
dnaaf19^{tn222a/tn222a}; uwm1Tg	light damage: yolk, chemical treatment by environment: paclitaxel	Prim-5	myeloid cell cell population proliferation occurrence, ameliorated	Fig. 6.
dnaaf19^{tn222a/tn222a}; uwm1Tg	light damage: yolk, chemical treatment by environment: paclitaxel	Prim-5	neutrophil migration process efficacy, ameliorated	Fig. 6.
dnaaf19^{tn222a/tn222a}; uwm1Tg	light damage: yolk, chemical treatment by environment: paclitaxel	Prim-5	neutrophil shape, ameliorated	Fig. 6.
dnaaf19^{tn222a/tn222a}; uwm1Tg	light damage: yolk	Prim-5	myeloid cell cell population proliferation decreased occurrence, abnormal	Fig. 6.

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
ci1013		Small Deletion	spag6
gl21Tg	Tg(-5.3spi1b:EGFP)	Transgenic Insertion
tn222a		Point Mutation	dnaaf19
uwm1Tg	Tg(mpx:GFP)	Transgenic Insertion
w200Tg	Tg(mpeg1:YFP)	Transgenic Insertion

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
primary ciliary dyskinesia	dnaaf19^{tn222a/tn222a}	standard conditions	TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
spag6	CRISPR1-spag6	CRISPR
spag6	CRISPR2-spag6	CRISPR

Fish

Fish
dnaaf19^{tn222a/tn222a}
dnaaf19^{tn222a/tn222a}; gl21Tg
dnaaf19^{tn222a/tn222a}; uwm1Tg
dnaaf19^{tn222a/tn222a}; w200Tg
gl21Tg
spag6^{ci1013/ci1013}
spag6^{ci1013/ci1013}
spag6^{ci1013/ci1013}; gl21Tg
spag6^{ci1013/ci1013}; uwm1Tg
uwm1Tg

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab2-ccdc103	polyclonal	dnaaf19		Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GFP	EFG	GFP
YFP	EFG	YFP

Mapping

Falkenberg et al., 2021 ZDB-PUB-210525-14 PMID:34028558

Ccdc103 promotes myeloid cell proliferation and migration independent of motile cilia

Falkenberg et al., 2021

ZDB-PUB-210525-14
PMID:34028558