Table S1. Animal models of CCM
Gene
Organism
Details
heart of
glass
(HEG1)
Zebrafish
mutant allele/
morpholino
heart of
glass
(HEG1)
Zebrafish
santa
(KRIT1)
Phenotype
Reference
Embryonic lethal with massively dilated hearts unable to
circulate blood. Only a single cell layer in the myocardial
wall, normal myocardial and endocardial cell number.
(Mably et al.,
2003)
morpholino
Dilated heart chambers but normal vessel patterning.
Blocked circulation due to a failure in vessel lumen
formation.
(Kleaveland
et al., 2009)
Zebrafish
morpholino
Embryonic lethal with massively dilated hearts unable to
circulate blood. Only a single cell layer in the myocardial
wall, normal myocardial and endocardial cell number.
(Mably et al.,
2006)
santa
(KRIT1)
Zebrafish
mutant allele
Normal vascular patterning but vascular morphogenesis
defects, failed circulation initiation, dilation of major
vessels and heart. Normal cell number and cell-cell
junctions, progressive dilation due to cell spreading.
(Hogan et al.,
2008)
rap1b;
santa
(RAP1B;
KRIT1)
valentine
(CCM2)
Zebrafish
morpholino
Co-injection of low doses of santa and rap1b morpholinos
led to synergistic enhancement of the dilated heart
phenotype
(Gore et al.,
2008)
Zebrafish
mutant allele/
morpholino
Embryonic lethal with massively dilated hearts unable to
circulate blood. Only a single cell layer in the myocardial
wall, normal myocardial and endocardial cell number.
(Mably et al.,
2006)
valentine
(CCM2)
Zebrafish
mutant allele/
morpholino
Normal vascular patterning but vascular morphogenesis
defects, failed circulation initiation, dilation of major
vessels and heart. Normal cell number and cell-cell
junctions, progressive dilation due to cell spreading.
(Hogan et al.,
2008)
valentine
(CCM2)
Zebrafish
morpholino
Dilated heart chambers but normal vessel patterning.
Blocked circulation due to a failure in vessel lumen
formation.
(Kleaveland
et al., 2009)
santa/
valentine;
heart of
glass
(KRIT1/
CCM2;
HEG1)
santa;
valentine
(KRIT1;
CCM2)
Zebrafish
comorpholino
Non-phenotypic doses of santa or valentine morpholinos
co-injected with non-phenotypic doses of heart of glass
morpholinos leads to dilated heart phenotype.
(Mably et al.,
2006)
Zebrafish
double
mutant
Phenotype is identical to single mutants. Normal vascular
patterning but vascular morphogenesis defects. Failed
circulation initiation and severe and progressive dilation of
(Hogan et al.,
2008)
major vessels and heart.
ccm3a/b
(PDCD10)
Zebrafish
mutant allele/
morpholino
Progressive dilation of major vessels and heart.
Indistinguishable from santa and valentine phenotypes.
(Voss et al.,
2009)
ccm3a/b
(PDCD10)
Zebrafish
morpholino
Embryonic lethal from dilated heart phenotype, thin
myocardium and blocked circulation.
(Zheng et al.,
2010)
ccm3a/b
(PDCD10)
Zebrafish
morpholino
Gross dilation and mispatterning of cranial vessels, mostly
not lumenized. Lack of circulation and pericardial edema,
but no dilated heart. Normal endothelial cell numbers but
increases in cell membrane protrusion number and size.
(Yoruk et al.,
2012)
stk25a
(STK25)
Zebrafish
morpholino
Misconnections in cranial vasculature.
(Yoruk et al.,
2012)
stk25b
(STK25)
Zebrafish
morpholino
Enlarged cranial vasculature. Pericardial and venous plexus
edemas, no enlarged heart and normal blood circulation.
(Yoruk et al.,
2012)
stk24;
stk25a/b
(STK24;
STK25)
ccm3a/b;
stk24/
stk25a/b
(PDCD10,
STK24;
STK25)
ccm3a/b;
stk25a/b
(PDCD10;
STK25)
valentine;
ccm3a/b
(CCM2;
PDCD10)
ccm2l
(CCM2L)
Zebrafish
comorpholino
Morpholinos for stk24, stk25a and stk25b all co-injected
led to the dilated heart phenotype.
(Zheng et al.,
2010)
Zebrafish
comorpholino
Morpholino targeting ccm3a/b co-injected with either
stk24, stk25a or stk25b morpholinos led to the dilated
heart phenotype.
(Zheng et al.,
2010)
Zebrafish
comorpholino
Non-phenotypic doses of ccm3 morpholinos co-injected
with either stk25a or stk25b synergized to lead to cranial
dilations.
(Yoruk et al.,
2012)
Zebrafish
morpholino
Non-phenotypic doses of ccm3 morpholinos injected into
valentine heterozygote led to cranial vasculature defects
more severe than the complete loss of ccm3.
(Yoruk et al.,
2012)
Zebrafish
morpholino
Dilation of atrium and inflow tract; occasional cessation of
blood circulation. SIVs have unusual growth patterns, some
ISVs fail to lumenize, likely due to reduced blood
circulation.
(Rosen et al.,
2013)
Pdcd10
(PDCD10)
Drosophila
dsRNA
Tracheal tubes branch normally, but fail to lumenize.
(Chan et al.,
2011)
Wheezy
(GCKIII)
Drosophila
dsRNA
Tracheal tubes branch normally, but fail to lumenize.
(Chan et al.,
2011)
Ccm1
Mouse
Global
Failed lumen in first branchial arch artery and abnormal
(Whitehead
(KRIT1)
Knockout
yolk sac vascular remodeling leading to failed circulation at
E8.5-E9.5
et al., 2004)
Ccm1+/(KRIT1)
Mouse
Heterozygous
Increased basal vascular leak in brain and lungs, increased
LPS-induced leaks in brain and pulmonary vessels.
(Stockton et
al., 2010)
Ccm1
(KRIT1)
Mouse
Endothelial
Inducible
Knockout;
Cdh5-CreERT2
Induced of Cre at P3 led to CCM-like lesions in the
cerebellum associated with EndMT-like traits at P13.
(Maddaluno
et al., 2013)
Ccm1+/-;
p53+/(KRIT1,
P53)
Ccm1+/-;
Msh2-/(KRIT1;
MSH2)
Ccm2+/(CCM2)
Mouse
Sensitized
Heterozygous
~55% mice had CCM-like vascular lesions
(Plummer et
al., 2004)
Mouse
Sensitized
Heterozygous
~50% of mice had CCM-like lesions.
(McDonald et
al., 2011)
Mouse
Heterozygous
Very low rate of brain vascular malformations.
(Plummer et
al., 2006)
Ccm2+/(CCM2)
Mouse
Heterozygous
Increased basal vascular leak in brain and lungs, increased
LPS-induced leaks in brain and pulmonary vessels.
(Stockton et
al., 2010)
Ccm2+/(CCM2)
Mouse
Gene Trap
Heterozygous,
Endothelial
Heterozygous
No overt phenotype, increased vascular permeability to
Evans blue after VEGF treatment.
(Whitehead
et al., 2009),
Ccm2
(CCM2)
Mouse
Global
Knockout
Embryonic lethal by E10.5. No phenotypic difference at
E8.5, but pale yolk sacs and pericardial edema at E9.5.
(Boulday et
al., 2009)
Ccm2
(CCM2)
Mouse
Global
Knockout;
HPRT-Cre
Normal vasculogenesis/patterning and heart development
at E8.0. Failed lumen in first branchial arch artery and
abnormal yolk sac vascular remodeling leading to failed
circulation at E8.5-E9.5; dead by E10.
(Whitehead
et al., 2009)
Ccm2
(CCM2)
Mouse
Global
Knockout
No remodeled vessels within yolk sac at E9.5; angiogenic
arrest during cardiovascular development. Does not recruit
vascular smooth muscle cells. No aortic lumen. Defective
heart morphogenesis.
(Cunningham
et al., 2011)
Ccm2
(CCM2)
Mouse
Neural
Knockout;
Nestin-Cre
No overt phenotype
(Boulday et
al., 2009)
Ccm2
(CCM2)
Mouse
Neural
Knockout;
No overt phenotype
(Whitehead
Nestin-Cre
et al., 2009)
Ccm2
(CCM2)
Mouse
Neural
Knockout;
Nestin-Cre
No overt phenotype.
(Chan et al.,
2011)
Ccm2
(CCM2)
Mouse
Smooth
Muscle
Knockout;
Transgelin-Cre
No overt phenotype
(Whitehead
et al., 2009)
Ccm2
(CCM2)
Mouse
Endothelial
Knockout;
Tie2-Cre
Embryonic lethal by E11.5. Pericardial edema at E10.5 with
signs of hemorrhage in pericardial cavity and trunk. Normal
vasculogenesis but impaired angiogenesis and impaired
recruitment of vascular smooth muscle cells.
(Boulday et
al., 2009)
Ccm2
(CCM2)
Mouse
Endothelial
Knockout;
Tie2-Cre
Normal vasculogenesis/patterning and heart development
at E8.0. Failed lumen in first branchial arch artery and
abnormal yolk sac vascular remodeling leading to failed
circulation at E8.5-E9.5; dead by E10.
(Whitehead
et al., 2009)
Ccm2
(CCM2)
Mouse
Endothelial
Inducible
Knockout;
Cdh5-CreERT2
Induction of Cre at P1- 100% of mice have CCM-like lesions
in the cerebellum and retina by P8. Induction of Cre at P21no lesions at 2 months.
(Boulday et
al., 2011)
Ccm2
(CCM2)
Mouse
Endothelial
Knockout;
Tie2-Cre
No remodeled vessels within yolk sac at E9.5; angiogenic
arrest during cardiovascular development. Does not recruit
vascular smooth muscle cells. No aortic lumen. Defective
heart morphogenesis.
(Cunningham
et al., 2011)
Ccm2
(CCM2)
Mouse
Inducible
Knockout;
PDGFb-iCreER
Induction at P1 expresses Cre throughout the brain
vasculature. CCM-like lesions no earlier than 2-months of
age. Fully penetrant at 6-months.
(Chan et al.,
2011)
Ccm2
(CCM2)
Mouse
Inducible
Knockout
Mx1-Cre
Induction of Cre in 6-8 week old mice in mainly endothelial
cells. Hemorrhages throughout the brain (cerebrum and
cerebellum at 7-8 months. Lesions dilated vessels with
impaired and disrupted endothelial lumen structure with
microglia recruitment.
(Cunningham
et al., 2011)
Ccm2+/-;
Msh2-/(CCM2,
MSH2)
Ccm2L
(CCM2L)
Mouse
Sensitized
Heterozygous
No overt phenotype. No detectable CCM-like lesions.
(McDonald et
al., 2011)
Mouse
Global
Knockout
No overt phenotype
(Zheng et al.,
2012)
Heg1
(HEG1)
Mouse
Global
Knockout
Embryonic and postnatal lethal due to defective cardiac
integrity and pulmonary hemorrhage. Defective lymphatic
vessel integrity.
(Kleaveland
et al., 2009)
Heg1-/-;
Ccm2+/(HEG1;
CCM2)
Ccm3
(PDCD10)
Mouse
Heterozygous
Died before E10.0. Pericardial edema and aberrant
formation of a dilated aortic sac. No blood circulation due
to a failure of the endothelial cells to lumenize.
(Kleaveland
et al., 2009)
Mouse
Global
Knockout
Embryonic lethal by E8.5, no visible blood cell-filled vessels
in yolk sac, thinner myocardia and smaller ventricular
chambers.
(He et al.,
2010)
Ccm3
(PDCD10)
Mouse
Global
Knockout
Embryonic lethal E 8.0 (prior to the requirement for
cardiovascular function)
(Chan et al.,
2011)
Ccm3
(PDCD10)
Mouse
Glial cell
Knockout;
Emx1-Cre
Survived to adulthood, had enlarged brains due to a
progressive increase in reactive astrocytes. Formation of
CCM-like lesions after 7 months.
(Louvi et al.,
2011)
Ccm3
(PDCD10)
Mouse
Glial cell
Knockout;
Mortality rate of ~80% at age 4-weeks. Enlarged brains
with dilated ventricles and decreased vessel density.
Formation of superficial CCM-like lesions after 3-weeks.
(Louvi et al.,
2011)
Gfap-Cre
Ccm3
(PDCD10)
Mouse
Neural Cell
Knockout;
Nestin-Cre
Viable with normal retinal vasculature.
(He et al.,
2010)
Ccm3
(PDCD10)
Mouse
Neural Cell
Knockout;
Nestin-Cre
No overt phenotype.
(Chan et al.,
2011)
Ccm3
(PDCD10)
Mouse
Neural Cell
Knockout;
Nestin-Cre
Did not survive past postnatal day 3, enlarged brain with
abnormal cytoarchitecture.
(Louvi et al.,
2011)
Ccm3
(PDCD10)
Mouse
Endothelial
Knockout;
Tie2-Cre
Embryonic lethal by E10.5. Fails to remodel vasculature at
E8.5 and has decreased vascular density. Defects in
VEGFR2 signaling and thus defective hematopoiesis.
Dilated and disrupted dorsal aorta and cardinal vein, with
reduced & disrupted cell-cell junctions.
(He et al.,
2010)
Ccm3
(PDCD10)
Mouse
Endothelial
Knockout;
Tie2-Cre
Indistinguishable from littermates at E9.5. Progressive
enlargement of the cardinal vein, rupture/death by E13.5.
(Chan et al.,
2011)
Ccm3
(PDCD10)
Mouse
Smooth
Muscle
Knockout;
Viable with normal retinal vasculature.
(He et al.,
2010)
SM22α-Cre
Ccm3
(PDCD10)
Mouse
Inducible
Knockout;
PDGFb-iCreER
Induction at P1 expresses Cre throughout the brain
vasculature. CCM-like lesions at 1-month of age (fully
penetrant).
(Chan et al.,
2011)
The role of CCM proteins and their binding partners in development. The loss of CCM
genes (KRIT1, CCM2 and PDCD10) is associated with defects in vascular development
and angiogenesis, as well as the formation of vascular lesions. As each of the CCM genes
is required for development, rescue models in zebrafish and conditional/inducible knockout
studies in mice have been paramount for gathering information. This table describes the
phenotypes associated with the loss of CCM genes in zebrafish, fruit-flies and mice.
Animal model information for CCM protein binding partners was only included if the
phenotype overlapped with or helped to inform the CCM-model. The human homolog is
parenthetically noted after each gene reference.