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D9 Poster Session VIII - Group D: Vascular
D9-01
DEVELOPMENT OF ROCK2-SELECTIVE BA-1049 FOR
TREATMENT OF CEREBRAL CAVERNOUS MALFORMA-
TIONS
Lisa Bond
, Kenneth Rosen, Lisa McKerracher
BioAxone BioSciences, Inc., Research, Cambridge, USA
Cerebral cavernous malformation (CCM) disease is a cerebrovascular
disorder in which endothelial cells form single or multiple cystic brain
lesions that can leak and cause hemorrhagic stroke and other neurological
defects. There is no pharmacological treatment to prevent/reverse CCM
lesion formation; severe clinical sequelae are treated by surgical removal
of the leaky capillary clusters, potentially causing additional neuro-
trauma. Inherited cases of CCM are caused by loss of function in one of
the 3 CCM genes (
CCM1, CCM2 and CCM3
) and sporadic cases result
from mutations in the same genes. Studies of the inherited mutations
underlying CCM formation revealed that disruption in endothelial barrier
integrity is caused directly by the hyper-activation of Rho, a small
GTPase signaling protein that regulates stress fiber formation and cell-
cell junctions. We previously demonstrated abnormal activation of Rho
after SCI and TBI, and developed BA-1049 as a compound that reverses
abnormal activation of downstream Rho kinase (ROCK) in neurotrauma.
BA-1049 is selective for the Rock2 isoform highly expressed in cerebral
vasculature. To investigate potential efficacy of BA-1049 in CCM, we
studied the effect of BA-1049 on endothelial cells. BA-1049 reduces
proliferation in human vascular endothelial cells (HUVEC)
in vitro
,
suggesting a potential to reverse the angiogenic abnormalities underlying
CCM formation. Treatment with BA-1049 also disrupts stress fiber for-
mation and increases occludin/cadherin expression in HUVEC cells,
suggesting a potential to reverse the junctional instability and endothelial
permeability underlying lesion hemorrhage. To determine if BA-1049
has appropriate properties to be used as an orally active drug, we in-
vestigated Caco-2 cell permeability and albumin binding. BA-1049 has
good permeability characteristics (Papp Ratio: 1.31) and shows low al-
bumin binding (7.26%). We have performed a computer-based screen of
the chemical structure, and no hits indicating potentially serious toxicities
were detected. We have focused on CCM for development of BA-1049
because of the clear clinical development path. Success in development
of BA-1049 for CCM may lead to further development to reverse Rho
activation in other types of neurotrauma, such as TBI.
Keywords: Cerebral Cavernous Malformations, Rho Kinase, Vas-
cular Disorders, CCM, ROCK
D9-02
HUMAN UMBILICAL CORD PERIVASCULAR CELL
(HUCPVC) THERAPY FOR TRAUMATIC BRAIN INJURY:
TARGETING THE NEUROVASCULAR UNIT
Tanya Barretto
2
,
Eugene Park
1
, Leila Maghen
2
, Elaine Liu
1
, Shlomit
Kenigsberg
2
, Andree Gauthier-Fisher
2
, Katya Park
2
, Andrew Baker
1,3
,
Clifford Librach
2,4
1
St. Michael Hospital, Trauma Research, Toronto, Canada
2
CReATe Fertility, Research, Toronto, Canada
3
University of Toronto, Surgery & Anesthesia, Toronto, Canada
4
University of Toronto, Obstetrics & Gynecology, Toronto, Canada
Mesenchymal stem cells are currently being used in clinical trials for
treatment of neurodegenerative diseases including stroke. We were
interested in evaluating the potential of human umbilical cord-derived
perivascular cells (HUCPVCs), a less differentiated type of MSC, on
their therapeutic potential in TBI. HUCPVCs have several advantages
over MSCs including ease of procurement and greater rates of ex-
pansion.
Objective:
We hypothesized that HUCPVCs contribute to recovery
of white matter after modeled trauma.
Methods:
A gene array analysis was performed on HUCPVCs to
examine expression of relevant modulators of inflammation, angio-
genesis and neurogenesis.
In vitro
and
in vivo
injury models were used
to evaluate HUCPVC treatment on outcome after injury.
In vitro
-
E17-derived cortical neurons were cultured for 7 days, then subject to
sublethal oxygen-glucose deprivation for 90 minutes resulting in ax-
onal degeneration. HUCPVCs were co-cultured for 3 days and out-
come assessed on degenerating axons.
In vivo
- Adult male Sprague-
Dawley rats underwent a lateral fluid percussion injury and were
treated 24 hours after injury with 2.5x10
6
fluorescently labeled HUC-
PVCs via tail vein injection.
Results:
Gene expression analysis indicates expression of key
neurotrophic (BDNF, NGF, NT3, GDNF), angiogenic (VEGF-A,
FGF5, BMP1) and inflammation-modulating factors (CSF, IL mem-
bers, CXCL members). Preliminary
in vitro
data indicates that
HUCPVCs rescue OGD-induced axonal degeneration.
In vivo
results
indicate no adverse effects related to HUCPVC treatment. Labeled
HUCPVCs were found in the injured cerebral cortex at 24 hrs post-
injection.
Conclusions:
Our data suggests that HUCPVCs have therapeutic
potential to address white matter injury after TBI. Given the immune
modulating expression of cytokines, proangiogenic and neurotrophic
factor expression there is evidence to suggest that HUCPVCs targets
numerous injury mechanisms.
Keywords: stem cell therapy, human umbilical cord perivascular
cell, mesenchymal stem cell
D9-03
INSTITUTIONAL REVIEW OF SCREENING FOR BLUNT
CEREBROVASCULAR INJURIES
Matt Decker
, Greg Murad
University of Florida, Neurosurgery, Gainesville, USA
Introduction:
The incidence of blunt cerebrovascular injuries (BCVI)
in the trauma population is low; however, the mortality and morbidity
associated with symptomatic BCVI approaches 30% and 60% re-
spectively. Appropriate screening methods need to be implemented to
ensure treatment is initiated prior to symptom development. Screening
algorithms rely mostly on level III evidence. This study evaluates a
single institution’s data at a Level I trauma center to develop insti-
tutional neurosurgical guidelines screening for BCVI.
Methods:
IRB for a retrospective review was obtained. The Uni-
versity of Florida Trauma Database was queried. All patients pre-
senting to UF Health since its designation of a Level I Trauma Center
( July 1, 2005) with ICD-9 diagnosis codes specific to neurosurgical
trauma were individually chart reviewed. Patients included were those
who underwent neurovascular imaging and those with a diagnosis of a
blunt cerebrovascular injury. Exclusion criteria included patients with
a documented history of prior cerebrovascular injury, those with
penetrating cerebrovascular injuries, and patients undergoing vascular
imaging to assess for intracranial vascular malformation (aneurysm,
AVM). Data collected included patient demographics; traumatic in-
juries; signs, symptoms, treatment, and complications of BCVI; and
follow up. A multivariate analysis was performed to determine odds
ratios based on trauma findings.
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