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GFAP levels were elevated in post-injury samples from all 13 in-
jured micropigs. GFAP levels appear to be elevated as early as 1–
30 min after injury, but 1–6 hr post-injury samples have the highest
GFAP elevations overall. GFAP levels were elevated to a much
lesser extent in the sham-operated micropigs. Serum UCH-L1 pro-
files in micropigs appears more complex. Four of 16 micropigs have
high baseline pre-surgery serum UCH-L1 levels (
>
200 pg/mL) and 5
micropigs have no detectable UCH-L1 signal either pre-surgery or
after injury. Serum Iba-1 levels are modestly elevated post-injury in
only 4 of 12 animals at one or more time points and elevated post-
sham operation in one of 3 animals. Lastly, IL-6 levels were sig-
nificantly elevated in 9 of 11 animals with cFPI at one or more post-
injury time points.
Group analysis showed post-injury GFAP levels significantly el-
evated at 1, 3 and 6h post–injury when compared to pre-injury
controls. There is also a trend of IL-6 increase following FPI. Taken
together, we have identified and characterized several biomarkers
with potential use for future screening of therapeutic agents in this
micropig model.
Supported by DoD grant W81XWH-10-1-0623
Keywords: fluid percussion injury
A5-14
A SYSTEMATIC RAT SPINAL CORD INJURY BIOMARKER
STUDY AND TEMPORAL BIOMARKER PROFILING IN
TWO BIOFLUID TYPES
Zhihui Yang
1
, Helen M. Bramlett
2
, Ahmed Moghieb
1
, Dongnan Yu
1
,
Ping Wang
1
, Fan Lin
1
, Carl J. Bauer
1
, Tyler M. Selig
1
, Zhiqun Zhang
1
,
Ronald L Hayes
3
, Michael Y. Wang
2
, W. Dalton Dietrich
2
, Kevin K.
W. Wang
1
1
University of Florida, Center for Neuroproteomics & Biomarkers
Research, Departments of Psychiatry & Neuroscience, Gainesville,
USA
2
University of Miami, Department of Neurological Surgery, Leonard
M. Miller School of Medicine, Miami, USA
3
Banyan Biomarkers, Inc.,, Banyan Laboratories, Alachua, USA
In the United States, there are approximately 12,000 new cases of
spinal cord injury (SCI) each year and some 1.2 million people living
with paralysis due to SCI. However no effective therapy to treat acute
SCI is available. Here we report a systematic rat SCI biomarker study
examining the effects of injury severity and temporal profiling of
eleven SCI candidate biomarkers in two biofluid types (cerebrospinal
fluid (CSF) and serum). We utilized an established rat thoracic con-
tusive spinal cord injury model with either moderate or severe injury.
CSF and serum samples were obtained at 4, 24 hr, and 7 day post-
injury. Candidate biomarkers we focused on included axonal injury
markers
a
II-spectrin breakdown products (SBDPs), neuronal cell body
injury marker UCH-L1, gliosis/glial injury markers S100
b
& GFAP
and GFAP-BDPs, demyelination marker MBP, neuroinflammation
marker IL-6 and autoantibodies to GFAP and spinal cord protein. Our
results shows three biomarker pattern types: (i) biomarkers that are
severity-dependently elevated in both CSF and serum at acute time
points (4 – 24 h) – SBDPs, GFAP, G-BDPs, S100b and UCH-L1; (ii)
biomarker that is only elevated in serum (4–24 h) but not CSF - IL-6,
suggesting a systemic response to SCI; (iii) biomarkers that appear not
elevated in either biofluid after SCI - MBP and autoantibodies. These
results not only allow us to gain important insight into the patho-
mechanisms of SCI, but our learning from them will directly translate
into human SCI studies.
Keywords: SCI, biomarkers, spectrin, GFAP, S100b, UCH-L1
A5-15
HUMAN SPINAL CORD INJURY CSF AND SERUM BIO-
MARKER STUDY
Michael Y. Wang
2
,
Kevin Wang
1
, Zhihui Yang
1
, Ahmed Moghieb
1
,
Helen Bramlett
2
, Dongnan Yu
1
, Ping Wang
1
, Fan Lin
1
, Carl Bauer
1
,
Tyler Selig
1
, Zhiqun Zhang
1
, Ronald Hayes
3
, M. Ross Bullock
2
, W.
Dalton Dietrich
2
1
University of Florida, Psychiatry/Neuroscience, Gainesville, FL,
USA
2
University of Miami, Neurological Surgery, Miami, FL, USA
3
Banyan Biomarkers, Banyan Laboratories, Alachua, FL, USA
Spinal cord injury (SCI) is a devastating insult and frequently occurs
in young men and women as a result of accidents during sports and
other activities. It also has high frequency in the military among
servicemen. In these cases, SCI can result from blast injuries or high
velocity metal fragments. There are of an estimated 12,000 new cases
of SCI in the United States annually and approximately 1.2 million
people living with paralysis due to SCI. Although much information
has been gained from experimental and clinical studies elucidating
injury mechanisms and the testing of new treatments to target spinal
cord injury, no clinically effective treatments are currently available
to treat SCI. The heterogeneous nature of spinal cord injury may
require customized therapies specific for selective injuries and sub-
jects. We hypothesize that it will be extremely advantageous if bio-
markers sensitive to injury severity and treatment responses can be
identified.
Here we report a systematic human SCI biomarker study examining
the effects of injury severity and temporal profiling of eleven (11) SCI
candidate biomarkers in both cerebrospinal fluid (CSF) and serum.
Serial CSF and serum samples were obtained from 14 SCI patients
with initial ASIA score (AIS) of A-B at every 6 hours for up to 6 days.
SCI biofluid and control CSF (n
=
20) and serum (n
=
20) samples
were subjected to immunoblotting and/or immunoassays for protein
biomarker levels. Candidate biomarkers included axonal injury
markers
a
II-spectrin breakdown products (SBDP150, SBDP145 &
SBDP120), neuronal cell body injury marker UCH-L1, gliosis/glial
injury markers S100
b
, GFAP and GFAP-BDPs, demyelination marker
MBP, neuroinflammation marker IL-6 and autoantibodies to GFAP
and spinal cord proteins. Our key findings are as follows. (i) All brain-
derived protein biomarkers are elevated in the acute phase CSF and
serum samples from SCI patients when compared to corresponding
normal controls. (ii) CSF GFAP-and GFAP-BDP levels (and to a
lesser extent S100b) shows a strong SCI severity correlation. (iii)
Elevation of both CSF UCH-L1 and SBDPs are detected in almost all
SCI subjects. Notably, CSF UCH-L1 shows some unique and late
rises (day 4–5) in a subset of patients. (iv) Of all the serum markers,
GFAP levels has the best correlation with SCI severity (AIS), fol-
lowed by S100b and SBDP150. (v) Serum UCH-L1 tends to peak
early post-injury but also shows sustained levels in some patients on
day 4–6. IL-6 rises acutely in most SCI patients, but its levels appear
independent of SCI severity. Lastly, Anti-GFAP and anti-spinal cord
proteins (IgM, IgG) shows a time-dependent elevation (from day 0 to
6) in serum and to a lesser extent in CSF in subset of SCI patients,
regardless of their injury severity. These human SCI biomarker results
allow us to gain important insight into the pathomechanisms of SCI.
Our results will expand into further human SCI studies by not only
confirming the utilities of biomarkers in monitoring human SCI pro-
gression or recovery but also guiding personalized therapeutic and
rehabilitation strategies.
Supported by grant W81XWH-12-1-0277 from the United States
Army
Keywords: theranostic, GFAP, IL-6, UCH-L1, spinal cord injury
A-34