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com
BCS Abstracts 2014
LOS groups had significant reductions in diastolic blood pressure.
In addition, the PRO group showed reduced basal HR (373 bpm
± 13  421 bpm ± 42), and the ENL group presented reduced
intrinsic pacemaker HR (308 bpm ± 26  343 bpm ± 30). The
vehicle group showed sympathetic dominance in determining the
basal HR, in contrast, the PRO group showed vagal dominance in
determining the baseline HR when compared to the ENL, HCTZ
and AML groups, considering that for these animals the chrono-
tropic response after the administration of propranolol was signifi-
cantly lower, when compared to the response obtained after the
administration of atropine. For HRV, the pharmacologically
treated group showed no significant difference in the values of
low frequency band (LF; 0.2–0.75 Hz) and higher power in high
frequency band (HF; 0.75–3.0 Hz), compared to the vehicle
group. The analysis of systolic arterial pressure variability revealed
that the ENL group showed a reduction in the LF band, compared
to the vehicle, LOS, PRO and HCTZ groups.
Conclusion None of the pharmacological treatment was able to
completely attenuate the adverse effects of hypertension on the auto-
nomic parameters in spontaneously hypertensive rats, however, the
group treated with Enalapril showed a positive effect on the SAPV.
177 THE ALPHA-KETOGLUTARATE RECEPTOR GPR99
REGULATES PATHOLOGICAL CARDIAC HYPERTROPHY
Ameh Omede*, Delvac Oceandy, Mamas Mamas, Elizabeth Cartwright, Min Zi,
Sukhpal Prehar, Arfa Maqsood. Institute of Cardiovascular Sciences, University of
Manchester
10.1136/heartjnl-2014-306118.177
Introduction GPR99, a member of G-protein coupled receptor
family, is expressed in the heart. Previous studies suggested that
this receptor can bind to alpha-ketoglutarate, a metabolite that is
elevated in the serum of heart failure (HF) patients. However,
the functional role of GPR99 in cardiomyocytes is unknown. In
this study, we investigated whether GPR99 regulates cardiac
hypertrophy, a key process in the development of HF.
Results Mice with genetic ablation of GPR99 (GPR99-/-) dis-
played an increased in hypertrophy following transverse aortic
constriction (TAC) as indicated by heart weight/body weight
ratio. Furthermore, GPR99-/- mice showed significantly increased
Abstract 178 Figure 1 Box plots showing changes miR-483-3p expression and NT-proBNP levels after 6 months LVAD therapy. There is
upregulation of plasma (A) and myocardial (B) miR-483-3p expression, with fold change 2.32 (1.30-2.44;p = 0.021) and 1.799 (0.717-4.719; p =
0.169) respectively. NT-proBNP shows a corresponding significant reduction. Outlier results are shown with -.
*Significant to p
A100
interstitial fibrosis and larger cell surface area compared to wild-
type controls. The cardiac function as indicated by fractional
shorting (FS) is significantly lower in the knockout mice compared
to wildtype mice following TAC. To examine the mechanism we
performed yeast two hybrid screening analysis and have identified
CSN5, a member of the COP9 signalosome complex, as a novel
interacting partner of the GPR99 receptor. COP9 is known as a
regulator of protein degradation via the ubiquitin proteasome sys-
tem. Adenoviral mediated overexpression of GPR99 in cardio-
myocytes induced the ubiquitination and degradation of a
prohypertrophic factor interferon regulatory factor 5 (IRF5). Con-
sistently, in GPR99-/- mice expression of IRF5 was significantly
increased following TAC, which might provide the possible mech-
anism responsible for the increased hypertrophy in these mice.
Conclusion Our findings suggest that the alpha-ketoglutarate
receptor GPR99 modulates pathological hypertrophic response
by modulating ubiquitination of IRF5. Thus, GPR99 may
become the possible target for heart failure treatment.
178 CIRCULATING MICRORNAS FOR PREDICTING AND
MONITORING RESPONSE TO MECHANICAL
CIRCULATORY SUPPORT FROM A LEFT VENTRICULAR
ASSIST DEVICE
1
Andrew Morley-Smith*, 2Adam Mills, 3Steven Jacobs, 3Bart Meyns, 3Filip Rega,
1
Andre Simon, 1John Pepper, 1Alexander Lyon, 4Thomas Thum. 1Imperial College London
and Royal Brompton/Harefield; 2Imperial College London; 3University Hospitals Leuven;
4
Hannover Medical School and Imperial College London
10.1136/heartjnl-2014-306118.178
Purpose There are few non-invasive techniques to predict and
monitor patients’ responses to left ventricular assist device
(LVAD) therapy. MicroRNAs (miRs) are small noncoding RNAs
with intricate roles in cardiovascular disease. They remain stable
in the circulation, are readily quantified, and may be useful as
new biomarkers. This study sought to identify candidate miR
biomarkers for further investigation and to investigate whether
these circulating miRs were of myocardial origin.
Methods and Results We studied 55 serial plasma and myocar-
dial samples from 19 patients who underwent HeartMate II
LVAD implantation, and used a screening microarray to analyse
Heart 2014;100(Suppl 3):A1–A138
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the change in expression of 1,113 miRs from pre-implant and 6
months follow-up. Twelve miRs showed significant variation and
underwent validation, with miR-1202 and miR-483–3p selected
for further study. The key findings in the test cohort (n = 8)
were: (1) miR-483–3p showed upregulation after 6 months of
LVAD therapy in circulating plasma (median fold change 2.32
(1.30–2.44); p = 0.021) and in ventricular myocardium (median
fold change 1.799 (0.717–4.719; p = 0.169). This mirrored the
reduction in NT-proBNP levels at 6 months (fold change 0.30
(0.08–0.43); p = 0.004; see Figure). (2) Using change in NT-
proBNP at 3 months as a marker of clinical response to LVAD
therapy, baseline expression of plasma miR-1202 identified good
versus poor LVAD responders (absolute expression 1.296
(1.293–1.306) vs. 1.311 (1.310–1.318) A.U.; p = 0.004). (3)
Both miR-483–3p and miR-1202 are also enriched in ventricular
myocardium suggesting the heart as the possible source of these
plasma miRs.
Conclusions This is the first report of circulating miR bio-
markers in LVAD patients. We demonstrate the feasibility of this
approach, and report the potential for miR-483–3p and miR-
1202 respectively to monitor and predict response to LVAD
therapy. We suggest that changes in miR-483–3p expression
could provide a more specific assessment of ventricular function
that complements the changes in systemic neuroendocrine milieu
recorded by serial measurement of natriuretic peptides, and sug-
gest that miR-1202 could be valuable for judging the likelihood
of good response to LVAD support. However, these conclusions
are weakened by small sample size and retrospective analysis. We
propose further work to study these hypotheses further and elu-
cidate roles for miR-483–3p and miR-1202 in clinical practice
and in underlying biological processes.
179 THE ROLE OF MICROPARTICLES IN CAROTID DISEASE
1
Andrew Schiro*, 2Fiona Wilkinson, 2Ria Weston, 3JVincent Smyth, 1Andrew JBoulton,
3
Ferdinand Serracino-Inglott, 2Yvonne MAlexander. 1
University of Manchester;
2
Manchester Metropolitan University; 3Manchester Royal Infirmary
10.1136/heartjnl-2014-306118.179
Introduction Endothelial microparticles (EMPs) are released
from dysfunctional endothelial cells. We hypothesised that
patients with unstable carotid plaque have higher levels of circu-
lating microparticles compared to patients with stable plaques
which could be related to a specific cytokine profile.
Methods Circulating EMPs and inflammatory cytokine levels
were measured in seventy patients with significant carotid disease
undergoing carotid endarterectomy and 20 healthy controls. Fifty
one (73%) patients had symptomatic disease whilst 19 (27%) were
asymptomatic. EMPs (CD31+/ Annexin V+ CD42b-) were quanti-
fied using flow cytometry. Immunohistological analysis of carotid
plaques for CD68+, CD206+ macrophages, TNF-a smooth
muscle actin and osteopontin was performed, together with Ali-
zarin red staining for detection of calcific deposits. Bioplex assays
were used for cytokine analysis. Plaques were graded according to
the American Heart Association plaque scoring system.
Results Significantly higher EMP levels were observed in sympto-
matic patients compared to controls, p = 0.01, while no differen-
ces were noted in EMP levels in asymptomatic vs controls p =
0.11. The higher EMP levels appeared to associate with the unsta-
ble plaques which also had a significantly higher level of CD68+
macrophages compared to stable plaques (AHA I-IV) and higher
circulating levels of Chemokine ligand-9 (CXCL-9) (p < 0.004).
Of note, macrophage inhibitory factor (MIF) was among the
Heart 2014;100(Suppl 3):A1–A138
BCS Abstracts 2014
chemokines that were elevated in the circulation of patients with
stable plaques, whether a lack of this factor in unstable plaques has
direct effects on phenotype remains to be elucidated. Other factors
elevated in patients with stable plaques were IL-16, cutaneous T-
cell attracting chemokine (CTACK), and stem-cell growth factor-b
(SCGF-b) (p value of 0.05, 0.035 and 0.002 respectively).
Conclusion Circulatory EMP and specific inflammatory cytokine
levels are raised in patients with unstable plaques, while MIF, a
potentially protective factor was elevated in serum of patients with
stable plaques. These data could have major implications for the
development of a diagnostic tool whereby EMPs together with
markers of macrophage activity could act in a combined manner
as biomarkers of plaque vulnerability and stroke susceptibility.
180 CAN NEUTROPHIL APOPTOSIS BE TARGETED
THERAPEUTICALLY TO MODIFY INFLAMMATION
DURING MYOCARDIAL INFARCT HEALING?
Xiaofeng Zhao*, David Dorward, Adriano Rossi, Gillian Gray. University of Edinburgh
10.1136/heartjnl-2014-306118.180
Neutrophils are rapidly recruited to infarct site in response to
cardiomyocyte death. Apoptosis and phagocytosis by macro-
phages prevents further damage to host tissues, but can also
polarise macrophages towards an anti-inflammatory phenotype,
potentially enhancing repair and reducing infarct expansion after
myocardial infarction. Pharmacological induction of neutrophil
apoptosis by cyclin-dependent kinase inhibitor drugs (CDKi) has
been shown to enhance resolution of inflammation in the murine
lung (Rossi et al ., 2006). The present study study investigated
the in vitro effects of a CDKi (AT7519) on mouse neutrophil
apoptosis and determined its effects on agonist-induced Ca2+
flux.
Immature bone marrow-derived neutrophils (BMDNs) were
isolated and inflammatory neutrophils obtained from the lavage
of thioglycollate-induced peritonitis (TGNs). Apoptosis was
assessed by flow-cytometric analysis of annexinV/propidium
iodide (PI) binding, DNA incorporation of PI into permeablised
cells (hyplodiploid peak) and morphological assessment of cyto-
centrifuge preparations. Intracellular Ca2+ flux was assessed by
spectrofluorimetric analysis of Fura-2 loaded cells.
BMDNs and TGNs underwent constitutive apoptosis at 6 and
20h, an effect enhanced by AT7519 in a concentration, time and
caspase-dependent manner. For example, at 6h, apoptosis
assessed by hypodiploid peak quantification was 6.7% in control
BMDNs vs 15.2% in 1 µM AT7519 treated cells. In TGNs,
apoptosis as assessed by annexinV/PI binding at 6 h were 6.2%
in control vs 12.0% in 1µM AT7519 treated cells (n = 6). Inter-
estingly, AT7519 did not directly induce Ca2+ fluxes in BMDNs
or TGNs nor did it affect Ca2+ fluxes triggered by neutrophil
agonists (fMLF and PAF).
Thus AT7519 enhances mouse neutrophil apoptosis without
affecting early activation responses such as agonist-induced Ca2+
flux and therefore suggests that CDKi induce apoptosis even in
the presence of stimuli found at inflammatory sites, which make
it a promising anti-inflammatory agent to be used in the setting
of MI.
This study was supported by a Chinese Scholarship Council/
University of Edinburgh Fellowship to XZ and by a BHF Centre
of Research Excellence Award.
REFERENCE
Rossi AG, et al. Cyclin-dependent kinase inhibitors enhance the resolution of inflam-
mation by promoting inflammatory cell apoptosis. Nature Med 2006;12:1056–1064
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178 Circulating Micrornas for Predicting and

Monitoring Response to Mechanical Circulatory
Support from a left Ventricular Assist Device
Andrew Morley-Smith, Adam Mills, Steven Jacobs, Bart Meyns, Filip Rega, Andre
Simon, John Pepper, Alexander Lyon and Thomas Thum

Heart 2014 100: A100-A101

doi: 10.1136/heartjnl-2014-306118.178

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