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International Journal of Anesthesiology & Pain Medicine
ISSN: 2471-982X
Volume 4
March 26-28, 2018
Vienna, Austria
Pain Management 2018
Internal Medicine 2018
Page 39
JOINT EVENT
7
t h
E d i t i o n o f I n t e r n a t i o n a l C o n f e r e n c e o n
Internal Medicine and Patient Care
&
6
t h
E d i t i o n o f I n t e r n a t i o n a l C o n f e r e n c e o n
Pain Management
Background:
Sepsis microvascular dysfunction embraces
different cellular components including endothelial cells, in
which it increases its permeability and activation to shed extra
microparticles (MPs) to transport a unique cellular signaling to
the recipient cells. In this study, we observed that microparticles
can retain different epigenetic components as miRNA, mRNA
of DNMTs and HDACs from parent cells that can transfer to
naïve target cells. Importantly, in sepsis, MPs production is
increased. Increased expression of DNMTs results in promoter
hypermethylation which can suppress transcription of not only a
single gene but networks of genes with systemic effects. Sepsis
is an inflammatory insult which can result in vascular dysfunction
leading to systemic shock and eventual death.
Aim:
The aim of this study is to distinguish the role of sepsis
microparticles in systemic immunosuppression process and
the impact of these particles upon cellular targets and survival
mechanisms to allow better diagnostic tools and potential novel
therapeutic approach during infection and trauma.
Methodology:
Endothelial cells ( HUVEC) and naïve monocytes
treated with MPs from patients with sepsis demonstrated
dramatically reduced of anti-inflammatory genes, TGF-β, TNF-α
expression and some of autophagy molecules (ATG5, ATG7
and LC3) due to hypermethylation of their promoter. These data
demonstrate that mRNAs of epigenetic regulators including
DNMTs are highly expressed in plasma MVs in patients with
sepsis and can be transferred to naïve cells through MVs and
cause pro-inflammatory cytokine gene silencing and autophagy
repression in monocytes. Further, MVs per mL plasma on day 1
alone significantly correlated with death by day 5 (r=0.7125 and
p=0.0042). Using immunostaining techniques and flow cytometer,
we found the major source of plasma MVs in the critically-ill, non-
septic control patients shifted frommonocytes (Mo) to endothelial
cells (EC) in the SS patients (Control: Mo 63.6% and EC 7.4% and
SS: Mo 12% and EC 58.7% qualitatively). Focusing our study on
SS patients who lived and SS patients who died by day 5, our data
shows that while total DNMTmRNA copy numbers per plasmaMV
are significantly higher over days 1 and day 3 in those SS patients
who lived, the ratios of DNMT1 (maintenance DNA methylation)
and the combination DNMT3A and DNMT3B (de novo DNA
methylation) are reversed on days 1 and 3 (SS Lived: DNMT3A/3B-
to-DNMT1: day 1=0.68 and day 3=0.87; SS Died: DNMT3A/3B-to-
DNMT1: day 1=2.49 and day 3=2.94). Finally, MV DNMT3A/3B
mRNA from day 1 samples positively correlates with reduced
survival (r=6261 and p=0.0165). Targeting of circulating MVs with
commercially available inhibitors of DNMTs may be a therapeutic
strategy in specific patients with deregulated epigenetic
mechanisms to limit both early and chronic consequences.
Results:
We found that MPs from patients with septic shock and
septic had significantly increased mRNA for DNMTs compared
to MPs from patients with critical illness without sepsis and from
normal healthy adults over the course of 5 days. Remarkably,
we noticed that DNMT1 and -3a mRNA has the highest gene
expression in sepsis MPs compared to other DNMTs. Additionally,
naïve monocytes treated with MPs from patients with sepsis
demonstrated increased expression of DNMTs. At the same time
decreased expression at 24 hours.
Epigenetic re-programming of plasma
microvesicles in sepsis
Duaa Dakhlallah, Jon Wisler, Tierra Ware, Erin
Leatherman, Yijie Wang, Amy Gross, Joyce Obeng,
Ahmad Dakhlallah, Timothy D Eubank and Clay B
Marsh
West Virginia University, USA
Duaa Dakhlallah, Int J Anesth Pain Med 2018, Volume 4
DOI: 10.21767/2471-982X-C1-001