Download Chronic Effects of Inflammatory Cytokines on Cardiovascular and

Considering a career in
translational research?
Babbette LaMarca, Ph.D.
Associate Professor,
Thesis Director, Maternal Fetal Medicine
Department of Pharmacology & Obstetrics & Gynecology
University of Mississippi Medical Center,
Jackson MS
What is translational research?
• Research that helps to make findings from
basic science useful for practical applications
that enhance human health and well-being.
• "translate" findings in basic research quickly
into medical practice and meaningful health
outcomes
“From bench to bedside”
• From laboratory experiments to clinical trials
to point-of-care patient applications.
• All medicinal therapeutics came basic science
that led to a paradigm shift
Tool 1- Ask the right question
• Be able to discern the difference between basic
research performed “for the sake of
knowledge” from what could become
clinically relevant.
Tool 2- Build a multi-skilled team
• Research nurse, to identify patients and enroll
them into a study
• The physician to treat the patient and help with
collection of samples
• The researcher to process samples and either
perform cell culture, run ELISA’s and process
data and run statistics
• Collaborate as a team to write the papers and
communicate your science to the public
Ongoing translational research
at UMMC
Could 17 OHP be a potential therapy for
early gestational age diagnosed
preeclamptics?
Pregnancy-Induced Hypertension
Characteristics
Preeclamptic Women
Hypertension
+
Proteinuria
+
Renal vasoconstriction
+
TPR/ CO
+
Endothelial dysfunction
+
Intrauterine growth restriction
+
TPR, total peripheral resistance.
Preeclampsia is a major cause for
prematurity
• World wide-preeclampsia is responsible for up
to 20% of the 13 million preterm births each
year.
• US-100,000 of the total 500,000 premature
births annually
Pathophysiology of preeclampsia
Fig. 1.
From SM Parikh & SA Karumanchi. Nature Med 2008 14, 810 – 812.
Available at
www.nature.com.proxygw.wrlc.org/nm/journal/v14/n8/full/nm0808810.html
Placental cytokines are increased
in preeclampsia
120
Placental IL-6
pg/ml
100
Placental sFlt-1
pg/ml
500
400
80
300
60
200
40
100
20
0
0
NP
PE
Normoxia (6%)
NP
PE
NP
Hypoxia (1%)
5
PE
Normoxia (6%)
PE
Hypoxia (1%)
Placental IL-10
pg/ml
4
3
LaMarca,
unpublished
data
2
1
0
NP
NP
PE
Normoxia (6%)
NP
PE
Hypoxia (1%)
Leukocyte cytokine secretion is
increased in preeclampsia
LaMarca,
unpublished
data
Preeclampsia is Associated with Chronic
Immune Activation
Chronic innate
Inflammatory Response
Normal
Preeclamptic
+
+++
Monocytes, macrophages,
T helper cells, and possibly NK cells
T helper 17 cells
+
+++
FoxP3CD4+Treg Cells
+
-----
Endothelial dysfunction
-
+++
-
+++
TNF alpha, IL-6, GM-CSF
Activated immune cells
ET-1, adhesion molecules, IL-8, MCP-1
AT1 and ETA autoantibodies
Treatment options for
Preeclampsia
1. I.V. infusion of magnesium sulfate is the therapeutic used for
prevention of seizures during eclamptic events, (>32wks)
Parameters measured by Doppler flow velocity in the uterine,
umbilical and middle cerebral arteries significantly improved in
severely preeclamptic women with Mag.
Administration of MgSO4 significantly decreased systolic, diastolic
and maternal blood pressure as well as decreased resistance index
in uterine arteries
2. Labetalol and hydralazine are the basic anti hypertensives, calcium
channel blockers are strong, alphamethyldopa and furosemide are
used frequently, beta blockers are less frequent
3. ARBs, Etas and ACEi are never used. Several studies show that
they cause fetopathy in RAS and renal development of baby.
Progesterone attenuates TNF-α-stimulated
ET-1 production
Fig. 1.
From Keiser et al. 2009 Am J Hypertension 22: 1120-1125.
Available at
ajh.oxfordjournals.org.proxygw.wrlc.org/content/22/10/1120.full
Circulating progesterone is decreased in
preeclamptic women
Fig. 1.
From Kiprono et al. Am J Obstet Gynecol 2013-04-30
Available at https://www-clinicalkeycom.proxygw.wrlc.org/#!/ContentPlayerCtrl/doPlayContent/1-s2.0S0002937813003451/
Potential use of 17 OHP for
Preeclampsia?
• 17-Hydroxyprogesterone is a natural progestin, produced
mainly by the adrenal glands, gonads, and corpus luteum of
the ovary
• In pregnancy increases in third trimester due to the fetal
adrenal production
• 17-Hydroxyprogesterone Caproate is a synthetic hormone,
similar in structure, interacts with the progesterone receptor
• 17 OHP C is used to prevent subsequent preterm labor
• Considered to be useful in patients diagnosed between 22-30
wks gestation
Administration of progesterone on endothelial cell
ET-1 secretion in response to PE serum
2
6 hours
ET-1
(pg/ml)
1.5
Data from
Kiprono et al. Am
J Obstet Gynecol
2013-04-30
1
0.5
0
35
30
24 hours
25
ET-1
20
(pg/ml)
15
LaMarca,
unpublished
data
10
5
0
NP controls + P
PE patients + P
Progesterone blunts hypoxia
stimulated cytokines and sFlt-1
Placental sFlt-1 expression
24hr Placental Explant Culture
300
5000
Normoxic
Hypoxic
200
N = 4 patients
4000
Normoxic
Hypoxic
sFLT-1 (pg/mg/mL)
IL-6
(pg/mg/mL)
n=4
100
3000
2000
1000
0
0
0.1 uM
1 uM
0
10 uM
0 uM
Concentrations of Progesterone
1 uM
10 uM
Concentration of Progesterone
24hr Placental Explant Culture
24hr Placental Explant Culture
30
5
n=2-4
Normoxic
Hypoxic
3
2
IL-17
(pg/mg/mL)
n=5-6
p=0.051
4
TNF-alpha
(pg/mg/mL)
0.1 uM
20
Normoxic
Hypoxic
10
1
0
0
0
0.1 uM
1 uM
Concentrations of Progesterone
10 uM
0
0.1 uM
1 uM
Concentrations of Progesterone
10 uM
LaMarca,
unpublished
data
Administration of 17-OHP on MAP in response to
placental ischemia or elevated TNF alpha or IL-6
Fig. 1. From Veillion, Am J Obstet
Gyncol 2009, 201:324
Freely available at
www.ncbi.nlm.nih.gov/pmc/article
s/PMC2810642/
and
Fig. 2. From Kiprono et al. Am J
Obstet Gynecol 2013-04-30
Available at https://wwwclinicalkeycom.proxygw.wrlc.org/#!/Content
PlayerCtrl/doPlayContent/1-s2.0S0002937813003451/
Fig. 2, top. From Keiser et al. 2009 Am
J Hypertens 22: 1120-1125.
Available at
ajh.oxfordjournals.org.proxygw.wrlc.or
g/content/22/10/1120.full
*
p=0.059
LaMarca, unpublished data
17 OHP decreases pathophysiology associated with
hypertension during pregnancy
Administration of 17 OHP decreased circulating
levels of TNF alpha and IL-6 in RUPP rats
Fig. 3. From Veillon Jr. et al. Am J
Obstet Gynecol 2009, 201(3):325.e1325.e6.
Available at
www.ncbi.nlm.nih.gov/pmc/articles/P
MC2810642
Progesterone decreases TNF alpha stimulated renal ET-1
Fig. 3A. From Keiser. et al. Am J
Hypertens 2009, Oct 22(10)11201125.
Available at
www.ncbi.nlm.nih.gov/pubmed/1974
5821
Administration of 17 OHP decreased circulating
levels of AT1-AA in IL-6 infused rats
20
15
LaMarca, unpublished data
AT1-AA
bpm 10
5
0
NP
IL-6
NP+17OHP
IL-6+17OHP
Half of Science is Serendipity
• Progesterone attenuates TNF alpha induced ET-1 secretion.
• Administration of progesterone decreased endothelial cell
activation in response to serum from PE women
• Administration of progesterone (1uM) significantly blunts
hypoxia stimulated inflammatory cytokines and sFlt-1 from
placental explants
• Administration of 17 OHP decreased blood pressure in
response to RUPP or elevated TNF alpha or IL-6 during
pregnancy
• Administration of 17 OHP decreased circulating cytokines in
RUPP rats, local ET-1, and circulating AT1-AA in response to
IL-6 infusion
Skill sets to acquire
• 1. Learn to interpret the data and ask clinically
relevant questions
• 2. How can that data be applied to a
pathophysiological condition
• 3. Communicate your data and ideas with
physicians and discuss a potential
collaboration and clearly define the clinicians
role in the research
Skill sets to acquire
• 4. Build your research team
• 5. Submit your IRB and start collecting
samples
• 6. Communicate with your team regularly with
data updates and writing the papers and grants.
Your communication with the
practicing physician and nurses is
key to successful translational
research
Translational research is the key
to improving quality of patient
care