Download Cellular mechanisms in pulmonary arterial hypertension: focused on prostacyclin and endothelin pathway

Cellular mechanisms in pulmonary arterial
hypertension: focused on prostacyclin and
endothelin pathway
Rijan Gurung standing in for Dr Jigisha Patel
Research associate for Prof. Lucie Clapp
Pulmonary Arterial Hypertension
•
Progressive fatal condition
–
–
Elevated mean pulmonary arterial pressure ≥ 25mm Hg at rest or ≥ 30mm Hg with
exercise.
Reduction in gas exchange, In situ thrombosis, Plexiform lesion formation
 Right ventricular (RV) hypertrophy  Followed by heart failure eventually DEATH
anti/platelet
Humbert M, et al. NEJM. 2004.
Lab Expertise
 Isolation of different cell types
- SMC, ECs, fibroblasts, microparticles etc…
Smooth muscle
microparticles
 Functional assays
- Proliferation, myography, organ-bath etc…
 Molecular biology and pharmacology
 Animal models of PAH
0.75
105
104
10
3
10
2
101
10
3
10
2
101
101
102
103
104
Forward Scatter
10% FBS
 General lab techniques
- WB, ELISA, PCR, qPCR, FACs
8.21
104
FITC - Annexin V
 Biochemistry
105
FITC- Annexin V
 Immunostaining
- IHC, IF
105
101
102
103
104
105
Forward Scatter
10% FBS + 20ng/ml PDGF-BB
Isolation & Characterization of PAECs and PASMCs from
human lungs
A
HPASMCs
α-SMA
SM-22
B
HPAECs
VWF
CD-31
NORMAL
PAH
Combination therapy and drug targets
PASMC PROLIFERATION
Prostanoid receptor expression in PAH vs Normal
120
Normal
0.04
100
PAH
Relative gene expression
80
60
T r e p r o s t in il
+ T a d a la f il
+ R io c ig u a t
0.0030
lo g [ T r e p r o s t in il] M
-5
Prostaniod receptor subtype
P1
-6
D
-7
4
-8
EP
-9
3
-1 0
EP
-1 1
2
0.0000
-1 2
EP
0
-1 3
+ B o s e n ta n
0.0060
1
20
0.02
EP
40
0.03
IP
% C e ll p r o lif e r a t io n
***