a relationship between inflammatory cytokine receptors and
TRANSCRIPT
A relationship between inflammatory cytokine receptors and ion channel expression in endometriotic lesions
Universitätsklinik für Frauenheilkunde [email protected]
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Universitätsklinik für Frauenheilkunde
Endometriotic-associated nerve fibers
A B
C D
E F
McKinnon B, Bersinger NA, Wotzkow C, Mueller MD. Endometriosis-associated nerve fibers, peritoneal fluid cytokine concentrations, and pain in endometriotic lesions from different locations. Fertil Steril. 2012 Feb;97(2):373–80.
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Universitätsklinik für Frauenheilkunde
Cytokine concentrations and endometriosis associated nerve
McKinnon B, Bersinger NA, Wotzkow C, Mueller MD. Endometriosis-associated nerve fibers, peritoneal fluid cytokine concentrations, and pain in endometriotic lesions from different locations. Fertil Steril. 2012 Feb;97(2):373–80.
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Universitätsklinik für Frauenheilkunde
Inflammation and nerve fiber interaction in endometriotic pain;
McKinnon BD, Bertschi D, Bersinger NA, Mueller MD. Inflammation and nerve fiber interaction in endometriotic pain. Trends Endocrinol Metab. 2015 Jan;26(1):1–10.
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Universitätsklinik für Frauenheilkunde
Study aim;
-Determine the expression of cytokine receptors and ion channels in endometriotic tissue, the relationship between them and their cellular localisation;
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Universitätsklinik für Frauenheilkunde
No Endo Endo Peritoneal Ovarian RVS
0.03
1.00
32.00
rela
tive
mR
NA
exp
ress
ion
TNFR1
Eutopic Ectopic
*** ***
Cytokine receptors and ion channel mRNA expression in endometrial and endometriosis tissue;
Cytokine Receptors
Purinergic Receptors Voltage gated sodium channel
No Endo Endo Peritoneal Ovarian RVS
0.03
1.00
32.00
rela
tive
mR
NA
exp
ress
ion
CCR5
*** *** *
Eutopic Ectopic
No Endo Endo Peritoneal Ovarian RVS
0.03
1.00
32.00
rela
tive
mR
NA
exp
ress
ion
SCN9A
** ***
Eutopic EctopicNo Endo Endo Peritoneal Ovarian RVS0.03
1.00
32.00
1024.00
rela
tive
mR
NA
exp
ress
ion
P2X3
Eutopic Ectopic
No Endo Endo Peritoneal Ovarian RVS
0.03
1.00
32.00
rela
tive
mR
NA
exp
ress
ion
P2X4
Eutopic Ectopic
*** ***
Endo Non-Endo
Eutopic Ectopic
P O RVS
E NE
Eutopic Ectopic
P O RVS No Endo Endo Peritoneal Ovarian RVS
0.03
1.00
32.00
rela
tive
mRN
A e
xpre
ssio
nSCN3A
*** ***
Eutopic EctopicE NE
Eutopic Ectopic
P O RVS E NE
Eutopic Ectopic
P O RVS E NE
Eutopic Ectopic
P O RVS
Endo Non-Endo
Eutopic Ectopic
P O RVS
P2X3 P2X4
CCR5 TNFR1
SCN3A SCN9A
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Universitätsklinik für Frauenheilkunde
Relationship between cytokine receptors and ion channels in ectopic lesions;
Correlation; CCR5 v Purinergic Receptors Correlation; CCR5 v Voltage gated sodium channel SCN9A
0 1 2 3 4 50.00
5.00
10.00
15.00Linear reg. of CCR5:Line
1 2 3 4 5-5.00
0.00
5.00
10.00
15.00
CCR5
SCN
3A
SCN3AP2X3
0.25 0.5 1 2 4 80.03
1.00
32.00
1024.00
CCR5
P2X
3
P2X4
0 1 2 3 4 50.00
2.00
4.00
6.00
8.00
CCR5
P2X4
Correlation; TNFR1 v Purinergic Receptors Correlation; TNFR1 v Voltage gated sodium channel
P2X3
1 2 4 80.03
1.00
32.00
1024.00
P2X3
TNFR1
P2X4
1 2 4 81.00
2.00
4.00
8.00P2X4
TNFR1
SCN3A
1 2 4 80.50
1.00
2.00
4.00
8.00
16.00
SCN3A
TNFR1
SCN9A
1 2 4 80.120.250.501.002.004.008.0016.00
TNFR1
SCN9A
n = 21, r = 0.036, p = 0.876
n = 21, r = -0.294, p = 0.1958
n = 21, r = 0.546, p = 0.0104
n = 21, r = 0.265, p = 0.245
n = 21, r = -0.145, p = 0.531
n = 21, r = 0.297, p = 0.192
n = 21, r = -0.059, p = 0.799
n = 21, r = -0.13, p = 0.554
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Universitätsklinik für Frauenheilkunde
CCR5 and SCN3A expression in endometriotic associated nerves;
CCR5 +ve CCR5 -ve0
2000
4000
6000
8000
SC
N3A
exp
ress
ion
(Flu
ores
cene
inte
nsity
)
**
Overlay CCR5 SCN3A (Nav1.3)
Nerve cells positive for CCR5 have significantly stronger SCN3A expression
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Universitätsklinik für Frauenheilkunde
Increased Nav1.3 expression in CCR positive endometriosis-associated nerves
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Universitätsklinik für Frauenheilkunde
Conclusions
• CCR5, TNFR1, P2X4, SCN3A and SCN9A are increased in ectopic endometriosis
• CCR5 and SCN3A expression are related and co-localised
• Positive CCR5 expression results in increased SCN3A expression
RANTES- through interaction with CCR5 receptor may stimulate an increase in SCN3A expression – possible mechanism in increased pain transmission
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Universitätsklinik für Frauenheilkunde
Acknowledgements
Endometriosis and Gynaecological Oncology Laboratory • Prof Nick Bersinger • Dr Vida Kocbek • Anne Vaucher
Department of Gynaecology, Frauenklinik, Bern
• Prof Michael Müller • Dr Kostantinos Nirgianakis • Dr Sara Imboden
Live cell imaging Laboratory • Dr Fabian Blank • Carlos Wotzkow