Fontaine C. *, Adlanmerini M. *, Buscato M. *, Smirnova N *, Lupieri A. *, Vinel A. *, Guillaume M., * Zahreddine R. *, Knauf C. *, Kim SH.
†, Katzenellenbogen BS.
‡, Katzenellenbogen JA.
†, Laffargue M *, Gourdy P.*, Valera MC.*, Lenfant F. *, Arnal JF*
* Université de Toulouse 3, Institut National de la Santé et de la Recherche Médicale (INSERM) UMR1048, Institut des Maladies Métaboliques et Cardiovasculaires, Toulouse, France ; † Department of Molecular and Integrative Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois, United States of America ‡ Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois , United States of America
Vascular effects of ER α αα α membrane initiated signaling
Background
Genomic effect
Membrane Initiated steroid Signaling
Palmitoylation
OH
OH H
H H
H2
CNH
PAMAM NH2NH2
NH2
NH2 NH2 NH2 NH2 H2 N (
)~ 20
( )~ 236
Chamblisset al. J Clin Invest 2014 Estrogen Dendrimer
Conjugated (EDC)
OH
OH H
H H
OH
OH
Estetrol (E4)
Abot et al. EMBO Mol Med 2014
ERαAF20
ERαC451A Adlanmeriniet al. PNAS 2014
Billon-Gales et al. PNAS 2011
Zα αα αD/^^ ŝƐŶĞĐĞƐƐĂƌLJĂŶĚƐƵĨĨŝĐŝĞŶƚƚŽŝŶĚƵĐĞ ĞŶĚŽƚŚĞůŝĂůŚĞĂůŝŶŐ
Zα αα αD/^^ ŝƐŶĞĐĞƐƐĂƌLJƚŽŝŶĐƌĞĂƐĞEKƉƌŽĚƵĐƚŝŽŶ ŝŶƌĞƐƉŽŶƐĞƚŽϮ
ŶĚŽƚŚĞůŝĂů ĐĞůů ŵŽƌƉŚŽůŽŐLJ ŝƐ ĂůƚĞƌĞĚ ŝŶZα αα αϰϱϭŵŝĐĞ
KZd/E
<Z^ ͲZ/E'Z ϯϳ Σ
WZK E2
Recording of electric current generated by oxidation of NO
Validation of the C451A-ERĮ mouse model
72h
Quantification of reendothelialization weeks
0 4 6 8
Carotid arteries
Aorta
0 250
500 WT DMSO
WT E2 C451A-ERαDMSO C451A-ERαE2
NO release from basal (nM)
Adlanmeriniet al. PNAS 2014
6 28 days Treatment (E2, EDC or E4)
10 4
3
Zα αα αD/^^ ŝƐĚŝƐƉĞŶƐĂďůĞĨŽƌƚŚĞƉƌĞǀĞŶƚŝŽŶ
ŽĨĞĂƌůLJĂƚŚĞƌŽŵĂĂƚƚŚĞĂŽƌƚŝĐƐŝŶƵƐŝŶƚŚĞŵŽƵƐĞ
Vehicle E41 1.5 2
ĐƚŝǀĂƚŝŽŶŽĨŶƵĐůĞĂƌ͕ďƵƚŶŽƚŵĞŵďƌĂŶĞ͕Zɲ ŝƐ ƐƵĨĨŝĐŝĞŶƚĨŽƌƚŚĞƐƵƉƉƌĞƐƐŝŽŶŽĨŶĞŽŝŶƚŝŵĂů
ŚLJƉĞƌƉůĂƐŝĂ
Empty Dendrimer
Neointima/Media Ratio E2
0.5 1 1.5 2
Vehicle EDC
Vehicle
EDC Sinus Thoracic aorta Greater
Curvature Lower Curvature
*
§§§
(*P génotype<0.05; p aortic area<0.001)
§§§
§§§
ERĮ-C451A
Thoracic aorta ERĮWT
50 µm 50 µm
months 5 Intact mice
Surface (µm2)
Hypercholesterolemic diet
0.0 0.5 1.0 1.5
**
Vehicle E2
50 150 250 350 450 550 Time (seconds)
Delta N
Adlanmeriniet al. PNAS 2014
***
0.5
Vehicle E4
Vehicle E2 weeks
4 6 18
Hypercholesterolemic diet (Cholesterol 1.25%) ERĮ-C451A LDLr-/-
ERĮ-WT LDLr-/- ERĮ-C451A LDLr-/- 50
100 150 200 E2
ERα-WT LDLr-/-ERα-C451A LDLr-/- Vehicle
Lesionarea x103µm²
VCAM 1
0.5 1.0 1.5 2.0 2.5
relative mRNAlevel
PinteractionNS, PE2***, PgenotypeNS
MCP1
0.5 1.0 1.5
Gremlin 2
2 4 6 8 10
Pinteraction: 0.95, PE2 = 0.006 **, P genotypeNS
Fatty streak
PinteractionNS, PE2***, PgenotypeNS
PinteractionNS, PE2***, PgenotypeNS
KE>h^/KE
weeks
4 6 12
yp (Cholesterol1.25%)
Plasma Cholesterol
Vehicle E2 Dendrimer EDC 0
500 1000
mg/dl *
Dendrimer EDC
Vehicle E2
Vehicle E2 DendrimerEDC 0
20 40 60 80
Lesionsaera(x103µm²)
***
Fatty streak LDLr-/-
ZɲD/^^ ĂƉƉĞĂƌƐƚŽďĞŶĞĐĞƐƐĂƌLJƚŽƐƵƉƉŽƌƚ ƐŽŵĞ͕ďƵƚŶŽƚĂůů͕ǀĂƐĐƵůĂƌƉƌŽƚĞĐƚŝǀĞĂĐƚŝŽŶƐŽĨ
ĞƐƚƌŽŐĞŶ͘
Smirnovaet al. Circ. Res. 2015
Genetically modified mouse model
= Loss-of function
Pharmacological approaches
=Gain of function
'ĞŶŽƚLJƉĞ ERααααAF20 (nuclear)
ERαααα-C451A (MISS)
td
dƌĞĂƚŵĞŶƚ ƐƚƌĂĚŝŽů Estetrol
(nuclear) dĂŵŽdžŝĨĞŶ
;&ϭͿ
;D/^^Ϳ
Acceleration of endothelial healing
YES 1 NO 2 NO 3 NO 4 YES 2
Increase of NO production
nd NO 2 NO 3 nd YES 2
Prevention of neointimal hyperplasia
nd nd YES 5 YES 5 NO 5
Prevention of atheroma at aortic sinus
NO 1 YES YES 3 YES 4 NO
Summarize of the different studies describing the respective role of nuclear ERααααand ERααααMISS in vasculoprotection using genetically modified mouse models (loss-of-function) and pharmacological tools (gain-of-function).
Nd = not determined
1. Billon-Gales A et al. Proc Natl Acad Sci U S A. 2011;108:13311-13316 2. Adlanmerini M, et al. Proc Natl Acad Sci U S A. 2014;111:E283-290 3. Abot A et al EMBO Molecular Medicine2014;6:1328-1346 4. Fontaine C et al. Am J Pathol. 2013;183:304-312 5. Smirnova NF et al. Circulation Research2015
