Protective Hematopoietic Effect of Estrogens in a Mouse Model of Thrombosis: Respective Roles of Nuclear Versus Membrane Estrogen Receptor α.
We recently reported that chronic 17β-estradiol (E2) treatment in mice decreases platelet responsiveness, prolongs the tail-bleeding time and protects against acute thromboembolism via the hematopoietic estrogen receptor alpha (ERα), and independently of ERβ. Here, we have explored the respective roles of membrane vs nuclear actions of ERα in this process, using: 1) the selective activator of membrane ERα: estrogen dendrimer conjugate, and 2) mouse models with mutations in ERα. The selective targeting of activation function 2 of ERα provides a model of nuclear ERα loss-of-function, whereas mutation of the ERα palmitoylation site leads to a model of membrane ERα deficiency. The combination of pharmacological and genetic approaches including hematopoietic chimera mice demonstrated that absence of either membrane or nuclear ERα activation in bone marrow does not prevent the prolongation of the tail-bleeding time, suggesting a redundancy of these two functions for this E2 effect. In addition, although hematopoietic membrane ERα is neither sufficient nor necessary to protect E2-treated mice from collagen/epinephrine-induced thromboembolism, the protection against death-induced thromboembolism is significantly reduced in the absence of hematopoietic nuclear ERα activation. Overall, this study emphasizes that hematopoietic cells (likely megakaryocytes and possibly immune cells) constitute an important target in the antithrombotic effects of estrogens, and delineate for the first time in vivo the respective roles of membrane vs nuclear ERα effects, with a prominent role of the latter.
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Marie-Cécile
MC
Valéra
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Coralie
C
Fontaine
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Françoise
F
Lenfant
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Cendrine
C
Cabou
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Maeva
M
Guillaume
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Natalia
N
Smirnova
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Sung Hoon
SH
Kim
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Pierre
P
Chambon
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
John A
JA
Katzenellenbogen
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Benita S
BS
Katzenellenbogen
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Bernard
B
Payrastre
INSERM U1048 and Université Toulouse III I2MC (M.-C.V., C.F., F.L., C.C., M.G., N.S., B.P., J.-F.A.), Toulouse, 31432 France; Faculté de Chirurgie Dentaire (M.-C.V.), Université de Toulouse III, Toulouse, 31432 France; Department of Chemistry (H.H.K., J.A.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institut de Génétique et de Biologie Moléculaire et Cellulaire (P.C.), Collège de France, Université de Strasbourg, Illkirch, 67400 France; Department of Physiology and Cell Biology (B.S.K.), University of Illinois at Urbana-Champaign, Urbana, IL 61801; and Laboratoire d'Hématologie (B.P.), Centre Hospitalier Universitaire de Toulouse, Toulouse, 31432 France.
Jean-François
JF
Arnal
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D004195:Disease Models, Animal; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D004967:Estrogens / Q000627:therapeutic use*; D051379:Mice; D008822:Mice, Transgenic; D013927:Thrombosis / Q000188:drug therapy* / Q000378:metabolism
D047628:Estrogen Receptor alpha; D004967:Estrogens