Both androgens and estrogens contribute to the maintenance of bone mass during adult life in men, but the cell targets and molecular mechanisms of these effects remain poorly understood. During the preceding funding period, we found that the effects of androgens on cancellous bone result from AR signaling in cells of the mesenchymal lineage leading to a decrease in osteoclasts; and orchidectomy (ORX) increases soluble RANKL levels as well as the number of B-lymphocytes in the murine bone marrow. RANKL derived from osteocytes is critical for cancellous bone remodeling and B-lymphocyte-derived RANKL contributes to the loss of cancellous bone mass in ovariectomized (OVX) mice. In cortical bone, however, the effects of androgens do not require AR signaling in any cell type of the mesenchymal lineage, nor do they require ER? signaling downstream of committed osteoblast progenitors targeted by Osx1-Cre, or AR or ER? signaling in the osteoclast lineage. Estrogens, nonetheless, attenuate endocortical resorption in females by ER? signaling in uncommitted mesenchymal progenitors targeted by Prx1-Cre. In addition, the mRNA and secreted protein levels of SDF1 ? a chemotactic cytokine of the CXC family that is abundantly expressed in Prx1-Cre targeted cells and promotes osteoclast generation ? are higher in GFP-tagged ER? null cells as well as bone marrow cell cultures from female ER?f/f;Prx1-Cre mice as compared to identical cultures from littermate controls; and so is osteoclastogenesis in co-cultures of ER? null BM stromal or calvaria cells with macrophages. Furthermore, both OVX and ORX increase SDF1 in the bone marrow plasma, administration of E2 to ORX mice prevents this increase, and E2, but not DHT, prevents cortical bone loss in ORX mice. Finally, we found that the OVX- or ORX-induced cortical bone loss is prevented by restraining H2O2 generation in osteoclast mitochondria; and the loss of cortical bone mass in OVX mice is prevented by a 17?-E2 dendrimer conjugate (EDC), incapable of stimulating nuclear- initiated actions of ER?. Based on these advances, we will test the interrelated hypotheses that: in males, the protective effects of androgens on cancellous bone result from AR signaling in osteocytes, B-lymphocytes, or both. These actions result in the attenuation of RANKL and thereby attenuation of the number of cancellous osteoclasts. The protective effects of androgens against endocortical resorption result, at least in part, from ER?- mediated actions (upon aromatization of androgens to estrogens) on Prx1-Cre targeted uncommitted mesenchymal progenitors. These actions repress SDF1 production, thereby attenuating osteoclast formation and homing at the endosteal surfaces. The suppressive effect of estrogens on SDF1 production ultimately leads to the restraining of H2O2 accumulation in osteoclasts and results from non-nuclear-initiated signaling of the ER?. To advance these hypotheses we will try to elucidate the mechanism of the protective effects of androgens on cancellous bone by determining the role of osteocyte- and B-lymphocyte-derived RANKL as well as the role of the B lymphocyte AR in these effects. To do this we will study: i) mice in which RANKL is deleted from mature osteoblasts/osteocytes (RANKLf/f;Dmp1-Cre), ii) mice in which both AR and RANKL are deleted from osteoblasts/osteocytes (RANKLf/f;ARf/y;Dmp1-Cre), iii) mice in which RANKL is deleted from B-lymphocytes (RANKLf/f;CD19-Cre), and iv) mice in which AR is deleted from B-lymphocytes (ARf/y;CD19-Cre). In addition, we will elucidate the mechanism of the protective effects of androgens on cortical bone by investigating: a) whether down-regulation of SDF1 in uncommited mesenchymal progenitors is responsible for some of these effects using: i) mice in which SDF1 is deleted in Prx1-Cre targeted cells, and ii) mice in which the SDF1 receptor, CXCR4, is deleted in LysM-Cre targeted cells; and b) whether the effect of androgens results from ER?-mediated actions upon androgen aromatization to estrogens using ER?f/f;Prx1-Cre mice. Using co-cultures of stromal and hematopoietic cells, we will also examine whether SDF1 promotes osteoclastogenesis via H2O2; and whether the effect of estrogens on SDF1 results from non-nuclear-initiated signaling of the ER?.
Men produce both androgens and estrogens and both of these hormones help to prevent the development of osteoporosis and the related fractures. However, it is not clear exactly how these hormones work and how much of their protective role is due to androgens versus estrogens. Studies of the applicant with mouse models during the last twenty five years have provided some important clues. The research proposed in this grant application is a logical extension of the applicant?s work on the subject and aims to provide novel insights, not only into how these hormones act on bone but also to discover new therapies for the management of osteoporosis in several conditions, such as male veterans with prostate cancer receiving drugs that block the production of androgens or female veterans with breast cancer who take drugs that block the production of estrogens. Such therapies could prevent the development of osteoporosis in both women and men without unwanted effects on the male and female reproductive organs.
|Weinstein, Robert S; Hogan, Erin A; Borrelli, Michael J et al. (2017) The Pathophysiological Sequence of Glucocorticoid-Induced Osteonecrosis of the Femoral Head in Male Mice. Endocrinology 158:3817-3831|
|Ucer, Serra; Iyer, Srividhya; Kim, Ha-Neui et al. (2017) The Effects of Aging and Sex Steroid Deficiency on the Murine Skeleton Are Independent and Mechanistically Distinct. J Bone Miner Res 32:560-574|
|Almeida, Maria; Laurent, Michaël R; Dubois, Vanessa et al. (2017) Estrogens and Androgens in Skeletal Physiology and Pathophysiology. Physiol Rev 97:135-187|
|Iyer, Srividhya; Han, Li; Ambrogini, Elena et al. (2017) Deletion of FoxO1, 3, and 4 in Osteoblast Progenitors Attenuates the Loss of Cancellous Bone Mass in a Mouse Model of Type 1 Diabetes. J Bone Miner Res 32:60-69|
|Carson, James A; Manolagas, Stavros C (2015) Effects of sex steroids on bones and muscles: Similarities, parallels, and putative interactions in health and disease. Bone 80:67-78|
|Almeida, Maria; Iyer, Srividhya; Martin-Millan, Marta et al. (2013) Estrogen receptor-? signaling in osteoblast progenitors stimulates cortical bone accrual. J Clin Invest 123:394-404|
|Manolagas, Stavros C; O'Brien, Charles A; Almeida, Maria (2013) The role of estrogen and androgen receptors in bone health and disease. Nat Rev Endocrinol 9:699-712|
|Iyer, Srividhya; Ambrogini, Elena; Bartell, Shoshana M et al. (2013) FOXOs attenuate bone formation by suppressing Wnt signaling. J Clin Invest 123:3409-19|
|Manolagas, Stavros C; Cummings, Steven R (2013) Skeletal aging: from bench to bed side. J Gerontol A Biol Sci Med Sci 68:1195-6|
|Andrade, Anenisia C; Baron, Jeffrey; Manolagas, Stavros C et al. (2010) Hormones and genes of importance in bone physiology and their influence on bone mineralization and growth in Turner syndrome. Horm Res Paediatr 73:161-5|