Effects of recombinant Irisin on the musculoskeletal system of hind-limb suspended mice

We previously showed that Irisin, a myokine released from skeletal muscle after physical exercise, plays a central role in the control of bone mass, driving positive effects on cortical mineral density and geometry in vivo (1). Here we demonstrated that r-Irisin treatment prevents bone loss in hind-limb suspended mice when administered during suspension and recovers bone mass when mice were injected after a suspension period (4 weeks) during which they developed bone loss. Micro computed tomography of femurs showed that r-Irisin treatment positively affected both cortical and trabecular bone. As expected, unloaded mice treated with vehicle displayed a remarkable decrease of cortical and trabecular bone mineral density (BMD), whereas in Irisin-treated unloaded mice no loss of BMD was observed with respect to control mice kept under normal loading. Likewise, by treating mice after they already developed disuse-induced bone loss, r-Irisin was able to restore the damaged mineral component. Furthermore, trabecular bone volume fraction (BV/TV), which dramatically decreased in unloaded mice, was prevented by r-Irisin therapy. In particular, r-Irisin treatment preserved the number of trabeculae (Tb.N) and the fractal dimension, an index of optimal micro-architectural complexity of trabecular bone.We also showed that r-Irisin treatment protects muscle mass suffering from atrophy during unloading. Thus, unloaded mice treated with vehicle displayed a severe loss of muscle mass, as confirmed by ~ 60% decline of vastus lateralis weight and ~33% decrease of fiber cross-sectional area. Conversely, Irisin-treated unloaded mice showed no loss of muscle weight and similar fiber cross-sectional area to control mice. Our data reveal for the first time that r-Irisin treatment prevents and retrieves disuse-induced bone loss and muscle atrophy. These findings may lead to develop an Irisin-based therapy for the prevention and treatment of osteoporosis and sarcopenia in all patients who cannot perform physical activity, as occurs during aging and immobility, and it could also represent a countermeasure for astronauts exposed to microgravity during space flight missions.This work was supported in part by ERISTO grant (to M.G.), by MIUR grant ex60% (to M.G.) and by SIOMMMS grant (to G.C.)

Irisin injected mice display increased tibial cortical mineral density and polar moment of inertia

It has been recently reported that, after physical activity, the skeletal muscle releases Irisin, the newly identified myokine able of driving transition of white adipocytes into brown [1]. This result supported the role of skeletal muscle as endocrine organ, suggesting that it could target other tissues besides adipose tissue. In our previous work, we demonstrated that conditioned media collected from primary myoblasts of exercised mice were able to increase OB differentiation and this effect was Irisinmediated [2]. Here we show that Irisin has positive effect on cortical mineral density and geometry in vivo. Young male mice were injected with r-Irisin and cortical bone adaptation was analyzed by micro-CT at tibial midshaft. Our results show that cortical tissue mineral density is significantly increased in Irisin-injected mice compared to vehicle-injected littermates (+7.15%; p<0.01). Furthermore, this higher density of calcium hydroxyapatite at cortical site is accompanied by increase in periosteal circumference (+7.5%; p<0.03) and polar moment of Inertia (pMOI +19,21%; p<0,01). A greater pMOI indicates stronger resistance of a long bone to torsion and, together with higher bone mineral density, suggests higher protection against fracture. The effect of Irisin is fully comparable to the effect of physical activity that is widely accepted method for increasing bone mineral density and bone size in healthy populations. In view of further proving the involvement of Irisin in bone metabolism, we validate its direct effect on osteoblasts in vitro. Phosphorylation of the MAP kinase ERK and the expression of Atf4 were significantly increased after Irisin treatment, as well as ALP and pro-Collagen I mRNA expression. Our data highlight a novel link in muscle-fat-bone axis demonstrating that Irisin targets bone tissue directly, driving positive effects on cortical mineral density and geometry in vivo. These findings would expand the research of exercise-mimetic drugs that might be widely used to treat osteoporotic patients who are suffering from immobilization and cannot perform physical activity

FSH and TSH in the Regulation of Bone Mass: The Pituitary/Immune/Bone Axis

Recent evidences have highlighted that the pituitary hormones have profound effects on bone, so that the pituitary-bone axis is now becoming an important issue in the skeletal biology. Here, we discuss the topical evidence about the dysfunction of the pituitary-bone axis that leads to osteoporotic bone loss. We will explore the context of FSH and TSH hormones arguing their direct or indirect role in bone loss. In addition, we will focus on the knowledge that both FSH and TSH have influence on proinflammatory and proosteoclastogenic cytokine expression, such as TNFα and IL-1, underlining the correlation of pituitary-bone axis to the immune system

Irisin for care and prevention of osteoporosis

An object of the present invention is the use of Irisin for the treatment and/or prevention of osteoporosis. In particular, the present invention refers to the use of recombinant irisin for the treatment and/or prevention of osteoporosis. A method for preventing and/or treating osteoporosis by administering an effective amount of irisin to a subject, is provided

CELLULAR MECHANISMS OF BONE REGENERATION: ROLE OF WNT-1 IN BONE-MUSCLE INTERACTION DURING PHYSICAL ACTIVITY

Wnt1 is one of the several glycoproteins activating Wnt signaling, critical for normal skeletal development and bone homeostasis. Wnt1 was previously believed to solely regulate central nervous system development, in particular in midbrain and cerebellum. However, remarkable findings have recently shown that several patients affected by severe form of Osteogenesis Imperfecta (OI) display a Wnt1 mutation thereby revealing a possible role of Wnt1 in bone metabolism. Here, we show that recombinant Wnt1 (r-Wnt1) strongly increases differentiation of bone marrow stromal cells into mature osteoblasts, as demonstrated by the enhanced number of cells positively stained for alkaline phosphatase, one of the osteoblastic marker genes, whose mRNA levels are also significantly up-regulated. Furthermore, other osteogenic master genes such as Collagen I and Osteopontin are also enhanced when bone marrow precursors were differentiated toward osteoblastic phenotype in the presence of r-Wnt1. Intriguingly, by in vivo and in vitro findings, we report that in the bone marrow of mice subjected to physical activity there is a high endogenous Wnt1 synthesis compared to mice kept in resting conditions. Moreover, conditioned medium collected from ex vivo myoblasts, harvested from exercised mice, up-regulates Wnt1 expression in osteoblast cell cultures obtained from control mice. Overall our findings support the role of Wnt1 in regulating bone metabolism and suggest that this molecule could be one of the mediators through which physical activity may exert beneficial effect on bone

Irisin for care and prevention of osteoporosis

An object of the present invention is the use of Irisin for the treatment and/or prevention of osteoporosis. In particular, the present invention refers to the use of recombinant irisin for the treatment and/or prevention of osteoporosis. A method for preventing and/or treating osteoporosis by administering an effective amount of irisin to a subject, is provided

Irisin enhances osteoblast differentiation in vitro

It has been recently demonstrated that exercise activity increases the expression of the myokine Irisin in skeletal muscle, which is able to drive the transition of white to brown adipocytes, likely following a phenomenon of transdifferentiation. This new evidence supports the idea that muscle can be considered an endocrine organ, given its ability to target adipose tissue by promoting energy expenditure. In accordance with these new findings, we hypothesized that Irisin is directly involved in bone metabolism, demonstrating its ability to increase the differentiation of bone marrow stromal cells into mature osteoblasts. Firstly, we confirmed that myoblasts from mice subjected to 3 weeks of free wheel running increased Irisin expression compared to nonexercised state. The conditioned media (CM) collected from myoblasts of exercised mice induced osteoblast differentiation in vitro to a greater extent than those of mice housed in resting conditions. Furthermore, the differentiated osteoblasts increased alkaline phosphatase and collagen I expression by an Irisin-dependent mechanism. Our results show, for the first time, that Irisin directly targets osteoblasts, enhancing their differentiation. This finding advances notable perspectives in future studies which could satisfy the ongoing research of exercise-mimetic therapies with anabolic action on the skeleton

THE ROLE OF LIGHT ON BONE REMODELING

LIGHT, expressed by different cells of the immune system, binds two trans-membrane receptors: HVEM and LT?R. It is over-expressed in erosive rheumatoid arthritis and lytic myeloma-bone disease and controversial data have been published on its role in vitro osteoclast (OC) formation. Here, we investigated the role of LIGHT on in vitro murine osteoclastogenesis model and bone phenotype in LIGHT-/- mice. Firstly, we showed that murine macrophages stimulated with LIGHT alone did not differentiate into OCs. Consistently, the addition of agonist anti-HVEM and anti-LT?R antibodies did not affect osteoclastogenesis in the same cultures. Interestingly, the presence of LIGHT and sub-optimal RANKL concentration displayed synergic effects on OC formation through the early and sustained activation of Akt, NF?B and JNK pathways. Secondly, by microCT we found that the femurs of LIGHT-/- mice exhibited a decrease in trabecular BV/TV due to a significant reduction in trabecular thickness and number as well as the increase in trabecular spaces respect to wild-type (WT) mice. Further, a deformity of the vertebral column due to the collapse of vertebral bodies was radiographically observed in about 40% of LIGHT-/- mice. To investigate the possible molecular mechanism/s responsible for this bone phenotype in LIGHT-/- mice we studied OPG levels in whole bone marrow extracts from the femurs of these mice and demonstrated a significant reduction in OPG mRNA transcript respect to WT. Based on the knowledge that mature B cells are the major source of OPG and that they represent about 50% of the spleen cells, we treated with LIGHT total splenocytes demonstrating a three fold increase of OPG mRNA levels respect to the untreated cells. In conclusion, our results suggested that the net effect of LIGHT on bone remodeling probably occurs through the modulation of OPG expression. However, further experiments are needed to better clarify this issu

Adrenergic stimulation decreases osteoblast oxytocin synthesis

Oxytocin (OT) regulates bone mass by inducing the differentiation of osteoblasts to a mature, mineralizing phenotype. We have shown recently that osteoblasts can synthesize OT. In view of known interactions between OT-ergic and adrenergic neurons in the central nervous system, we questioned whether the negative regulation of osteoblast differentiation by adrenergic nerves was mediated through its suppression of OT synthesis. We first confirmed that α 1b and β 2 adrenergic receptors were expressed on both primary murine osteoblasts and MC3T3-E1 cells. We then showed that α 1 and β 2 adrenergic agonists downregulated OT synthesis, and that the effect of each agonist was reversed by its respective antagonist. The data suggest that the negative effects of adrenergic stimulation on bone mass could, in part, arise from decreased OT synthesis. © 2011 New York Academy of Sciences