Testosterone administration increases skeletal muscle mass and strength, and decreases fat mass in men, but the mechanisms by which testosterone regulates body composition are poorly understood. In the previous funding cycle, we demonstrated that testosterone and DHT promote the differentiation of adult, multipotent. mesenchymal stem cells into myogenic lineage and inhibit their differentiation into adipogenic lineage through an androgen receptor (AR) mediated mechanism. We also discovered that androgens modulate the differentiation of mesenchymal, multipotent cells by activation of Wnt signaling through the association of liganded AR to cytosolic catenin, which causes the latter to translocate to the nucleus and activate Wnt- target genes that in turn regulate mesenchymal multipotent cell differentiation. In this competing continuation application, our objective is to elucidate downstream signaling mechanisms by which AR: b-catenin interaction regulates mesenchymal stem cell differentiation. Our preliminary data show that testosterone upregulates a number of Wnt-target genes, including follistatin. Follistatin inhibits the activity of a number of TGF-b family members, including myostatin, which inhibit myogenesis. We hypothesize that testosterone modulates myogenic differentiation by activation of follistatin, resulting in the inhibition of TGF-/SMAD signaling. We will utilize both in vitro and in vivo approaches to test these hypotheses.
Aim 1 will determine whether follistatin is essential for mediating the effects of testosterone on mesenchymal multipotent cell differentiation. We will determine whether knockdown of follistatin through siRNAs blocks testosterone's action on myogenic differentiation in vitro. We will confirm in vivo whether genetic disruption of follistatin expression in Cre/Lox mice blocks testosterone's effects on muscle mass.
Aim 2 will determine whether testosterone inhibits TGF/SMAD signaling by its effects on follistatin. We will investigate whether testosterone blocks SMAD signaling by up regulating SMAD7 and by inhibiting SMAD 3 phosphorylation and signaling.
Aim 3 (Translational Aim) will determine whether follistatin, acting downstream of AR and b catenin, will provide greater selectivity, and improve muscle mass and function without affecting prostate growth. These translational studies would unveil the mechanisms by which androgens regulate body composition and provide a rational framework for the proposed application of androgens, SARMs, and follistatin as function-promoting therapies. Follistatin, being downstream of AR and b-catenin, would be a promising function promoting anabolic molecule that would increase skeletal muscle mass and function without affecting prostate growth in older men, and in men with prostate cancer who are receiving androgen deprivation therapy. Public Health Relevance: Aging-associated limitations of physical function impose an enormous health and socioeconomic burden on the individual and the society;therefore, there is substantial unmet need for the development of function promoting anabolic therapies. The proposed translational studies will elucidate the signaling mechanisms by which testosterone increases skeletal muscle mass and provide novel leads for the discovery of more selective anabolic molecules, such as follistatin, that might increase skeletal muscle mass and function without affecting the prostate. The proposed project also provides an outstanding opportunity to further our understanding of hormonal regulation of stem cell differentiation and cell fate.

Public Health Relevance

Aging-associated limitations of physical function impose an enormous health and socioeconomic burden on the individual and the society;therefore, there is substantial unmet need for the development of function promoting anabolic therapies. The proposed translational studies will elucidate the signaling mechanisms by which testosterone increases skeletal muscle mass and provide novel leads for the discovery of more selective anabolic molecules, such as follistatin, that might increase skeletal muscle mass and function without affecting the prostate. The proposed project also provides an outstanding opportunity to further our understanding of hormonal regulation of stem cell differentiation and cell fate.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
7R01AG037193-10
Application #
8652546
Study Section
Aging Systems and Geriatrics Study Section (ASG)
Program Officer
Williams, John
Project Start
2004-09-30
Project End
2015-01-31
Budget Start
2013-06-15
Budget End
2014-01-31
Support Year
10
Fiscal Year
2013
Total Cost
$395,536
Indirect Cost
$171,118
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Xue, Ran; Zakharov, Mikhail N; Xia, Yu et al. (2014) Research resource: EPSLiM: ensemble predictor for short linear motifs in nuclear hormone receptors. Mol Endocrinol 28:768-77
Jasuja, Ravi; Costello, James C; Singh, Rajan et al. (2014) Combined administration of testosterone plus an ornithine decarboxylase inhibitor as a selective prostate-sparing anabolic therapy. Aging Cell 13:303-10
Serra, Carlo; Sandor, Nicolae Lucian; Jang, Hyeran et al. (2013) The effects of testosterone deprivation and supplementation on proteasomal and autophagy activity in the skeletal muscle of the male mouse: differential effects on high-androgen responder and low-androgen responder muscle groups. Endocrinology 154:4594-606
Guo, Wen; Bachman, Eric; Li, Michelle et al. (2013) Testosterone administration inhibits hepcidin transcription and is associated with increased iron incorporation into red blood cells. Aging Cell 12:280-91
Kumar, R; Atamna, H; Zakharov, M N et al. (2011) Role of the androgen receptor CAG repeat polymorphism in prostate cancer, and spinal and bulbar muscular atrophy. Life Sci 88:565-71
Serra, Carlo; Bhasin, Shalender; Tangherlini, Frances et al. (2011) The role of GH and IGF-I in mediating anabolic effects of testosterone on androgen-responsive muscle. Endocrinology 152:193-206
Zakharov, Mikhail N; Pillai, Biju K; Bhasin, Shalender et al. (2011) Dynamics of coregulator-induced conformational perturbations in androgen receptor ligand binding domain. Mol Cell Endocrinol 341:1-8
Zakharov, M N; Ulloor, J; Bhasin, S et al. (2011) Guanidinium chloride-induced spectral perturbations of 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid confound interpretation of data on molten globule states. Anal Biochem 416:126-8
Lee, Se-Jin; Lee, Yun-Sil; Zimmers, Teresa A et al. (2010) Regulation of muscle mass by follistatin and activins. Mol Endocrinol 24:1998-2008
Flanagan, John N; Linder, Kristina; Mejhert, Niklas et al. (2009) Role of follistatin in promoting adipogenesis in women. J Clin Endocrinol Metab 94:3003-9