Estrogens are important for the maintenance of bone mineral density and the prevention of osteoporosis (or low bone mass), a disease that affects more than 29.5 million people in the United States. Estrogen receptor alpha (ER?), in response to estrogens, increases bone mineral density by increasing osteoblast proliferation and differentiation. GATA4, as a pioneer factor (a protein that can bind to target DNA within silent chromatin and initiate chromatin remodeling and helps recruit other transcription factors to the same locus), helps determine these tissue-specific effects of ER? gene regulation in osteoblasts. However, when and where GATA4 binds to DNA, and how GATA4 directs ER? binding to enhancers to regulate osteoblast differentiation is currently unknown. The central hypothesis of this proposal is that GATA4 increases differentiation of mesenchymal stem cells (MSCs) to osteoblasts by opening chromatin for ER? binding to enhancers. This hypothesis will be tested with the following three specific aims.
Aim 1 will determine when during differentiation ER? and GATA4 are necessary to regulate osteoblast differentiation using temporally regulated gene expression in vitro and in vivo.
Aim 2 will identify where ER? and GATA4 bind to DNA to regulate osteoblast differentiation using state-of-the-art high throughput, whole-genome sequencing technology.
Aim 3 will elucidate the mechanism by which GATA4 regulates ER? binding to osteoblast enhancers. Together these experiments will for the first time describe how the novel osteoblast transcription factor GATA4 regulates estrogen biology in bone. These results will have valuable impact through characterization of ER? tissue-specificity in bone and hence will further understanding of how estrogens prevent osteoporosis.

Public Health Relevance

Postmenopausal osteoporosis or low bone mass affects over 44 million American women and estimates indicate 1 out of 2 women over the age of 50 will experience an osteoporotic fracture in their lifetime. Estrogens are important in the development of bone and maintenance of bone mineral density. The goal of this application is to further develop our functional and mechanistic understanding of how the protein GATA4 controls estrogen biology specifically in bone cells.

National Institute of Health (NIH)
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Research Project (R01)
Project #
Application #
Study Section
Molecular and Cellular Endocrinology Study Section (MCE)
Program Officer
Nicks, Kristy
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Tennessee Health Science Center
Schools of Medicine
United States
Zip Code
Khalid, Aysha B; Slayden, Alexandria V; Kumpati, Jerusha et al. (2018) GATA4 represses RANKL in osteoblasts via multiple long-range enhancers to regulate osteoclast differentiation. Bone 116:78-86
Khalid, Aysha B; Slayden, Alexandria V; Kumpati, Jerusha et al. (2018) GATA4 Directly Regulates Runx2 Expression and Osteoblast Differentiation. JBMR Plus 2:81-91
Martin, Anthony; Yu, Jiali; Xiong, Jian et al. (2017) Estrogens and androgens inhibit association of RANKL with the pre-osteoblast membrane through post-translational mechanisms. J Cell Physiol 232:3798-3807
Morimoto, Eri; Li, Meng; Khalid, Aysha B et al. (2017) Glucocorticoids Hijack Runx2 to Stimulate Wif1 for Suppression of Osteoblast Growth and Differentiation. J Cell Physiol 232:145-53
Khalid, Aysha B; Krum, Susan A (2016) Estrogen receptors alpha and beta in bone. Bone 87:130-5
Martin, Anthony; Xiong, Jian; Koromila, Theodora et al. (2015) Estrogens antagonize RUNX2-mediated osteoblast-driven osteoclastogenesis through regulating RANKL membrane association. Bone 75:96-104