Development of skeleton in mammals is an exceedingly complex process. Completion of both endochondral and intramembranous ossification entails a highly intricate but well-coordinated process of patterning, cell fate commitment, differentiation, growth, and remodeling. These events are specified by a coordinated temporal and spatial pattern of gene expression. At first, secreted morphogens such as hedgehog, bone morphogenetic proteins, wingless proteins, and others, signal to key transcription factors to specify gene expression. Runx2 is an essential transcription factor for both chondrocyte and osteoblast differentiation. Runx2 gene deletion results in embryonic lethality due to a complete failure of bone formation. In humans, mutation of the Runx2 gene causes cleidocranial dysplasia, a dominantly inherited skeletal disorder. Another master regulator of skeletogenesis is the Specificity protein-7 (Sp7). Sp7 belongs to the Sp subgroup of the Krppel-like family of transcription factors characterized by three zinc-finger DNA- binding domains. Deletion of Sp7 gene results in failure of osteoblasts, and bone formation. In humans, mutation of the Sp7 gene is linked with the recessive form of osteogenesis imperfecta, skeletal fragility and delayed tooth eruption. However, very little is known about the underlying molecular mechanism for the surprisingly similar phenotype from the two seemingly unrelated proteins. Runx2 is required for the expression of Sp7, as mice with targeted disruption of the Runx2 gene completely lack expression of Sp7. In sharp contrast, the Runx2 expression is normal in the skeletal cells of Sp7 null animals. The functional incompetency of Runx2 in the Sp7 null mice suggests that Sp7 presence is obligatory for completion of the Runx2 osteogenic activity. It is important to note that the observation of Runx2 expression in Sp7 null mice is limited to only RNA, determined by in situ hybridization of embryonic tissues. Our data show that despite normal levels of Runx2 mRNA, Runx2 protein is highly unstable in skeletal tissues of Sp7 null mice. We further demonstrate that Runx2 and Sp7 proteins form a molecular complex and their transcriptional activity is regulated by unique posttranslational modifications. Our findings strongly suggest that in skeletal cells, Sp7 acts as a molecular rheostat and is necessary for functional stability and turnover of Runx2 protein. Our experiment will assess endogenous levels of Runx2 protein in Sp7 null background and by a regulated and selective gene reconstitution in osteoprogenitor cells. The goal of this application is to identify and define a) spatial and temporal organization and assembly of Runx2 and Sp7 regulatory complexes for the formation and/or maintenance of osteoblasts and b) mechanisms supporting the stable complex formation and retention of competency for skeletal gene expression. Knowledge obtained from this study will provide molecular insights into components of a bone regulatory complex that can be targeted for innovative therapy to improve cartilage and bone formation and repair.
Crucial understanding of the molecular mechanism involved in the regulation of bone cell maturation has significant potential for developing interventional therapies in growth anomalies and metabolic bone disorders. Findings from this study will help us in understanding the pathophysiology of skeletal tissues and cartilage and bone disorders.
|Bae, Ji-Myung; Clarke, John C; Rashid, Harunur et al. (2018) Specificity Protein 7 Is Required for Proliferation and Differentiation of Ameloblasts and Odontoblasts. J Bone Miner Res 33:1126-1140|
|Rashid, Harunur; Chen, Haiyan; Hassan, Quamarul et al. (2017) Dwarfism in homozygous Agc1CreERT mice is associated with decreased expression of aggrecan. Genesis 55:|
|Martinez, Milka; Hinojosa, Marcela; Trombly, Daniel et al. (2016) Transcriptional Auto-Regulation of RUNX1 P1 Promoter. PLoS One 11:e0149119|
|Li, Juan; Pan, Qianying; Rowan, Patrick D et al. (2016) Heparanase promotes myeloma progression by inducing mesenchymal features and motility of myeloma cells. Oncotarget 7:11299-309|
|Trotter, Timothy N; Li, Mei; Pan, Qianying et al. (2015) Myeloma cell-derived Runx2 promotes myeloma progression in bone. Blood 125:3598-608|
|Heair, Hannah M; Kemper, Austin G; Roy, Bhaskar et al. (2015) MicroRNA 665 Regulates Dentinogenesis through MicroRNA-Mediated Silencing and Epigenetic Mechanisms. Mol Cell Biol 35:3116-30|
|Adhami, Mitra D; Rashid, Harunur; Chen, Haiyan et al. (2015) Loss of Runx2 in committed osteoblasts impairs postnatal skeletogenesis. J Bone Miner Res 30:71-82|
|Adhami, Mitra D; Rashid, Harunur; Chen, Haiyan et al. (2014) Runx2 activity in committed osteoblasts is not essential for embryonic skeletogenesis. Connect Tissue Res 55 Suppl 1:102-6|
|Rashid, Harunur; Ma, Changyan; Chen, Haiyan et al. (2014) Sp7 and Runx2 molecular complex synergistically regulate expression of target genes. Connect Tissue Res 55 Suppl 1:83-7|
|Chen, Haiyan; Ghori-Javed, Farah Y; Rashid, Harunur et al. (2014) Runx2 regulates endochondral ossification through control of chondrocyte proliferation and differentiation. J Bone Miner Res 29:2653-65|
Showing the most recent 10 out of 14 publications