Abstract 4972: Putative mechanism of TAP63α regulation of chondrocyte maturation during long bone development

Abstract

Abstract p63 is the mouse homologue of the well-known tumor suppressor P53. While it remains controversial as to whether P63 functions as a tumor suppressor gene or an oncogene, mouse genetic studies and mutation analysis of human SHFM syndrome (Split Hand/Foot Malformation) have clearly demonstrated that P63 plays an important role in skin and limb development. The truncated limbs observed in p63 null mice suggest that P63 may contribute to long bone development by playing a role during endochondral bone formation. However, how P63 regulates target gene expression and is linked to endochondral ossification remains largely unknown. Here we present some preliminary data suggesting that P63 may regulate type X collagen gene (Col10a1) expression, thereby impacting the process of chondrocyte maturation. The type X collagen gene (Col10a1) is specifically expressed in hypertrophic chondrocytes, a critical cell stage linking both skeletal development and multiple skeletal diseases. We have previously shown that a 150-bp Col10a1 cis-enhancer is sufficient to mediate its cell-specific expression in vivo. Our further in vitro studies suggest that Runx2, a master transcription factor for osteoblast differentiation as well as for chondrocyte maturation, is one of the major factors that regulate Col10a1 expression via interaction with its tandem repeat Runx2 binding sites. Interestingly, by yeast one-hybrid approach using this cis-enhancer as bait, we identified p53 related proteins as candidate factors that may contribute to the regulation of Col10a1 expression. We have performed Real-time RT-PCR to examine p63 and Runx2 expression in MCT cells, a cell model that shows significant upregulation of Col10a1 upon growth arrest. Both p63 and Runx2 showed around 2-fold upregulation in hypertrophic MCT cells. These data suggest that P63 may collaborate with Runx2 and together regulate Col10a1 expression during chondrocyte maturation. To further explore the putative function of P63 during skeletal development, we have successfully established transgenic mouse lines in which HA and Flag tagged human TAP63α cDNA (the longest P63 isotype) is driven by previously described hypertrophic chondrocyte-specific Col10a1 control element (Col10a1-TAP63α). By phenotypic analysis of these transgenic mice during different developmental stages, we will be able to define the consequence and the potential mechanism of P63 on skeletal development and skeletal disorders. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4972.