The Impaired Bone Anabolic Effect of PTH in the Absence of Endogenous FGF2 is Partially Due to Reduced ATF4 Expression

Parathyroid hormone (PTH) is currently the only approved anabolic agent for osteoporosis pharmacotherapy in the USA. However, the molecular and cellular mechanisms underlying which intermittent PTH stimulates bone formation are not fully established. Activating transcription factor 4 (ATF4) was recently identified to be a downstream target of PTH signaling in osteoblasts and FGF2 is able to rapidly increase ATF4 mRNA and protein expression in osteoblasts. Furthermore, ATF4 expression is markedly reduced in Fgf2−/− osteoblasts. In addition, FGF2 is required for the anabolic action of PTH on bone formation. Therefore, we hypothesize that the impaired anabolic effect of PTH in Fgf2−/− mice is partially due to reduced ATF4 expression. To test this hypothesis, we examined the ability of PTH to increase ATF4 expression in vitro and in vivo. In vitro data showed that PTH induced a significant increase in ATF4 mRNA expression as early as 15 min in Fgf2+/+ primary bone marrow stromal cells (BMSCs) but not in Fgf2−/− BMSCs. In vivo data showed that treatment with PTH (1–34) (40 μg/kg/d) treatment for 2 weeks in 21–23 months female mice increased lumbar vertebrae bone mineral density in Fgf2+/+ (13.8% increase). In contrast there was a 2.1% decrease in Fgf2−/− mice. Interestingly, basal ATF4 mRNA expression in tibiae was significantly lower in Fgf2−/− mice (46% decrease) compared to Fgf2+/+ mice. PTH treatment increased ATF4 mRNA by 97% (p\u3c 0.05) in Fgf2+/+ compared to 8% (p = 0.57) in Fgf2−/− mice. Immunohistochemistry of vertebrae showed less ATF4 staining in Fgf2−/− tissue, and treatment with PTH increased ATF4 staining in Fgf2+/+ but the increase was attenuated in Fgf2−/− tissue. In summary, reduced ATF4 expression may result in decreased osteoblast differentiation, and possibly contribute to the impaired stimulation of PTH on bone formation in Fgf2−/− mice