Clinkenbeard Erica L, Cass Taryn A, Ni Pu, Hum Julia M, Bellido Teresita, Allen Matthew R, White Kenneth E
Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN, USA.
Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN, USA.
J Bone Miner Res. 2016 Jun;31(6):1247-57. doi: 10.1002/jbmr.2792. Epub 2016 Mar 4.
The transgenic and knockout (KO) animals involving Fgf23 have been highly informative in defining novel aspects of mineral metabolism, but are limited by shortened lifespan, inability of spatial/temporal FGF23 control, and infertility of the global KO. To more finely test the role of systemic and genetic influences in FGF23 production, a mouse was developed that carried a floxed ("f")-Fgf23 allele (exon 2 floxed) which demonstrated in vivo recombination when bred to global-Cre transgenic mice (eIIa-cre). Mice homozygous for the recombined allele ("Δ") had undetectable serum intact FGF23, elevated serum phosphate (p < 0.05), and increased kidney Cyp27b1 mRNA (p < 0.05), similar to global Fgf23-KO mice. To isolate cellular FGF23 responses during phosphate challenge, Fgf23(Δ/f) mice were mated with early osteoblast type Iα1 collagen 2.3-kb promoter-cre mice (Col2.3-cre) and the late osteoblast/early osteocyte Dentin matrix protein-1-cre (Dmp1-cre). Fgf23(Δ/f) /Col2.3-cre(+) and Fgf23(Δ/f) /Dmp1-cre(+) exhibited reduced baseline serum intact FGF23 versus controls. After challenge with high-phosphate diet Cre(-) mice had 2.1-fold to 2.5-fold increased serum FGF23 (p < 0.01), but Col2.3-cre(+) mice had no significant increase, and Dmp1-cre(+) mice had only a 37% increase (p < 0.01) despite prevailing hyperphosphatemia in both models. The Fgf23(Δ/f) /Col2.3-cre was bred onto the Hyp (murine X-linked hypophosphatemia [XLH] model) genetic background to test the contribution of osteoblasts and osteocytes to elevated FGF23 and Hyp disease phenotypes. Whereas Hyp mice maintained inappropriately elevated FGF23 considering their marked hypophosphatemia, Hyp/Fgf23(Δ/f) /Col2.3-cre(+) mice had serum FGF23 <4% of Hyp (p < 0.01), and this targeted restriction normalized serum phosphorus and ricketic bone disease. In summary, deleting FGF23 within early osteoblasts and osteocytes demonstrated that both cell types contribute to baseline circulating FGF23 concentrations, and that targeting osteoblasts/osteocytes for FGF23 production can modify systemic responses to changes in serum phosphate concentrations and rescue the Hyp genetic syndrome. © 2016 American Society for Bone and Mineral Research.
涉及Fgf23的转基因和基因敲除(KO)动物在定义矿物质代谢的新方面提供了大量信息,但受到寿命缩短、无法进行空间/时间FGF23控制以及全球KO动物不育的限制。为了更精细地测试全身和遗传影响在FGF23产生中的作用,开发了一种携带floxed(“f”)-Fgf23等位基因(外显子2被floxed)的小鼠,当与全球Cre转基因小鼠(eIIa-cre)杂交时,该等位基因在体内发生重组。重组等位基因(“Δ”)的纯合小鼠血清中未检测到完整的FGF23,血清磷酸盐升高(p < 0.05),肾脏Cyp27b1 mRNA增加(p < 0.05),这与全球Fgf23-KO小鼠相似。为了在磷酸盐刺激期间分离细胞FGF23反应,将Fgf23(Δ/f)小鼠与早期成骨细胞Iα1胶原蛋白2.3-kb启动子-Cre小鼠(Col2.3-cre)和晚期成骨细胞/早期骨细胞牙本质基质蛋白-1-Cre(Dmp1-cre)交配。与对照组相比,Fgf23(Δ/f) /Col2.3-cre(+)和Fgf23(Δ/f) /Dmp1-cre(+)小鼠的基线血清完整FGF23降低。用高磷饮食刺激后,Cre(-)小鼠的血清FGF23增加了2.1倍至2.5倍(p < 0.01),但Col2.3-cre(+)小鼠没有显著增加,尽管两种模型中都存在普遍的高磷血症,Dmp1-cre(+)小鼠的血清FGF23仅增加了37%(p < 0.01)。将Fgf23(Δ/f) /Col2.3-cre培育到Hyp(小鼠X连锁低磷血症[XLH]模型)遗传背景上,以测试成骨细胞和骨细胞对FGF23升高和Hyp疾病表型的贡献。鉴于其明显的低磷血症,Hyp小鼠的FGF23维持在不适当的高水平,而Hyp/Fgf23(Δ/f) /Col2.3-cre(+)小鼠的血清FGF23 < Hyp小鼠的4%(p < 0.01),这种靶向限制使血清磷和佝偻病骨病正常化。总之,在早期成骨细胞和骨细胞中删除FGF23表明这两种细胞类型都对基线循环FGF23浓度有贡献,并且针对成骨细胞/骨细胞产生FGF23可以改变对血清磷酸盐浓度变化的全身反应,并挽救Hyp遗传综合征。© 2016美国骨与矿物质研究协会。