PDF(Pubmed)
Abstract：
UNASSIGNED: Mutations in Slc26a2 cause a spectrum of autosomal-recessive chondrodysplasia with a significant and negligible influence on the quality of life. It has been reported that Slc26a2 deficiency triggers the ATF6 branch of the UPR, which may, in turn, activate the negative regulator of the FGFR3 signaling pathway. However, the correlation between the deletion of Slc26a2 and the augmentation of downstream phosphorylation of FGFR3 has not been investigated in vivo.
UNASSIGNED: First, we constructed Slc26a2 and Fgfr3 double knockout mouse lines and observed gross views of the born mice and histological staining of the tibial growth plates. The second approach was to construct tamoxifen-inducible Cre-ERT2 mouse models to replicate SLC26A2-related non-lethal dysplastic conditions. Pharmacological intervention was performed by administering the FGFR3 inhibitor NVP-BGJ398. The effect of NVP-BGJ398 on chondrocytes was assessed by Alcian blue staining, proliferation, apoptosis, and chondrocyte-specific markers and then verified by western blotting for variations in the downstream markers of FGFR3. The growth process was detected using X-rays, micro-CT examination, histomorphometry staining of growth plates, and immunofluorescence.
UNASSIGNED: Genetic ablation of Fgfr3 in embryonic Slc26a2-deficient chondrocytes slightly attenuated chondrodysplasia. Subsequently, in the constructed mild dysplasia model, we found that postnatal intervention with Fgfr3 gene in Slc26a2-deficient chondrocytes partially alleviated chondrodysplasia. In chondrocyte assays, NVP-BGJ398 suppressed the defective phenotype of Slc26a2-deficient chondrocytes and restored the phosphorylation downstream of FGFR3 in a concentration-dependent manner. In addition, in vivo experiments showed significant alleviation of impaired chondrocyte differentiation, and micro-CT analysis showed a clear improvement in trabecular bone microarchitectural parameters.
UNASSIGNED: Our results suggested that inhibition of FGFR3 signaling pathway overactivation and NVP-BGJ398 has promising therapeutic implications for the development of SLC26A2-related skeletal diseases in humans.
UNASSIGNED: Our data provide genetic and pharmacological evidence that targeting FGFR3 signaling via NVP-BGJ398 could be a route for the treatment of SLC26A2-associated skeletal disorders, which promisingly advances translational applications and therapeutic development.
UNASSIGNED: First, we constructed Slc26a2 and Fgfr3 double knockout mouse lines and observed gross views of the born mice and histological staining of the tibial growth plates. The second approach was to construct tamoxifen-inducible Cre-ERT2 mouse models to replicate SLC26A2-related non-lethal dysplastic conditions. Pharmacological intervention was performed by administering the FGFR3 inhibitor NVP-BGJ398. The effect of NVP-BGJ398 on chondrocytes was assessed by Alcian blue staining, proliferation, apoptosis, and chondrocyte-specific markers and then verified by western blotting for variations in the downstream markers of FGFR3. The growth process was detected using X-rays, micro-CT examination, histomorphometry staining of growth plates, and immunofluorescence.
UNASSIGNED: Genetic ablation of Fgfr3 in embryonic Slc26a2-deficient chondrocytes slightly attenuated chondrodysplasia. Subsequently, in the constructed mild dysplasia model, we found that postnatal intervention with Fgfr3 gene in Slc26a2-deficient chondrocytes partially alleviated chondrodysplasia. In chondrocyte assays, NVP-BGJ398 suppressed the defective phenotype of Slc26a2-deficient chondrocytes and restored the phosphorylation downstream of FGFR3 in a concentration-dependent manner. In addition, in vivo experiments showed significant alleviation of impaired chondrocyte differentiation, and micro-CT analysis showed a clear improvement in trabecular bone microarchitectural parameters.
UNASSIGNED: Our results suggested that inhibition of FGFR3 signaling pathway overactivation and NVP-BGJ398 has promising therapeutic implications for the development of SLC26A2-related skeletal diseases in humans.
UNASSIGNED: Our data provide genetic and pharmacological evidence that targeting FGFR3 signaling via NVP-BGJ398 could be a route for the treatment of SLC26A2-associated skeletal disorders, which promisingly advances translational applications and therapeutic development.
摘要：
■Slc26a2中的突变导致一系列常染色体隐性遗传软骨发育不良，对生活质量的影响显着并且可以忽略不计。据报道，Slc26a2缺陷引发了普遍定期审议的ATF6分支，可能，反过来,激活FGFR3信号通路的负调节因子。然而,Slc26a2缺失与FGFR3下游磷酸化增强之间的相关性尚未在体内进行研究。
■首先，我们构建了Slc26a2和Fgfr3双敲除小鼠系，并观察了出生小鼠的总体视图和胫骨生长板的组织学染色。第二种方法是构建他莫昔芬诱导的Cre-ERT2小鼠模型以复制SLC26A2相关的非致死性发育不良状况。通过施用FGFR3抑制剂NVP-BGJ398进行药理学干预。通过Alcian蓝染色评估NVP-BGJ398对软骨细胞的影响，扩散,凋亡，和软骨细胞特异性标志物，然后通过蛋白质印迹验证FGFR3下游标志物的变化。使用X射线检测生长过程，Micro-CT检查,生长板的组织形态计量学染色，和免疫荧光。
■Fgfr3在胚胎Slc26a2缺陷软骨细胞中的遗传消融略微减轻了软骨发育不良。随后，在构建的轻度发育不良模型中，我们发现出生后对Slc26a2缺陷软骨细胞进行Fgfr3基因干预部分缓解了软骨发育不良.在软骨细胞检测中，NVP-BGJ398抑制Slc26a2缺陷软骨细胞的缺陷表型，并以浓度依赖性方式恢复FGFR3下游的磷酸化。此外,体内实验显示软骨细胞分化受损的显著缓解，和micro-CT分析显示骨小梁微结构参数明显改善。
■我们的结果表明，抑制FGFR3信号通路过度激活和NVP-BGJ398对人类SLC26A2相关骨骼疾病的发展具有有希望的治疗意义。
■我们的数据提供了遗传和药理学证据，表明通过NVP-BGJ398靶向FGFR3信号可能是治疗SLC26A2相关骨骼疾病的途径，有望推进转化应用和治疗开发。
■首先，我们构建了Slc26a2和Fgfr3双敲除小鼠系，并观察了出生小鼠的总体视图和胫骨生长板的组织学染色。第二种方法是构建他莫昔芬诱导的Cre-ERT2小鼠模型以复制SLC26A2相关的非致死性发育不良状况。通过施用FGFR3抑制剂NVP-BGJ398进行药理学干预。通过Alcian蓝染色评估NVP-BGJ398对软骨细胞的影响，扩散,凋亡，和软骨细胞特异性标志物，然后通过蛋白质印迹验证FGFR3下游标志物的变化。使用X射线检测生长过程，Micro-CT检查,生长板的组织形态计量学染色，和免疫荧光。
■Fgfr3在胚胎Slc26a2缺陷软骨细胞中的遗传消融略微减轻了软骨发育不良。随后，在构建的轻度发育不良模型中，我们发现出生后对Slc26a2缺陷软骨细胞进行Fgfr3基因干预部分缓解了软骨发育不良.在软骨细胞检测中，NVP-BGJ398抑制Slc26a2缺陷软骨细胞的缺陷表型，并以浓度依赖性方式恢复FGFR3下游的磷酸化。此外,体内实验显示软骨细胞分化受损的显著缓解，和micro-CT分析显示骨小梁微结构参数明显改善。
■我们的结果表明，抑制FGFR3信号通路过度激活和NVP-BGJ398对人类SLC26A2相关骨骼疾病的发展具有有希望的治疗意义。
■我们的数据提供了遗传和药理学证据，表明通过NVP-BGJ398靶向FGFR3信号可能是治疗SLC26A2相关骨骼疾病的途径，有望推进转化应用和治疗开发。
