FGF receptor inhibitor BGJ398 partially rescues osteoarthritis-like phenotype in older high molecular weight FGF2 transgenic mice via multiple mechanisms.
FGF receptor inhibitor BGJ398 partially rescues osteoarthritis-like phenotype in older high molecular weight FGF2 transgenic mice via multiple mechanisms.
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DOI：
10.1038/s41598-022-20269-6
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发表时间：
2022-09-24
影响因子：
4.6
通讯作者：
Xiao, Liping
中科院分区：
文献类型：
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作者：
Hurley, Marja M;Coffin, J Douglas;Doetschman, Thomas;Valera, Christina;Clarke, Kai;Xiao, Liping
我们使用碱性成纤维细胞生长因子（FGF2）转基因小鼠作为人类x连锁低磷血症（XLH）相关退行性骨关节炎（OA）的实验模型，以研究该疾病的发病机制并测试潜在的药物治疗方法。本研究检测了小分子成纤维细胞生长因子受体酪氨酸激酶（FGFRTK）抑制剂BJG398对高分子量成纤维细胞生长因子2转基因（HMWTgFGF2）小鼠膝关节骨性关节炎表型的修复作用。BJG398给8月龄雌性HMWTgFGF2小鼠体内注射6周。采用组织形态学、免疫组化和显微ct对bgj398治疗组和对照组的膝关节进行观察。我们评估：成纤维细胞生长因子23 （FGF23）表达和FGFR1活性；基质金属蛋白酶13 （MMP13）和聚合酶2 （ADAMTS5）的表达；然后通过SMAD1/5/8-pSMAD6、pERK1/2和runt相关转录因子2 （RUNX2）进行信号传导。利用PrimePCR阵列，我们确定了由BGJ398调控的TGFB/BMP2信号通路中主要靶基因的作用。BGJ398抑制HMWFGF2/ fgf23诱导的骨形态发生蛋白受体-1、骨形态发生蛋白-2和4以及丝氨酸肽酶抑制剂（分支E，成员1）的升高。显微ct和组织学结果显示，BGJ398治疗可恢复HMWTgFGF2小鼠软骨下骨和膝关节软骨的OA变化。基因表达和信号转导结果提供了令人信服的证据，HMWFGF2通过FGFRTK产生OA，并具有定义OA的下游信号，即通过BMP-BMPR增加FGF23-FGFR1活性，激活pSMAD1/5/8-RUNX2和pERK信号通路，然后上调MMP13和ADAMTS5降解基质。BGJ398处理有效地逆转了这些OA分子表型，进一步证明了HMWFGF2在转基因小鼠中产生的OA是fgfr介导的，并且表型上复制了Hyp小鼠XLH同源物中发现的OA，其中Phex （X染色体上的磷酸盐调节内肽酶）基因和人类XLH-OA的自发突变。总之，本研究的结果解释了FGF2的多效性作用是如何来自于HMW蛋白同种异构体对软骨和骨骼稳态的不同功能，以及xlh -退行性骨关节病的发病机制。BGJ398通过多种机制抑制hmwfgf2诱导的骨关节炎。这些结果为BGJ398作为XLH骨性关节炎治疗剂的潜在应用提供了重要的科学依据。
We have used Basic Fibroblast Growth Factor (FGF2) transgenic mice as experimental models for human X-linked hypophosphatemia (XLH)-related degenerative osteoarthritis (OA) to investigate the pathogenesis of the disease and to test potential pharmacotherapies for treatment. This study tested the efficacy of BJG398, a small molecule fibroblast growth factor receptor tyrosine kinase (FGFRTK) inhibitor, to rescue the knee joint osteoarthritis phenotype in High Molecular Weight fibroblast growth factor 2 transgenic (HMWTgFGF2) mice. BJG398 was administered in vivo to 8-month-old female HMWTgFGF2 mice for six weeks. Histomorphometry, immunohistochemistry and micro-CT were used to examine the knee joints in BGJ398-treated and control mice. We assessed: Fibroblast Growth Factor 23 (FGF23) expression and FGFR1 activity; Matrix metalloproteinase 13 (MMP13) and Aggrecanase2 (ADAMTS5) expression; then signaling by SMAD1/5/8-pSMAD6, pERK1/2 and Runt-related transcription factor 2 (RUNX2). Using PrimePCR arrays, we identified a contributing role for major target genes in the TGFB/BMP2 signaling pathway that were regulated by BGJ398. BGJ398 inhibited HMWFGF2/FGF23-induced increase in bone morphogenic protein receptor-1, bone morphogenic protein-2 and 4 and Serine peptidase inhibitor, clade E, member 1. The results from Micro-CT and histology show BGJ398 treatment rescued the OA changes in subchondral bone and knee articular cartilage of HMWTgFGF2 mice. The gene expression and signal transduction results provide convincing evidence that HMWFGF2 generates OA through FGFRTK with characteristic downstream signaling that defines OA, namely: increased FGF23-FGFR1 activity with BMP-BMPR, activation of pSMAD1/5/8-RUNX2 and pERK signaling pathways, then upregulation of MMP13 and ADAMTS5 to degrade matrix. BGJ398 treatment effectively reversed these OA molecular phenotypes, providing further evidence that the OA generated by HMWFGF2 in the transgenic mice is FGFR-mediated and phenocopies the OA found in the Hyp mouse homolog of XLH with a spontaneous mutation in the Phex (phosphate regulating endopeptidase on the X chromosome) gene and human XLH-OA. Overall, the results obtained here explain how the pleotropic effects of FGF2 emanate from the different functions of HMW protein isoforms for cartilage and bone homeostasis, and the pathogenesis of XLH-degenerative osteoarthropathy. BGJ398 inhibits HMWFGF2-induced osteoarthritis via multiple mechanisms. These results provided important scientific evidence for the potential application of BGJ398 as a therapeutic agent for osteoarthritis in XLH.