Modulated Fibrosis and Mechanosensing of Fibroblasts by SB525334 in Pediatric Subglottic Stenosis
Modulated Fibrosis and Mechanosensing of Fibroblasts by SB525334 in Pediatric Subglottic Stenosis
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SB525334 在儿科声门下狭窄中调节成纤维细胞的纤维化和机械传感
DOI：
10.1002/lary.30873
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Jacobs, Ian N.
中科院分区：
文献类型：
--
作者：
Ali Akbari Ghavimi, Soheila;Aronson, Matthew R.;Ghaderi, Daniel D.;Friedman, Ryan M.;Patel, Neil;Giordano, Terri;Borek, Ryan C.;Devine, Conor M.;Han, Lin;Jacobs, Ian N.
声门下狭窄（SGS）可能是由于长时间插管导致的，其中纤维化瘢痕组织使气道变窄。瘢痕由分化的肌成纤维细胞分泌过量的细胞外基质（ECM）形成。TGF-β1被广泛接受为纤维化的调节因子;然而，尚不清楚生物力学途径如何共同调节纤维化。因此，我们对SGS儿科患者的成纤维细胞进行表型分析，以探索关键信号通路TGF-β和Hippo，影响瘢痕形成，并评估潜在治疗性小分子SB 525334和DRD 1激动剂盐酸二氢屈西定（DHX）抑制这些通路的影响方法：对从声门下狭窄进展和成熟患者的声门下以及远端对照活检组织中分离的喉成纤维细胞进行评估，α-平滑肌肌动蛋白免疫染色和α-SMA、FN、HGF和CTGF标志物的基因表达。在TGF-β1诱导的纤维化过程中使用抑制剂SB 525334或DHX调节TGF-β和Hippo信号通路，并通过RT-qPCR分析差异基因表达和原子力显微镜分析ECM stiffen.ResultsSGS成纤维细胞在TGF-β1刺激后表现出更高的α-SMA染色和更高的炎性细胞因子和纤维化标记物表达（p< 0.05）。SB 525334通过减少SMAD 2/3核易位（p< 0.0001）和促纤维化基因表达（p< 0.05）将水平恢复至基线。狭窄成纤维细胞的ECM硬度大于健康成纤维细胞，并且通过使用SB 525334和DHX的Hippo通路调节恢复到基线（p< 0.01）。结论我们证明了来自儿科SGS患者患病和健康区域的不同成纤维细胞表型对TGF-β1刺激的反应不同，SB 525334通过同时调节TGF-β和Hippo信号通路而具有上级的声门下狭窄治疗潜力。
ObjectiveSubglottic stenosis (SGS) may result from prolonged intubation where fibrotic scar tissue narrows the airway. The scar forms by differentiated myofibroblasts secreting excessive extracellular matrix (ECM). TGF‐β1 is widely accepted as a regulator of fibrosis; however, it is unclear how biomechanical pathways co‐regulate fibrosis. Therefore, we phenotyped fibroblasts from pediatric patients with SGS to explore how key signaling pathways, TGF‐β and Hippo, impact scarring and assess the impact of inhibiting these pathways with potential therapeutic small molecules SB525334 and DRD1 agonist dihydrexidine hydrochloride (DHX).MethodsLaryngeal fibroblasts isolated from subglottic as well as distal control biopsies of patients with evolving and maturing subglottic stenosis were assessed by α‐smooth muscle actin immunostaining and gene expression for α‐SMA, FN, HGF, and CTGF markers. TGF‐β and Hippo signaling pathways were modulated during TGF‐β1‐induced fibrosis using the inhibitor SB525334 or DHX and analyzed by RT‐qPCR for differential gene expression and atomic force microscopy for ECM stiffness.ResultsSGS fibroblasts exhibited higher α‐SMA staining and greater inflammatory cytokine and fibrotic marker expression upon TGF‐β1 stimulation (p< 0.05). SB525334 restored levels to baseline by reducing SMAD2/3 nuclear translocation (p< 0.0001) and pro‐fibrotic gene expression (p< 0.05). ECM stiffness of stenotic fibroblasts was greater than healthy fibroblasts and was restored to baseline by Hippo pathway modulation using SB525334 and DHX (p< 0.01).ConclusionWe demonstrate that distinct fibroblast phenotypes from diseased and healthy regions of pediatric SGS patients respond differently to TGF‐β1 stimulation, and SB525334 has the superior potential for subglottic stenosis treatment by simultaneously modulating TGF‐β and Hippo signaling pathways.Level of EvidenceNALaryngoscope, 134:287–296, 2024
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9.7
作者：
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