Investigation of the impact of bromodomain inhibition on cytoskeleton stability and contraction.
Investigation of the impact of bromodomain inhibition on cytoskeleton stability and contraction.
复制标题
研究溴结构域抑制对细胞骨架稳定性和收缩的影响。
DOI：
10.1101/2023.11.14.567076
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Adam,RosalynM
中科院分区：
文献类型：
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作者：
Bigger-Allen,Alexander;Gheinani,AliHashemi;Adam,RosalynM
背景：心脏、血管、膀胱和肠道等收缩器官的损伤可刺激病理反应，导致正常收缩能力的丧失。血小板衍生生长因子和转化生长因子β是研究最多的损伤反应的启动者之一，已被证明可以诱导机械活动的中空器官细胞的异常收缩，包括平滑肌细胞和成纤维细胞。然而，导致血管内皮生长因子和转化生长因子β下游收缩改变的机制还不完全清楚，限制了治疗干预。方法为了确定潜在的分子靶点，我们利用公开数据的分析，比较了转化生长因子和转化生长因子β刺激的机械活性细胞的转录变化。对公开可用的数据集进行了额外的分析，研究对象是在存在或不存在MYC抑制剂JQ1的情况下处理的SMC和成纤维细胞。通过定量聚合酶链式反应、免疫印迹和胶原凝胶收缩试验对电子显微镜下的结果进行验证，以测量JQ1对机械激活细胞中细胞骨架相关基因、蛋白质和收缩能力的影响。用似然比检验和FDR校正p值确定显著差异表达基因。结果比较血小板衍生生长因子和转化生长因子β刺激的血管内皮细胞和成纤维细胞，鉴定出受MYC和AP-1转录因子复合体成员共同调控的分子图谱。此外，计算机分析揭示了一组独特的细胞骨架相关基因，这些基因对JQ1抑制MYC很敏感。结论MYC是导致成纤维细胞和成纤维细胞细胞骨架和收缩异常的重要因素，提示JQ1可用于恢复中空器官的正常收缩功能。结论JQ1可抑制β和PDGF1诱导的细胞骨架改变和细胞收缩。
BackgroundInjury to contractile organs such as the heart, vasculature, urinary bladder and gut can stimulate a pathological response that results in loss of normal contractility. PDGF and TGFβ are among the most well studied initiators of the injury response and have been shown to induce aberrant contraction in mechanically active cells of hollow organs including smooth muscle cells (SMC) and fibroblasts. However, the mechanisms driving contractile alterations downstream of PDGF and TGFβ in SMC and fibroblasts are incompletely understood, limiting therapeutic interventions.MethodsTo identify potential molecular targets, we have leveraged the analysis of publicly available data, comparing transcriptomic changes in mechanically active cells stimulated with PDGF and TGFβ. Additional Analysis of publicly available data sets were performed on SMC and fibroblasts treated in the presence or absence of the MYC inhibitor JQ1. Validation of in silico findings were performed with qPCR, immunoblots, and collagen gel contraction assays measure the effect of JQ1 on cytoskeleton associated genes, proteins and contractility in mechanically active cells. Likelihood ratio test and FDR adjustedp-values were used to determine significant differentially expressed genes. Student ttest were used to calculate statistical significance of qPCR and contractility analyses.ResultsComparing PDGF and TGFβ stimulated SMC and fibroblasts identified a shared molecular profile regulated by MYC and members of the AP-1 transcription factor complex. Additional in silico analysis revealed a unique set of cytoskeleton-associated genes that were sensitive to MYC inhibition with JQ1. In vitro validation demonstrated JQ1 was also able to attenuate TGFβ and PDGF induced changes to the cytoskeleton and contraction of smooth muscle cells and fibroblasts in vitro.ConclusionsThese findings identify MYC as a key driver of aberrant cytoskeletal and contractile changes in fibroblasts and SMC, and suggest that JQ1 could be used to restore normal contractile function in hollow organs.