clear factor kappa-B kinase subunit beta to inhibit NF-κB activation, which, subsequently restores Krüppel-like factor 15 under cell stress. Background Podocyte loss is the major driver of primary glomerular diseases such as FSGS. While systemic glucocorticoids remain the initial and primary therapy for these diseases, high-dose and chronic use of glucocorticoids is riddled with systemic toxicities. Krüppel-like factor 15 (KLF15) is a glucocorticoid-responsive gene, which is essential for the restoration of mature podocyte differentiation markers and stabilization of actin cytoskeleton in the setting of cell stress. Induction of KLF15 attenuates podocyte injury and glomerulosclerosis in the setting of cell stress. Methods A cell-based high-throughput screen with a subsequent structure–activity relationship study was conducted to identify novel agonists of KLF15 in human podocytes. Next, the agonist was tested in cultured human podocytes under cell stress and in three independent proteinuric models (LPS, nephrotoxic serum nephritis, and HIV-1 transgenic mice). A combination of RNA sequencing and molecular modeling with experimental validation was conducted to demonstrate the direct target of the agonist. Results The high-throughput screen with structure–activity relationship study identified BT503, a urea-based compound, as a novel agonist of KLF15, independent of glucocorticoid signaling. BT503 demonstrated protective effects in cultured human podocytes and in three independent proteinuric murine models. Subsequent molecular modeling with experimental validation shows that BT503 targets the inhibitor of nuclear factor kappa-B kinase complex by directly binding to inhibitor of nuclear factor kappa-B kinase subunit beta to inhibit canonical NF-κB signaling, which, in turn, restores KLF15 under cell stress, thereby rescuing podocyte loss and ameliorating kidney injury. Conclusions By developing and validating a cell-based high-throughput screen in human podocytes, we identified a novel agonist for KLF15 with salutary effects in proteinuric murine models through direct inhibition of inhibitor of nuclear factor kappa-B kinase subunit beta kinase activity....

中文翻译：

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蛋白尿性肾病中 Krüppel 样因子 15 的小分子激动剂


清除因子 kappa-B 激酶亚基 β 抑制 NF-κB 激活，随后在细胞应激下恢复 Krüppel 样因子 15。背景 足细胞丢失是 FSGS 等原发性肾小球疾病的主要驱动因素。虽然全身性糖皮质激素仍然是这些疾病的初始和主要治疗，但大剂量和长期使用糖皮质激素充满了全身毒性。Krüppel 样因子 15 （KLF15） 是一种糖皮质激素反应基因，在细胞应激情况下，它对于成熟足细胞分化标志物的恢复和肌动蛋白细胞骨架的稳定至关重要。在细胞应激的情况下，KLF15 的诱导可减轻足细胞损伤和肾小球硬化。方法 进行基于细胞的高通量筛选和随后的构效关系研究，以鉴定人足细胞中 KLF15 的新型激动剂。接下来，在细胞应激下培养的人足细胞和三种独立的蛋白尿模型 （LPS、肾毒性血清肾炎和 HIV-1 转基因小鼠） 中测试激动剂。将 RNA 测序和分子建模与实验验证相结合，以证明激动剂的直接靶标。结果 高通量筛选与构效关系研究确定 BT503（一种基于尿素的化合物）是 KLF15 的新型激动剂，独立于糖皮质激素信号传导。BT503 在培养的人足细胞和三个独立的蛋白尿小鼠模型中表现出保护作用。 随后的分子建模和实验验证表明，BT503 通过直接与核因子 kappa-B 激酶亚基 β 结合来靶向核因子 kappa-B 激酶复合物的抑制剂，以抑制经典的 NF-κB 信号传导，进而在细胞应激下恢复 KLF15，从而挽救足细胞丢失并改善肾损伤。结论 通过开发和验证基于细胞的人足细胞高通量筛选，我们通过直接抑制核因子 kappa-B 激酶亚基 β 激酶活性的抑制剂，在蛋白尿小鼠模型中确定了一种具有有益作用的新型 KLF15 激动剂。