School of Biological Sciences, University of Reading, Reading, U.K.
Molecular and Clinical Sciences Institute, St. George's University of London, London, U.K.
Biochem J. 2022 Feb 11;479(3):401-424. doi: 10.1042/BCJ20210615.
The extracellular signal-regulated kinase 1/2 (ERK1/2) cascade promotes cardiomyocyte hypertrophy and is cardioprotective, with the three RAF kinases forming a node for signal integration. Our aims were to determine if BRAF is relevant for human heart failure, whether BRAF promotes cardiomyocyte hypertrophy, and if Type 1 RAF inhibitors developed for cancer (that paradoxically activate ERK1/2 at low concentrations: the 'RAF paradox') may have the same effect. BRAF was up-regulated in heart samples from patients with heart failure compared with normal controls. We assessed the effects of activated BRAF in the heart using mice with tamoxifen-activated Cre for cardiomyocyte-specific knock-in of the activating V600E mutation into the endogenous gene. We used echocardiography to measure cardiac dimensions/function. Cardiomyocyte BRAFV600E induced cardiac hypertrophy within 10 d, resulting in increased ejection fraction and fractional shortening over 6 weeks. This was associated with increased cardiomyocyte size without significant fibrosis, consistent with compensated hypertrophy. The experimental Type 1 RAF inhibitor, SB590885, and/or encorafenib (a RAF inhibitor used clinically) increased ERK1/2 phosphorylation in cardiomyocytes, and promoted hypertrophy, consistent with a 'RAF paradox' effect. Both promoted cardiac hypertrophy in mouse hearts in vivo, with increased cardiomyocyte size and no overt fibrosis. In conclusion, BRAF potentially plays an important role in human failing hearts, activation of BRAF is sufficient to induce hypertrophy, and Type 1 RAF inhibitors promote hypertrophy via the 'RAF paradox'. Cardiac hypertrophy resulting from these interventions was not associated with pathological features, suggesting that Type 1 RAF inhibitors may be useful to boost cardiomyocyte function.
细胞外信号调节激酶 1/2(ERK1/2)级联反应促进心肌细胞肥大,具有心脏保护作用,三种 RAF 激酶形成信号整合的节点。我们的目的是确定 BRAF 是否与人类心力衰竭相关,BRAF 是否促进心肌细胞肥大,以及为癌症开发的 1 型 RAF 抑制剂(在低浓度下悖论性地激活 ERK1/2:“RAF 悖论”)是否可能具有相同的效果。与正常对照组相比,心力衰竭患者的心脏样本中 BRAF 上调。我们使用经他莫昔芬激活的 Cre 在心脏中评估激活的 BRAF 的作用,用于将激活的 V600E 突变的内源性基因中的心肌细胞特异性敲入。我们使用超声心动图测量心脏尺寸/功能。心肌细胞 BRAFV600E 在 10d 内诱导心脏肥大,导致射血分数和分数缩短在 6 周内增加。这与心肌细胞大小增加而没有明显纤维化一致,与代偿性肥大一致。实验性 1 型 RAF 抑制剂 SB590885 和/或 encorafenib(临床上使用的 RAF 抑制剂)增加了心肌细胞中 ERK1/2 的磷酸化,并促进了肥大,与“RAF 悖论”效应一致。两者都在体内促进了小鼠心脏的心肌肥大,心肌细胞增大而没有明显的纤维化。总之,BRAF 可能在人类衰竭心脏中发挥重要作用,激活 BRAF 足以诱导肥大,而 1 型 RAF 抑制剂通过“RAF 悖论”促进肥大。这些干预措施引起的心肌肥大与病理特征无关,这表明 1 型 RAF 抑制剂可能有助于增强心肌细胞功能。