Department of Medicine, University of Texas Health Science Center, San Antonio, Texas, USA.
Geriatric Research, Education, and Clinical Center, South Texas Veterans Health Care System, San Antonio, Texas, USA.
J Biol Chem. 2020 Oct 16;295(42):14262-14278. doi: 10.1074/jbc.RA120.014994. Epub 2020 Jul 30.
Interaction of transforming growth factor-β (TGFβ)-induced canonical signaling with the noncanonical kinase cascades regulates glomerular hypertrophy and matrix protein deposition, which are early features of glomerulosclerosis. However, the specific target downstream of the TGFβ receptor involved in the noncanonical signaling is unknown. Here, we show that TGFβ increased the catalytic loop phosphorylation of platelet-derived growth factor receptor β (PDGFRβ), a receptor tyrosine kinase expressed abundantly in glomerular mesangial cells. TGFβ increased phosphorylation of the PI 3-kinase-interacting Tyr-751 residue of PDGFRβ, thus activating Akt. Inhibition of PDGFRβ using a pharmacological inhibitor and siRNAs blocked TGFβ-stimulated phosphorylation of proline-rich Akt substrate of 40 kDa (PRAS40), an intrinsic inhibitory component of mTORC1, and prevented activation of mTORC1 in the absence of any effect on Smad 2/3 phosphorylation. Expression of constitutively active myristoylated Akt reversed the siPDGFRβ-mediated inhibition of mTORC1 activity; however, co-expression of the phospho-deficient mutant of PRAS40 inhibited the effect of myristoylated Akt, suggesting a definitive role of PRAS40 phosphorylation in mTORC1 activation downstream of PDGFRβ in mesangial cells. Additionally, we demonstrate that PDGFRβ-initiated phosphorylation of PRAS40 is required for TGFβ-induced mesangial cell hypertrophy and fibronectin and collagen I (α2) production. Increased activating phosphorylation of PDGFRβ is also associated with enhanced TGFβ expression and mTORC1 activation in the kidney cortex and glomeruli of diabetic mice and rats, respectively. Thus, pursuing TGFβ noncanonical signaling, we identified how TGFβ receptor I achieves mTORC1 activation through PDGFRβ-mediated Akt/PRAS40 phosphorylation to spur mesangial cell hypertrophy and matrix protein accumulation. These findings provide support for targeting PDGFRβ in TGFβ-driven renal fibrosis.
转化生长因子-β(TGFβ)诱导的经典信号与非经典激酶级联的相互作用调节肾小球肥大和基质蛋白沉积,这是肾小球硬化的早期特征。然而,TGFβ 受体参与非经典信号的特定下游靶标尚不清楚。在这里,我们表明 TGFβ 增加了血小板衍生生长因子受体β(PDGFRβ)的催化环磷酸化,PDGFRβ 是一种在肾小球系膜细胞中大量表达的受体酪氨酸激酶。TGFβ 增加了 PDGFRβ 的 PI3-激酶相互作用的 Tyr-751 残基的磷酸化,从而激活了 Akt。使用药理学抑制剂和 siRNA 抑制 PDGFRβ 阻断了 TGFβ 刺激的富含脯氨酸的 Akt 底物 40 kDa(PRAS40)的磷酸化,PRAS40 是 mTORC1 的内在抑制成分,并且在没有对 Smad 2/3 磷酸化产生任何影响的情况下阻止了 mTORC1 的激活。组成型激活的 myristoylated Akt 的表达逆转了 siPDGFRβ 介导的 mTORC1 活性抑制;然而,磷酸缺陷型 PRAS40 的共表达抑制了 myristoylated Akt 的作用,这表明 PRAS40 磷酸化在 PDGFRβ 介导的系膜细胞中 mTORC1 激活中具有明确的作用。此外,我们证明 PDGFRβ 起始的 PRAS40 磷酸化是 TGFβ 诱导的系膜细胞肥大和纤维连接蛋白和胶原 I(α2)产生所必需的。PDGFRβ 激活磷酸化的增加也与糖尿病小鼠和大鼠肾脏皮质和肾小球中 TGFβ 表达和 mTORC1 激活的增强相关。因此,通过追踪 TGFβ 的非经典信号,我们确定了 TGFβ 受体 I 如何通过 PDGFRβ 介导的 Akt/PRAS40 磷酸化实现 mTORC1 激活,从而刺激系膜细胞肥大和基质蛋白积累。这些发现为靶向 TGFβ 驱动的肾纤维化中的 PDGFRβ 提供了支持。