Department of Pharmacology, Brain Korea 21 PLUS Project for Medical Sciences, Yonsei University College of Medicine, Seoul, 03722, Korea.
Department of Medicine, Division of Nephrology, Boston Children's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
Exp Mol Med. 2020 Apr;52(4):594-603. doi: 10.1038/s12276-020-0410-4. Epub 2020 Apr 1.
PLCE1 encodes phospholipase C epsilon, and its mutations cause recessive nephrotic syndrome. However, the mechanisms by which PLCE1 mutations result in defects associated with glomerular function are not clear. To address this, we investigated the function of PLCE1 in podocytes called glomerular epithelial cells, where the pathogenesis of nephrotic syndrome converges. PLCE1 colocalized with Rho GTPases in glomeruli. Further, it interacted with Rho GTPases through the pleckstrin homology domain and Ras GTP-binding domains 1/2. Knockdown or knockout of PLCE1 in podocytes resulted in decreased levels of GTP-bound Rac1 and Cdc42, but not those of RhoA, and caused a reduction in cell migration. PLCE1 interacted with NCK2 but not with NCK1. Similar to the PLCE1 knockout, NCK2 knockout resulted in decreased podocyte migration. Knockout of PLCE1 reduced the EGF-induced activation of ERK and cell proliferation in podocytes, whereas knockout of NCK2 did not affect proliferation. Further, the knockout of PLCE1 also resulted in decreased expression of podocyte markers, including NEPH1, NPHS1, WT1, and SYNPO, upon differentiation, but the knockout of NCK2 did not affect the expression of these markers. Therefore, our findings demonstrate that PLCE1 regulates Rho GTPase activity and cell migration through interacting with NCK2 and that PLCE1 also plays a role in the proliferation and differentiation of podocytes, regardless of the presence of NCK2.
PLCE1 编码磷脂酶 C ɛ,其突变导致常染色体隐性遗传性肾病综合征。然而,PLCE1 突变导致与肾小球功能相关缺陷的机制尚不清楚。为了解决这个问题,我们研究了 PLCE1 在足细胞(也称为肾小球上皮细胞)中的功能,肾病综合征的发病机制在此汇聚。PLCE1 在肾小球中与 Rho GTPases 共定位。此外,它通过 PH 结构域和 Ras GTP 结合结构域 1/2 与 Rho GTPases 相互作用。在足细胞中敲低或敲除 PLCE1 会导致 GTP 结合的 Rac1 和 Cdc42 水平降低,但 RhoA 水平不变,并且细胞迁移减少。PLCE1 与 NCK2 相互作用而不与 NCK1 相互作用。与 PLCE1 敲除相似,NCK2 敲除也导致足细胞迁移减少。PLCE1 敲除减少了 EGF 诱导的足细胞 ERK 激活和细胞增殖,而 NCK2 敲除则不影响增殖。此外,PLCE1 敲除还导致足细胞标志物的表达减少,包括分化时的 NEPH1、NPHS1、WT1 和 SYNPO,但 NCK2 敲除不影响这些标志物的表达。因此,我们的研究结果表明,PLCE1 通过与 NCK2 相互作用调节 Rho GTPase 活性和细胞迁移,并且 PLCE1 还在足细胞的增殖和分化中发挥作用,而与 NCK2 的存在与否无关。