Determining the Role of Canonical Notch Signalling in the Regulation of the Glucocorticoid Receptor in Steroid Resistant Nephrotic Syndrome

确定经典Notch信号传导在类固醇抵抗性肾病综合征糖皮质激素受体调节中的作用

• 批准号: MR/K010654/1
• 负责人: Aoife Waters
• 金额: $ 115.06万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FK010654%2F1
• 关键词: Determining; Role; Canonical; Notch; Signalling

Scarring of the glomerulus can cause terminal kidney disease. Glucocorticoid treatment has a beneficial effect but resistance (GR) develops over time. Alternative treatments are available with significant side effects and only 30-60% of patients respond. Consequently, new treatments are needed. Recently, we have shown that activation of a critical signalling pathway (Notch) is involved in initiating glomerular scarring. Using knockout mouse models and drugs which block the pathway, we and others, have shown that glomerular scarring can be prevented. Studies in leukaemic patients have shown that Notch activity can inhibit the glucocorticoid response, an effect which can be reversed by inhibiting Notch. My preliminary data shows that the glucocorticoid receptor, NR3C1, is expressed in control human glomerular cells in vitro and in vivo and is downregulated in human glomerular cells derived from patients with steroid-resistance. As activation of the Notch effector gene, HES-1, has been shown to downregulate NR3C1 and mediate the development of GR in leukaemia, I have observed that HES-1 is temporally upregulated in podocytes of sclerosed glomeruli in kidney biopsy tissue in patients with nephrotic syndrome who have been initially glucocorticoid sensitive and subsequently developed GR. These data support a role for Notch activation in steroid-resistant nephrotic syndrome.My research objectives will involve testing a few basic principles.Firstly, I would like to demonstrate that glucocorticoid-responsive gene expression occurs in glomerular epithelial cells when treated with glucocorticoid therapy and then determine, absence of expression of the same genes in glomerular cells derived from kidney biopsies of steroid resistant nephrotic syndrome (SRNS) patients. Next, I propose to determine whether expression of components of the Notch signalling pathway are upregulated in podocyte cell lines of SRNS patients. If this is the case, I will then deplete Notch effector genes to determine if I can rescue expression of glucocorticoid-responsive genes.Secondly, I will use mouse models of genetic forms of glomerulosclerosis (scarring of the glomerulus), to determine whether loss of glucocorticoid-responsive gene expression in glomerular epithelial cells is associated with the development of glomerulosclerosis and whether this is also associated with an up regulation of Notch pathway components.I will next specifically explore whether secondary Notch activation in mouse models of genetic forms of glomerulosclerosis is associated with loss of glucocorticoid-responsive gene expression in glomerular epithelial cells - this will be facilitated by generating transgenic mice of Notch activation and examining whether onset of glomerular scarring as a result of Notch activation is associated with loss of glucocorticoid-responsive gene expression in glomerular epithelial cells. If this proves true, chemicals which inhibit the Notch pathway will be used in mouse models of genetic forms of glomerulosclerosis to see whether it is possible to rescue glucocorticoid-responsiveness concommitant with resolution of glomerular scarring.In parallel, genetic analysis of a large cohort of patients with nephrotic syndrome will be undertaken with collaborators in an attempt to further define molecular mechanisms of glomerulosclerosis which may open up new avenues of investigation for therapeutic strategies in nephrotic syndrome.

肾小球瘢痕化可导致终末期肾病。糖皮质激素治疗具有有益效果，但随着时间的推移会产生耐药性（GR）。替代治疗具有显著的副作用，只有30-60%的患者有反应。因此，需要新的治疗方法。最近，我们已经表明，激活的关键信号通路（Notch）参与启动肾小球瘢痕形成。利用基因敲除小鼠模型和阻断该途径的药物，我们和其他人已经证明肾小球瘢痕形成是可以预防的。在白血病患者中的研究表明，Notch活性可以抑制糖皮质激素反应，这种作用可以通过抑制Notch逆转。我的初步数据显示，糖皮质激素受体，NR 3C 1，在体外和体内的控制人类肾小球细胞中表达，并下调来自类固醇耐药患者的人类肾小球细胞。由于Notch效应基因HES-1的激活已显示下调NR 3C 1并介导白血病中GR的发展，我已经观察到，在肾病综合征患者的肾活检组织中，HES-1在硬化肾小球的足细胞中暂时上调，这些患者最初是糖皮质激素敏感的，随后发展为GR。耐药肾病综合征。我的研究目标将涉及测试一些基本原则。首先，我想证明糖皮质激素反应基因表达发生在肾小球上皮细胞时，用糖皮质激素治疗，然后确定，在来源于类固醇耐药肾病综合征（SRNS）患者的肾活检的肾小球细胞中缺乏相同基因的表达。接下来，我建议确定是否表达的Notch信号通路的组件在SRNS患者的足细胞系上调。如果是这样的话，我将耗尽Notch效应基因，以确定我是否可以挽救糖皮质激素反应基因的表达。其次，我将使用肾小球硬化症遗传形式的小鼠模型（肾小球瘢痕形成），以确定是否糖皮质激素的损失-肾小球上皮细胞中的反应性基因表达与肾小球硬化的发展有关，以及这是否也与肾小球硬化的上调有关。Notch途径组分。接下来，我将具体探索肾小球硬化症遗传形式的小鼠模型中的继发性Notch激活是否与肾小球上皮细胞中糖皮质激素应答基因表达的丧失相关-这将通过产生Notch激活的转基因小鼠并检查Notch激活导致的肾小球瘢痕形成的发作是否与糖皮质激素的丧失相关来促进。肾小球上皮细胞中的应答基因表达。如果这被证明是真的，抑制Notch途径的化学物质将被用于肾小球硬化症遗传形式的小鼠模型中，以观察是否有可能在解决肾小球瘢痕形成的同时拯救糖皮质激素反应性。将与合作者一起对一大批肾病综合征患者进行遗传分析，试图进一步确定肾小球硬化的分子机制，肾病综合征治疗策略研究的新途径。

项目成果

The clinical spectrum of hemolytic uremic syndrome secondary to complement factor H autoantibodies.

补体因子 H 自身抗体继发的溶血性尿毒症综合征的临床谱。

• DOI: 10.5414/cn107777
• 发表时间: 2015
• 期刊: Clinical nephrology
• 影响因子: 1.1
• 作者: Kim JJ

Clinico-pathological correlations of congenital and infantile nephrotic syndrome over twenty years.

• DOI: 10.1007/s00467-014-2856-x
• 发表时间: 2014-11
• 期刊: PEDIATRIC NEPHROLOGY
• 影响因子: 3
• 作者: Kari, Jameela A.;Montini, Giovanni;Bockenhauer, Detlef;Brennan, Eileen;Rees, Lesley;Trompeter, Richard S.;Tullus, Kjell;van't Hoff, William;Waters, Aoife;Ashton, Emma;Lench, Nicholas;Sebire, Neil J.;Marks, Stephen D.
• 通讯作者: Marks, Stephen D.

Malformations of the Murine Kidney are associated with loss of Cenp-F function

小鼠肾脏畸形与 Cenp-F 功能丧失有关

• 发表时间: 2018
• 作者: Chanell O Haley

Outcome of adult patients with X-linked hypophosphatemia caused by PHEX gene mutations.

• DOI: 10.1007/s10545-018-0147-6
• 发表时间: 2018-09
• 期刊: Journal of inherited metabolic disease
• 影响因子: 4.2
• 作者: Chesher D;Oddy M;Darbar U;Sayal P;Casey A;Ryan A;Sechi A;Simister C;Waters A;Wedatilake Y;Lachmann RH;Murphy E
• 通讯作者: Murphy E

Activation of podocyte Notch mediates early Wt1 glomerulopathy.

• DOI: 10.1016/j.kint.2017.11.014
• 发表时间: 2018-04
• 期刊: Kidney international
• 影响因子: 19.6
• 作者: Asfahani RI;Tahoun MM;Miller-Hodges EV;Bellerby J;Virasami AK;Sampson RD;Moulding D;Sebire NJ;Hohenstein P;Scambler PJ;Waters AM
• 通讯作者: Waters AM