Steroid Resistance in Nephrotic Syndrome

肾病综合征的类固醇抵抗

• 批准号: 6967183
• 负责人: DIEGO H AVILES
• 金额: $ 12.6万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-08 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6967183
• 关键词: T lymphocyte; apoptosis; cell sorting; clinical research; corticosteroid receptors; dexamethasone; enzyme linked immunosorbent assay; flow cytometry; gel mobility shift assay; glucocorticoids; hormone sensitivity /resistance; human subject; immunopathology; immunoprecipitation; immunosuppression; interleukin 2; nephrotic syndrome; nuclear factor kappa beta; nuclear transport; polymerase chain reaction; tissue /cell culture; transcription factor

DESCRIPTION: Steroid-resistant idiopathic nephrotic syndrome (SRINS) is an important cause of renal failure and end-stage renal disease in children. The cause of steroid resistance is unknown but evidence suggests that T cell dysfunction is responsible. Based on preliminary data on abnormal T cell NF-kB isoform composition in SRINS, the proposed project will identify the defects in T cell signaling mechanisms that lead to glucocorticoid resistance in patients with SRINS. The experiments will compare T cells from newly diagnosed SRINS patients with those from steroid-sensitive patients and healthy control children. Potential defects in the following specific signal transduction mechanisms, which may be associated with abnormal NF-kB function, will be evaluated: 1) Nuclear translocation of the glucocorticoid receptor (GCR) in response to dexamethasone exposure in vitro; 2) Increased production of STAT5 and interference of nuclear transfer of GCR by heterodimerizing with STAT5; 3) Dexamethasone-stimulated IL-2 production; 4) Induction of apoptosis in response to dexamethasone exposure. The methods will include standard T cell isolation, real time RT-PCR for gene expression, electrophoretic mobility shift assay for quantifying factors in nuclear vs cytoplasmic fractions, immunoprecipitation assay to detect complexed GCRs, as well as apoptosis

描述： 类固醇抵抗性特发性肾病综合征（SRINS）是儿童肾功能衰竭和终末期肾病的重要原因。类固醇抵抗的原因尚不清楚，但有证据表明 T 细胞功能障碍是造成这一现象的原因。基于 SRINS 中异常 T 细胞 NF-kB 亚型组成的初步数据，拟议项目将确定导致 SRINS 患者糖皮质激素抵抗的 T 细胞信号传导机制缺陷。实验将比较新诊断的 SRINS 患者的 T 细胞与类固醇敏感患者和健康对照儿童的 T 细胞。将评估以下可能与 NF-kB 功能异常相关的特定信号转导机制的潜在缺陷： 1) 糖皮质激素受体 (GCR) 响应体外地塞米松暴露而发生核易位； 2)通过与STAT5异二聚化增加STAT5的产量并干扰GCR的核转移； 3）地塞米松刺激IL-2的产生； 4）响应地塞米松暴露诱导细胞凋亡。这些方法将包括标准 T 细胞分离、用于基因表达的实时 RT-PCR、用于量化核与细胞质部分中因子的电泳迁移率变化测定、用于检测复合 GCR 的免疫沉淀测定以及细胞凋亡