MOLECULAR BASIS AND MECHANISMS OF CYST ACTIVATING FACTOR

摘要多囊肾病(PKD)是导致终末期肾病的主要原因。 疾病(ESRD)。在最常见的遗传形式中，常染色体显性遗传性PKD， 囊是由编码蛋白质(多囊蛋白)的突变基因引起的。 刺激上皮细胞增殖、液体堆积和 细胞外基质重塑。尽管所有多囊肾都增大 由于囊腔内不断扩大，只有大约50%的患者会发展成 ESRD。有证据表明，表观遗传因素在 确定ADPKD的病程。在系统地寻找潜在的 “进展因子”，一种刺激上皮细胞的强效中性脂质 细胞增殖、跨上皮液分泌和单核细胞 最近在ADPKD患者的囊液中发现了趋化作用。 鉴于这些特征，这种类脂或类脂家族被命名为 包囊激活因子(CAF)。这个项目将检验这一假设 CAP是ADPKD和ADPKD肾囊肿形成的一种有效的脂质产物 作为一种进展因子，通过以下方式加速发病 促进细胞增殖，刺激上皮液 分泌，促进间质炎症和纤维化。 本方案的目的是：1)从ADPKD中提纯到均一的CaF 囊液和确定其分子结构的适当方法 分析、生化和生物物理技术；2)探索 CAF促进ADPKD囊内液体转运的机制 上皮细胞；3)阐明CAF刺激包囊的机制。 4)研究CAF对血管内皮细胞增殖的影响。 ADPKD囊上皮细胞与单核/巨噬细胞的相互作用 以及基质金属蛋白酶和金属蛋白酶抑制剂的作用 在这种互动中。 这些目标的实现需要将技术应用于 分析化学、细胞生物学、细胞生理学、分子生物学和 生物化学。预计研究结果将产生新的见解。 关于一种新发现的对人类健康具有特殊重要性的肾脏脂质 多囊肾病的发病机制。

Polycystic kidney disease (PKD) is a major cause of end-stage renal disease (ESRD). In the most common inherited form, autosomal dominant PKD, cysts are initiated by mutated genes that encode proteins (polycystins) that stimulate epithelial proliferation, fluid accumulation and extracellular matrix remodelling. Although all polycystic kidneys enlarge due to expanding cysts within them, only about 50% of patients develop ESRD. Evidence indicates that epigenetic factors have significant roles in determining the course of ADPKD. In a systematic search for potential "progression factors", a potent neutral lipid that stimulates epithelial cell proliferation, transepithelial fluid secretion and monocyte chemotaxis was recently identified in cyst fluids of patients with ADPKD. In view of these features this lipid or family of lipids has been named cyst activating factor (CAF). This project will test the hypothesis that CAP is a potent lipid product of renal cyst formation in ADPKD and functions as a progression factor that accelerates pathogenesis by increasing cellular proliferation, stimulating transpithelial fluid secretion, and promoting interstitial inflammation and fibrosis. The aims of this proposal are to: 1) Purify to homogeneity CAF from ADPKD cyst fluid and determine its molecular structure by appropriate analytical, biochemical, and biophysical techniques; 2) Explore the mechanisms by which CAF stimulates fluid transport in ADPKD cyst epithelial cells; 3) Delineate the mechanisms by which CAF stimulates cyst epithelial cell proliferation; 4) Investigate the effect of CAF on the interactions between ADPKD cyst epithelial cells and monocyte/macrophages and the role of matrix metalloproteinases and metalloproteinase inhibitors in this interaction. The achievement of these aims requires the application of techniques in analytical chemistry, cell biology, cell physiology, molecular biology and biochemistry. It is anticipated that the results will yield novel insights about a newly recognized renal lipid of specific importance to the pathogenesis of polycystic kidney disease.