Role of Sphingolipids in the Development of Diabetic Nephropathy

描述（由申请人提供）：鞘脂已成为参与多种细胞功能调节的关键信号分子。大多数关于鞘脂的研究都集中在神经酰胺和S1P两种生物活性鞘脂的细胞内信号功能上，但很少关注这些分子向细胞传递的来源。拟开展的研究将不仅关注这些鞘脂质的细胞外转运，还将关注血浆、尿液和分离脂蛋白中鞘磷脂和复杂的己糖基和乳糖神经酰胺分子种的细胞外转运，此外，还将确定与糖尿病和肾病相关的鞘磷脂转运变化。从633名1型糖尿病患者进入糖尿病控制和并发症试验（DCCT）时获得的血浆脂质组学分析表明，几种神经酰胺的低血浆浓度可预测大量蛋白尿的发生。相比之下，尽管糖尿病患者的鞘氨醇-1-磷酸（S1P）水平比匹配的非糖尿病受试者高2.6倍，但没有影响。1型糖尿病合并蛋白尿患者血浆载脂蛋白M (ApoM) （S1P的生理载体）浓度显著升高，且在这些患者的尿液中发现ApoM，而在正常蛋白尿患者中未发现ApoM。目前尚不清楚血浆鞘脂代谢的变化是如何导致糖尿病肾病的，以及个体脂蛋白类别在其中起什么作用

DESCRIPTION (provided by applicant): Sphingolipids have emerged as key signaling molecules involved in the regulation of a variety of cellular functions. Most research investigating sphingolipids has focused on the intracellular signaling functions of two bioactive sphingolipids - ceramide and S1P, but little attention has been given to the source of delivery of these molecules to cells. The proposed studies will focus on the extracellular transport of not only these sphingolipids but also that of sphingomyelin and the complex hexosyl- and lactosylceramide molecular species in plasma, urine, and in isolated lipoproteins and additionally, determine the changes in their transport which are associated with diabetes and nephropathy. The lipidomics profile of plasma obtained from 633 type 1 diabetic patients at their entry into the Diabetes Control and Complications Trial (DCCT), demonstrated that low plasma concentrations of several ceramide species predicted the development of macroalbuminuria. In contrast, the levels of sphingosine-1-phosphate (S1P), although 2.6-fold higher in diabetic patients compared to matched non-diabetic subjects, did not. Plasma concentrations of apolipoprotein M (ApoM), a physiological carrier of S1P, were significantly increased in Type 1 diabetic patients with albuminuria and ApoM was found in urine from these patients but not in normoalbuminuric patients. It is not known how changes in plasma sphingolipid metabolism contribute to nephropathy in diabetes and what role the individual lipoprotein classes play in this metabolism. We will test the hypothesis that plasma lipoproteins from type 1 diabetes patients, but especially HDL, are markedly enriched in S1P but poor in selected ceramides. Additionally, the decreased content of selected ceramides in HDL leads to their decreased concentrations both in plasma and urine. Lastly, measuring the levels of these ceramides in type 1 diabetes will identify patients at high risk to develop nephropathy at different stages of the disease. To evaluate these hypotheses, we will conduct studies with three Specific Aims: 1) Determine whether the levels of sphingolipids and ApoM measured in the plasma collected at the beginning and at the end of the DCCT trial in a subgroup of patients are associated with the development and progression of diabetic nephropathy; 2) Compare the sphingolipid and ApoM concentrations in plasma and urine from matched Type 1 diabetes patients with normoalbuminuria with those with macroalbuminuria, with non-diabetic subjects with macroalbuminuria, and with healthy controls; 3) Determine if the observed differences in plasma total sphingolipid and ApoM concentrations which are associated with diabetes and albuminuria are localized to VLDL, LDL or HDL. Knowledge of plasma sphingolipid transport in diabetes and in nephropathy will direct our future studies investigating the potential for lipoprotein sphingoliid to alter cell metabolism, especially cells associated with diabetic kidney disease.