siRNA-gold nanoparticle mediated ganglioside depletion for diabetic wound healing

siRNA-金纳米粒子介导的神经节苷脂消耗促进糖尿病伤口愈合

基本信息

批准号：
8513708
负责人：
CHAD A. MIRKIN
金额：
$ 5.85万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2014-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8513708
关键词：
Address Adipocytes Cells Chronic Clinical Complication Contracture Cutaneous DNA Data Defect Diabetes Mellitus Diabetic mouse Diabetic wound Diet EGF gene Epidermal Growth Factor Receptor Event Foundations G(M3) Ganglioside Ganglioside Biosynthesis Pathway Gangliosides Gene Mutation Genes Genetic Genetic Suppression Glucose Glycosphingolipids Goals Gold Growth Factor Growth Factor Receptors Healed Histologic Human Hyperglycemia Impaired wound healing In Vitro Inbred HRS Mice Individual Infiltration Insulin Insulin Receptor Insulin Resistance Insulin-Like Growth Factor I Insulin-Like-Growth Factor I Receptor Integrins Intervention Laboratories Lead Ligands Link Measures Mediating Mediator of activation protein Membrane Modeling Molecular Mus Nanoconjugate Nanotechnology Non-Insulin-Dependent Diabetes Mellitus Obesity Oligonucleotides Phase I Clinical Trials Property Receptor Activation Receptor Signaling Research Role Safety Simulate Skin Small Interfering RNA Splint Device System Techniques Technology Testing Therapy Clinical Trials Thick Topical application United States Wound Healing base cell motility clinical application diabetic diabetic wound healing efficacy testing experience haematoside synthetase healing high risk improved inhibitor/antagonist innovation keratinocyte macrophage migration mouse model nano nanoparticle neovascularization nerve supply novel nucleic acid inhibitor pre-clinical prevent response success uptake wound

项目摘要

DESCRIPTION (provided by applicant): Improved management of wound healing represents a significant unmet need in the United States, particularly in individuals with obesity and type 2 diabetes. In addition, the molecular events that lead to insulin resistance remain poorly understood. Recent studies suggest that ganglioside GM3, a sialylated membrane glycosphingolipid, is a critical mediator of insulin resistance, as evidenced by the reversal of insulin resistance following ganglioside depletion in cultured adipocytes and diabetic mouse models. We have discovered that GM3 accumulates in keratinocyte (KC) membranes in diabetic mice, and that depletion of GM3 reverses their wound healing defect. We propose that genetic inhibition of ganglioside synthesis through the use of a novel nanotechnology approach will reverse impaired wound healing in KCs under high glucose conditions and in diabetic mice. The long-term goals of this project are to apply gene-suppressing topically-applied nanoparticles that block ganglioside biosynthesis as a new means to address the impaired wound healing in diabetics and to better understand how gangliosides impact KC proliferation and motility. We will use our unique oligonucleotide-conjugated gold nanoparticles (Au NPs), single agents that show universal uptake in cells and highly efficient gene knockdown. We will first evaluate the efficacy and safety of topically-applied GM3 synthase siRNA-Au NPs, which deplete gangliosides, in accelerating healing in diabetic mouse models. Next, we will determine how gangliosides impact KC motility. Using DNA- and siRNA-Au NPs to increase and deplete ganglioside GM3, respectively, we will assess KC proliferation and wound closure in vitro. We will then examine the effect of gangliosides on insulin receptor (IR), insulin-like growth factor-1 receptor-integrin (IGF- 1R), and epidermal growth factor receptor (EGFR) activation, all of which impact KC wound healing. Finally, we will evaluate the impact of GM3 depletion on glucose-induced insulin resistance. These studies will increase our understanding of the role of glycosphingolipids in wound healing. In addition, reversal of the wound healing defect in obese diabetic mice by topical administration of our nanoparticle-conjugated nucleic acid inhibitors of ganglioside synthesis will be an innovative means to promote wound healing in chronic wounds. These studies promise to have great impact in the treatment of wounds in humans, particularly in individuals with insulin-resistant diabetes.

描述（由申请人提供）：改善伤口愈合管理在美国是一个未满足的重大需求，特别是对于肥胖和 2 型糖尿病患者。此外，导致胰岛素抵抗的分子事件仍然知之甚少。最近的研究表明，神经节苷脂 GM3（一种唾液酸化膜鞘糖脂）是胰岛素抵抗的关键介质，培养的脂肪细胞和糖尿病小鼠模型中神经节苷脂耗尽后胰岛素抵抗的逆转就证明了这一点。我们发现 GM3 在糖尿病小鼠的角质形成细胞 (KC) 膜中积聚，并且 GM3 的消耗可以逆转其伤口愈合缺陷。我们提出，通过使用新型纳米技术方法对神经节苷脂合成进行基因抑制，将逆转高葡萄糖条件下的 KC 和糖尿病小鼠中受损的伤口愈合。该项目的长期目标是应用基因抑制局部应用纳米粒子来阻断神经节苷脂生物合成，作为解决糖尿病患者伤口愈合受损的新方法，并更好地了解神经节苷脂如何影响 KC 增殖和运动。我们将使用我们独特的寡核苷酸缀合金纳米颗粒 (Au NP)，这种单一药物可在细胞中显示出普遍吸收和高效的基因敲除。我们将首先评估局部应用的 GM3 合酶 siRNA-Au NP（可消耗神经节苷脂）在加速糖尿病小鼠模型愈合方面的功效和安全性。接下来，我们将确定神经节苷脂如何影响 KC 运动性。使用 DNA-和 siRNA-Au NP 分别增加和消耗神经节苷脂 GM3，我们将在体外评估 KC 增殖和伤口闭合。然后我们将检查神经节苷脂对胰岛素受体 (IR)、胰岛素样生长因子-1 受体整合素的影响 (IGF-1R) 和表皮生长因子受体 (EGFR) 激活，所有这些都会影响 KC 伤口愈合。最后，我们将评估 GM3 消耗对葡萄糖诱导的胰岛素抵抗的影响。这些研究将增加我们对鞘糖脂在伤口愈合中作用的理解。此外，通过局部施用我们的纳米颗粒缀合的神经节苷脂合成核酸抑制剂来逆转肥胖糖尿病小鼠的伤口愈合缺陷将是促进慢性伤口愈合的创新方法。这些研究有望对人类伤口的治疗产生巨大影响，尤其是胰岛素抵抗型糖尿病患者。