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

• 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
• 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

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 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表现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，特别是胰岛素抵抗性糖尿病患者。