Glycosphingolipids Mediate Diabetic Wound Healing Impairment

鞘糖脂介导糖尿病伤口愈合受损

• 批准号: 9898159
• 负责人: Amy S Paller
• 金额: $ 40.31万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9898159
• 关键词: 3-Dimensional; Acceleration; Acids; Address; Age; Amputation; Attention; Binding; Biochemical; Blood Vessels; Cell Nucleus; Cell membrane; Ceramide glucosyltransferase; Chronic; Clathrin; Coculture Techniques; Cutaneous; Data; Defect; Diabetes Mellitus; Diabetic Diet; Diabetic Foot; Diabetic mouse; Diet; Dose; Endocytosis; Engineering; Enzymes; Epithelial; Epithelium; Fibroblasts; Fluorescence; Fluorescence Resonance Energy Transfer; G(M3) Ganglioside; Gangliosides; Gene Delivery; Gene Expression; Gene Expression Regulation; Genes; Genetic; Glucose; Glycolipids; Glycosphingolipids; Goals; Growth Factor; Health; Histologic; Human; Hybrids; Hyperglycemia; Impaired healing; Impaired wound healing; Individual; Infection; Infiltration; Insulin; Insulin Resistance; Insulin-Like Growth Factor I; Insulin-Like-Growth Factor I Receptor; Intervention; Knock-out; Mediating; Membrane; Microbial Biofilms; Microscopy; Modeling; Morbidity - disease rate; Mus; Nano delivery; Nanoconjugate; Neuropathy; Obese Mice; Obesity; Oral; Pathology; Pathway interactions; Pharmacology; Pre-Clinical Model; Process; Receptor Activation; Receptor Signaling; Resolution; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Skin; Small Interfering RNA; Spherical Nucleic Acids; Splint Device; TNF gene; Testing; Therapeutic; Tomatoes; Topical application; Vascular Diseases; Western Blotting; Wound models; afferent nerve; base; blood glucose regulation; chronic wound; comparison intervention; crosslink; cytokine; db/db mouse; diabetic; diabetic ulcer; diabetic wound healing; epithelial wound; haematoside synthetase; healing; improved; in vivo; inhibitor/antagonist; intercellular communication; keratinocyte; keratinocyte growth factor receptor; macrophage; migration; nanoparticle; nerve supply; novel; prevent; public health relevance; receptor; response; small molecule; therapeutic evaluation; wound; wound biofilm; wound closure; wound healing

 DESCRIPTION (provided by applicant): Poor wound healing is a major health issue in insulin-resistant diabetes. Improved understanding of wound pathology, including the function of keratinocytes (KCs) in re-epithelialization, and new interventions for impaired wound healing are needed. Ganglioside GM3 is a glycosphingolipid that regulates receptor signaling at the membrane level. We have found that GM3 and GM3 synthase (GM3S) are increased in human and mouse diabetic skin. In addition, TNF and excess glucose-induced insulin resistance in KCs is associated with increased GM3 and reversed by GM3 depletion, suggesting that GM3 mediates diabetic wound healing impairment. Wounds heal normally in diet-induced obese (DIO) GM3 synthase knockout (GM3S-/-) mice, in contrast to delayed healing in DIO wildtype (WT) littermates. Furthermore, we have replicated reversal of this wound healing improvement in WT DIO diabetic mice by depleting GM3 with topically applied GM3S siRNA spherical nuclei acid (SNA) nanoparticle conjugates. Depleting GM3 by either GM3S SNA or GZ 161, a glucosylceramide synthase inhibitor, accelerates migration in 2D keratinocyte cultures by activating insulin-like growth factor-1 receptor (IGF1R) and Rac1 signaling. Increases in GM3, including by treatment with excess glucose or chronic, low-dose TNF, suppress IGF1R signaling and inhibit KC migration. Our long-term goals are to test GM3 depletion in skin as a novel means to reverse the impaired wound healing in diabetics and understand how GM3 inhibits IGF1R activation and suppresses keratinocyte migration. Our first aim is to optimize GM3 depletion-based intervention by comparing the ability of topical GM3S siRNA SNA and topical or oral GZ 161 to promote healing. We will test these therapeutic options in a DIO mouse with fluorescent sensory nerves to track the impact of treatment on cutaneous innervation. The most efficacious therapy will then be studied in a diabetic model with more chronic wounds, the biofilm-challenged db/db mouse. Using normal and diabetic 3D co-culture wound models, we will validate our observations in 2D KCs and will investigate the effects of GM3 modulation on the insulin/IGF-1 signaling axis (Aim 2). Finally, as our third aim and with a focus on IGF1R, we will elucidate the role of GM3 in regulating the membrane-based localization, dynamics, and molecular interactions of insulin/IGF-1 signaling. We will interrogate the direct interactions of IGF1R with GM3 and test our hypothesis that increased GM3 prevents IGF1R clathrin-mediated endocytosis and signaling. Finally, we will use FLIM to determine if GM3 depletion increases IR-IGF1R hybrid receptors to boost IGF-1 responses. These proposed studies will increase our understanding of the membrane-based dynamics that impact insulin/IGF-1 signaling in skin. Furthermore, acceleration of wound healing, whether by nano-delivery of gene suppression or small molecule inhibition of GM3 synthesis, could be fast-tracked towards translational application as a new treatment approach for diabetic wounds.

 描述（由申请人提供）：池伤口健康是一种胰岛素的糖尿病。受体信号在人类和小鼠糖尿病的皮肤中。肥胖（DIO）GM3合酶基因敲除（GM3S - / - ）小鼠，与Dio WildType（WT）胆小号的延迟相比，我们通过局部应用GM3耗尽了GM3的GM3 Sirna，使这种伤口愈合的逆转复制了这种伤口愈合球酸（SNA）纳米颗粒结合物，通过GM3S SNA或GZ 161耗尽GM3，一种葡萄糖基酶合酶抑制剂，2D角质形成细胞培养，通过激活胰岛素样生长因子（IGF1R）和RAC1信号，或慢性剂量TNF抑制IGF1R信号和抑制KC迁移。口服GZ 161促进疗程。挑战的DB/DB小鼠在2D KC中的观察结果将研究GM3调制对胰岛素/IGF -1信号轴的影响（AIM 2）胰岛素/IGF-1信号传导。为了引导IGF-1反应。可以快速跟踪转化应用，作为一种糖尿病伤口的新治疗方法。

项目成果

Abnormal scar identification with spherical-nucleic-acid technology.

• DOI: 10.1038/s41551-018-0218-x
• 发表时间: 2018-04
• 期刊: Nature biomedical engineering
• 影响因子: 28.1
• 作者: Yeo DC;Wiraja C;Paller AS;Mirkin CA;Xu C
• 通讯作者: Xu C