Neuron-Keratinocyte Communication in the Epidermis in Normal and Diabetic Wound Healing

正常和糖尿病伤口愈合中表皮神经元-角质形成细胞的通讯

• 批准号: 10472010
• 负责人: Daniela M Menichella
• 金额: $ 66.21万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-11 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10472010
• 关键词: 3-Dimensional; Agonist; American; Behavior; Calcium Signaling; Cell Proliferation; Cells; Chronic; Communication; Cutaneous; Data; Diabetes Mellitus; Diabetic mouse; Drug Targeting; Epidermis; Fiber; G alpha q Protein; G-Protein-Coupled Receptors; Gene Expression Profile; Genes; Genetic; Genetic Techniques; Goals; Health; Human; Impaired healing; Impaired wound healing; Individual; Insulin Resistance; Intervention; Lead; Link; Molecular; Morbidity - disease rate; Mus; Nerve; Nerve Degeneration; Nerve Regeneration; Neurons; Neurotransmitters; Non-Insulin-Dependent Diabetes Mellitus; Pharmaceutical Preparations; Pharmacology; Play; Population; Regulation; Role; Sensory; Signal Transduction; Skin; Spinal Ganglia; Splint Device; Subgroup; Testing; Topical application; Toxic effect; Type 2 diabetic; Wound models; afferent nerve; base; cell motility; cell type; cutaneous sensory nerve; designer receptors exclusively activated by designer drugs; diabetic; diabetic ulcer; diabetic wound healing; diphtheria toxin receptor; drug development; drug discovery; genetic manipulation; healing; high throughput screening; high throughput technology; improved; keratinocyte; migration; molecular marker; mouse model; non-diabetic; novel therapeutic intervention; novel therapeutics; painful neuropathy; preclinical trial; prevent; receptor; skin ulcer; small molecule; transcriptome sequencing; transcriptomics; wound; wound bed; wound healing

Project Summary/Abstract: Poor wound healing is a major health issue in insulin-resistant diabetes. Degeneration of nerves in diabetes contributes to the delay in healing and is associated with reduction in basal keratinocyte migration across the wound bed. Improved understanding of the communication between neurons and keratinocytes, which is critical for wound repair, may lead to new interventions. Cutaneous sensory nerves are now recognized to comprise several subtypes characterized by different markers and functions. Identifying the neuron subtype(s) involved in wound healing may provide clues to new therapeutic directions. To explore the impact of specific neuron-keratinocyte communication on wound healing, we will initially ablate specific neuron subsets in healthy mouse skin using genetic expression of diphtheria toxin receptors and will evaluate the impact on healing of splinted wounds. We will confirm subgroup neurons involvement by chemogenetically introducing and activating stimulatory designer receptors (DREADDs), which we expect to accelerate healing if activated in a nerve subset that is important for normal healing. We have shown that nerve degeneration results from neuronal hyperexcitability and that introducing inhibitory DREADDs into the majority of sensory nerves in a mouse model of diabetes both suppresses this excitation and reverses the nerve degeneration, although the impact on healing is unexplored. Building on this observation, we will introduce these inhibitory DREADDs into specific neuronal subsets in diabetic mice to delineate the impact on healing and whether one or more subtype of neurons is key to the degeneration and healing impairment. These studies in healthy and diabetic mice will allow us to capture unwounded and wound edge skin for conducting transcriptomic analysis. In particular, we will evaluate changes in expression of G-protein coupled receptors (GPCRs). Activation of these GPCRs with selective agonists should replicate the observed effects of DREADDs. We anticipate that these studies will implicate targets in keratinocytes for small molecule drug discovery using high throughput technology to assess calcium signals, migration, proliferation, and toxicity. Best candidates will be tested topically in cultured 3D human diabetic wound models and subsequently in our type 2 diabetic mouse models towards finding new interventions to promote wound healing. These proposed studies will increase our understanding of the role that nerve afferent subsets play in diabetic vs. normal wound healing. Furthermore, by identifying responsible subsets of nerves and gene expression patterns that are altered during diabetic wound healing, we can screen and advance preclinical trials of new small molecules that can be applied topically to promote healing of diabetic wounds.

项目摘要/摘要： 伤口愈合不佳是耐胰岛素糖尿病的主要健康问题。糖尿病中神经的变性 有助于延迟愈合，并与跨基底角质形成细胞迁移的减少有关 伤口床。对神经元与角质形成细胞之间的沟通的了解得以提高，这是 对于伤口维修至关重要，可能会导致新的干预措施。现在认识到皮肤的感觉神经 包括以不同标记和功能为特征的几种亚型。识别神经元亚型 涉及伤口愈合可能会为新的治疗方向提供线索。探索特定的影响 神经元 - 甲状腺细胞在伤口愈合方面的通信，我们最初将在健康中烧毁特定的神经元子集 使用白喉毒素受体的遗传表达的小鼠皮肤，将评估对愈合的影响 夹住伤口。我们将通过化学引入和 激活刺激性设计器受体（Dreadds），如果在A中激活，我们希望它会加速愈合 对于正常愈合很重要的神经子集。我们已经表明，神经变性来自 神经元的过度兴奋性，并将抑制性恐怖引入大多数感觉神经 糖尿病的小鼠模型都抑制了这种激发并逆转神经变性，尽管 对治疗的影响尚未探索。在这一观察结果的基础上，我们将向这些抑制性恐怖介绍 糖尿病小鼠中的特定神经元亚群，以描述对愈合的影响以及一种或多种亚型 神经元是退化和治愈障碍的关键。这些在健康和糖尿病小鼠中的研究将 允许我们捕获未能的和伤口的边缘皮肤进行转录组分析。特别是我们 将评估G蛋白偶联受体（GPCR）表达的变化。这些GPCR激活 选择性激动剂应复制恐惧的观察到的影响。我们预计这些研究将 在角质形成细胞中暗示了使用高吞吐量技术发现小分子药物的靶标 评估钙信号，迁移，增殖和毒性。最好的候选人将在培养中局部测试 3D人类糖尿病伤口模型，随后在我们的2型糖尿病小鼠模型中找到新的 干预措施以促进伤口愈合。这些提出的研究将增加我们对角色的理解 那个神经传入的子集在糖尿病和正常伤口愈合中发挥作用。此外，通过确定负责任 糖尿病伤口愈合期间改变的神经和基因表达模式的子集，我们可以筛查 并预先对新的小分子进行临床前试验，可以局部应用 糖尿病伤口。