Sensory neuron-skin interaction in health and disease

项目摘要 我的职业目标是领导一个独立的研究小组，调查的病理和决议， 疼痛来解决神经性疼痛问题。为了实现这一目标，我建议开发和使用新的翻译 使用人类和动物的组合研究皮肤层中神经元-基质相互作用的策略 周围感觉神经病模型。我的研究集中在一种常见的外周感觉 化疗引起的神经病变，仅在美国就影响了超过300万人。化疗的时候- 诱导性周围神经病变（CIPN）在高达80%的用化疗剂治疗的癌症患者中发展， 没有有效的预防或治疗策略。考虑到细胞外环境的动态变化， 皮肤感觉神经和最近的理解，角质形成细胞也可以作为一个伤害感受器，它是 这对于理解神经元和角质形成细胞的维持和功能以解决神经性疼痛至关重要。 因此，我建议测试的假设，失调的神经元和表皮的相互作用， CIPN的病理进展。根据我最近的工作，确定了整合素的保护作用， CIPN，我的建议有三个目标，使用尖端工具和创新的多学科方法。在 目的1，在果蝇体内研究神经元-底物相互作用及其调控机制 CIPN模型我将使用遗传学和先进的显微镜方法来标记，操作和检测 CIPN模型中神经元与基质接触的亚细胞变化以及与功能变化的联系。使用 整合素作为一种模型，以了解细胞表面蛋白介导的机制，CIPN的基础上，我将 表征调节介导神经元-底物接触的表面蛋白表达的内吞调节剂。 目的2：利用人诱导多能干细胞（iPSC）技术，鉴定角朊细胞-伤害感受性细胞， 在共培养CIPN模型中的神经元相互作用和细胞类型特异性病理。由于物种间的差异， 感觉神经元，我希望建立一个人类模型来研究感觉神经元的影响， 外在因素对CIPN病理进展的影响将提供重要的见解。在目标3中，我将建立和 验证3D人类皮肤-神经共培养CIPN模型以研究神经元-基质关系， 整合素介导的保护。由于伤害感受神经元的亚型靶向不同的表皮层， 有助于周围神经病变的独特病理学，3D工程人类皮肤将被纳入 模型这将使我能够研究亚型特异性病理学，并解决伤害性神经元末端 CIPN的变性和功能障碍。该提案将为病理进展提供新的见解， 神经元基质相互作用介导的机制，为我未来的研究奠定了坚实的基础。作为我 主要专长是利用果蝇模型研究人类神经元和表皮细胞的神经科学 需要在皮肤生物学和干细胞生物学方面进行新的培训。这个计划对我的事业有很大的帮助 目标是解决患者顽固性神经性疼痛问题。

PROJECT SUMMARY My career goal is to lead an independent research group to investigate the pathology and resolution of pain to solve neuropathic pain problems. To achieve this goal, I propose to develop and use novel translational strategies to investigate neuron-substrate interactions in the skin layer using a combination of human and animal models of peripheral sensory neuropathy. My research focuses on a common type of peripheral sensory neuropathy induced by chemotherapy, which affects over 3 million people in the US alone. While chemotherapy- induced peripheral neuropathy (CIPN) develops in up to 80% of cancer patients treated with chemotherapeutics, there are no effective prevention or treatment strategies. Considering the dynamic extracellular environment of cutaneous sensory nerves and recent understanding that keratinocytes can also function as a nociceptor, it is vital to understand the maintenance and function of both neurons and keratinocytes to solve neuropathic pain. Hence, I propose to test the hypothesis that dysregulation of neuron and epidermal interactions underlie pathological progression of CIPN. Building from my recent work that identified protective roles of integrins in CIPN, my proposal has three aims that use cutting-edge tools and innovative multi-disciplinary approaches. In Aim 1, I will investigate neuron-substrate interactions and their regulatory mechanisms in a Drosophila in vivo CIPN model. I will use genetics and advanced microscopy approaches to label, manipulate, and detect subcellular changes in neuron to substrate contacts and link with functional changes in a CIPN model. Using integrins as a model to understand cell surface protein-mediated mechanisms underlying CIPN, I will characterize endocytic regulators that modulate surface protein expression mediating neuron-substrate contacts. In Aim 2, I will use human induced pluripotent stem cell (iPSC) technology to identify keratinocyte-nociceptive neuron interactions and cell-type-specific pathology in a co-culture CIPN model. Given species differences in sensory neurons, I expect that establishing a human model to investigate the impact of sensory neurons and extrinsic factors on pathological progression in CIPN will provide critical insights. In Aim 3, I will establish and validate a 3D human skin-nerve co-culture CIPN model to investigate neuron-substrate relationships and integrin-mediated protection. Because subtypes of nociceptive neurons target different epidermal layers and contribute to distinct pathology in peripheral neuropathy, a 3D engineered human skin will be incorporated in the model. This will allow me to investigate subtype-specific pathology and to resolve nociceptive neuron terminal degeneration and dysfunction in CIPN. This proposal will provide new insights into pathological progression and mechanisms mediated by neuron-substrate interactions and a strong foundation for my future research. As my primary expertise is in neuroscience using Drosophila models, investigating human neurons and epidermal cells requires new training in skin biology and stem cell biology. The proposed plan will significantly facilitate my career goal to solve intractable neuropathic pain problems in patients.