Molecular and Cellular signaling mechanisms of TRPV4 in non-neuronal cells: a pathophysiological relevance for migraine pain.

摘要 瞬时受体电位(Trp)离子通道与偏头痛的病理生理学有关。 伴有阵发性头痛的神经性疾病。TRPV4钙离子渗透性离子通道是 在三叉神经(TG)感觉神经元、角质形成细胞和肥大细胞中表达和功能 两者均由TG感觉神经元支配，见于头面部皮肤和脑膜。我的主要目标是定义TRPV4 非神经细胞，特别是皮肤角质形成细胞和脑膜肥大细胞的依赖机制以及如何 这些神经元可以调节偏头痛相关的三叉神经痛感受器神经元。一个特别耐人寻味但未被充分研究的问题 偏头痛和其他病理性疼痛的特征是痛苦的种族差异，在非洲被认为更严重 美国人。理性理解的障碍仅仅是缺乏阐明这一点的机械性研究。 现象。根据我的初步实验，即TRPV4频道对UVB更敏感 非裔美国人的皮肤角质形成细胞比高加索人的角质形成细胞，我也打算阐明种族差异。 三叉神经前部疼痛信号依赖于角质形成细胞和肥大细胞中的TRPV4。我的中央 假设TRPV4通道在皮肤角质形成细胞和脑膜肥大细胞中的超敏反应 有助于偏头痛炎症性疼痛的发展和永久存在，这一调节是 受种族背景的影响。为了验证这一假设，我提出了两个目标：1)确定炎症如何 改变角质形成细胞中TRPV4通道的敏化以及皮肤色素沉着如何决定通道 2)研究TRPV4在偏头痛中肥大细胞中的作用。从这项提议中获得收益 将填补在理解TRPV4如何促进角质形成细胞和肥大细胞与 三叉神经节神经元支配它们，参与偏头痛的病理生理学。解构这些 这些过程将为调节肥大细胞和角质形成细胞强大的促炎/促炎作用奠定基础。 代数有机体函数。提出的对生物信号的可识别种族差异的探索 被询问的细胞谱系将使我们能够利用这些发现走向个性化医学。 在我的导师团队的指导下，他们的专业知识涵盖了相关的研究领域 对于这项提议，以及我已经与之有效合作的人，我提议有效地培训我的进一步 职业发展。我希望：1)在协作和团队组建方面获得经验，2)扩大我的技能- 建立基于实验室的技术平台，如动物行为建模和肥大细胞和 分子分析，3)获得高级统计分析技能并将其应用于我的研究，4)扩展我的 沟通能力，包括科学写作和演示，并建立在我现有的一套管理 和领导才能。拟议的培训将使我掌握一套独特的技能，以便成为一名 研究炎性疼痛的多学科方法的独立实验室调查员。

ABSTRACT Transient receptor potential (TRP) ion channels have been implicated in the pathophysiology of migraine, a neurological disorder with incapacitating episodic headaches. TRPV4 Ca++-permeable ion channels are expressed and function in trigeminal (TG) sensory neurons, also in keratinocytes and in mast cells, which are both innervated by TG sensory neurons, in head-face skin and meninges. My main objective is to define TRPV4 dependent mechanisms in non-neuronal cells, specifically skin keratinocytes and meningeal mast cells, and how these can tune migraine pain-relevant trigeminal nociceptor neurons. One particular intriguing and understudied feature of migraine and other pathologic pain is racial divergence of suffering, identified as more severe in African Americans. An obstacle toward rational understanding is simply a lack of mechanistic studies that elucidate this phenomenon. Based on my preliminary experiments, namely TRPV4 channels becoming UVB-sensitized more in skin keratinocytes from African Americans than from Caucasians, I also intend to elucidate racial differences in forefront trigeminal pain signaling that depend on TRPV4 in keratinocytes and mast cells. My central hypothesis is that hypersensitization of TRPV4 channels in skin keratinocytes and meningeal mast cells contributes to development and perpetuation of inflammatory pain in migraine, and that this regulation is modulated by racial background. To test this hypothesis, I propose two aims: 1) To determine how inflammation alters TRPV4 channel sensitization in keratinocytes and how skin pigmentation determines the channel’s sensitivity; 2) To characterize the contribution of TRPV4 in mast cells in migraine pain. Yield from this proposal will fill a significant gap in understanding how TRPV4 facilitates the crosstalk of keratinocytes and mast cells with TG neurons that innervate them and contribute to the pathophysiology of migraine. Deconstructing these processes will set the stage for regulating both mast cells’ and keratinocytes’ powerful pro-inflammatory/pro- algesic organismal function. The proposed quest for identifiable racial differences of biologic signaling in the interrogated cell lineages will position us to leverage these findings toward personalized medicine. With the guidance from my team of mentors, whose expertise ranges across the pertinent areas of study relevant for this proposal, and with whom I am already effectively collaborating, I propose to effectively train for my further career development. I expect to: 1) Gain experience in collaborations and team formation, 2) Expand my skill- set platform in laboratory-based techniques such as modeling animal behavior and mast cell cellular and molecular assays, 3) Acquire and apply advance statistical analysis skills to my research, 4) Expand my communication skills, including scientific writing and presentation, and build on my existing set of managerial and leadership skills. The proposed training will equip me with unique a set of skills needed to become an Independent laboratory-based investigator in multidisciplinary approaches for studying inflammatory pain.