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，偏头痛是一种 神经系统疾病，伴有使人丧失能力的间歇性头痛。TRPV 4 Ca++渗透性离子通道是 在三叉神经（TG）感觉神经元中，也在角质形成细胞和肥大细胞中表达和起作用， 两者都由头面部皮肤和脑膜中的TG感觉神经元支配。我的主要目标是定义TRPV 4 非神经元细胞，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 这些可以调节偏头痛相关的三叉神经伤害感受器神经元。一个特别有趣的和未被研究的 偏头痛和其他病理性疼痛的一个特点是种族差异的痛苦，确定为更严重的非洲 美国人理性理解的一个障碍是缺乏阐明这一点的机械研究 现象基于我的初步实验，即TRPV 4通道变得UVB敏感更多， 在非裔美国人皮肤角质形成细胞中， 在最前线三叉神经痛信号依赖于TRPV 4在角质形成细胞和肥大细胞。我的中枢 假设是皮肤角质形成细胞和脑膜肥大细胞中TRPV 4通道超敏 有助于偏头痛中炎性疼痛的发展和持续，并且这种调节是 受到种族背景的影响为了验证这一假设，我提出了两个目标：1）确定炎症是如何 改变角质形成细胞中TRPV 4通道的敏感性以及皮肤色素沉着如何决定通道的 2）研究TRPV 4在偏头痛中肥大细胞中的作用。从这个提议中屈服 将填补理解TRPV 4如何促进角质形成细胞和肥大细胞的串扰的重大空白， TG神经元，支配它们，并有助于偏头痛的病理生理学。解构这些 这一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。 痛觉器官功能对生物信号的可识别种族差异的探索， 询问细胞谱系将使我们能够利用这些发现走向个性化医疗。 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，我已经与他们进行了有效的合作，我建议为我的进一步发展进行有效的培训。 职业发展。我希望：1）获得合作和团队组建的经验，2）扩展我的技能- 在实验室为基础的技术，如动物行为和肥大细胞建模平台， 分子测定，3）获得并应用先进的统计分析技能，我的研究，4）扩大我的 沟通技巧，包括科学写作和演示，并建立在我现有的一套管理 和领导能力拟议的培训将使我具备成为一名 独立的实验室为基础的研究人员在多学科的方法研究炎症性疼痛。

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.