Localized Vulvodynia Pathogenesis: Fibroblast, Yeast, and Melanocortin

描述（由申请人提供）：本R 01申请响应PAR-10-190：“外阴痛-系统流行病学、病因学或治疗研究。“我们的长期目标是了解外阴疼痛的机制，从而导致基于机制的疾病分类，并最终导致基于机制的治疗。我们的研究小组报告了酵母产物、局部成纤维细胞活化、前阿黑皮素和局部激发性外阴痛（LPV）之间的机制联系。成纤维细胞现在被认为不仅仅是结构细胞，因为它们不仅对信号做出反应，而且可以产生许多不同的生物介质，包括那些促进疼痛的介质。成纤维细胞也表现出相当大的区域性特化。我们发现，来自外阴前庭的成纤维细胞在用酵母细胞壁产物活化后产生显著升高水平的促炎、促痛介质。特别是，增强的促炎介质的反应是由成纤维细胞从外阴前庭的LPV折磨的妇女。这可能与黑皮质素-1受体（MC 1 R）中的单核苷酸多态性（SNP）有关，该多态性可增强炎症介质的产生。我们认为所有女性的外阴前庭都具有独特的炎症/疼痛诱导反应，外阴疼痛反映了一种极端但自然的炎症现象。我们假设外阴痛发生在1）生殖道易患炎症的区域，2）存在特定刺激物（如酵母）时，3）因遗传易感性而加剧。为了显著推进和影响该领域，我们组建了一个多学科团队，在LPV，成纤维细胞生物学和炎症方面经验丰富，以实现以下三个目标。具体目标1：确定促炎成纤维细胞是否分离到外阴疼痛区域。使用下生殖道疼痛映射，我们将发现促炎成纤维细胞是否定位于原位疼痛解剖区域。将从外阴的疼痛和非疼痛区域开发成纤维细胞株，并在暴露于关键成纤维细胞活化细胞因子后确定其促炎和其他介质的生物合成能力。具体目标二：确定酵母或酵母产品是否通过Toll样受体（TLR）激活成纤维细胞，以及特定MC 1 R SNP是否改变该反应。我们将确定LPV患者是否携带不同的酵母菌种类和酵母菌负荷模式，以及酵母细胞壁产物是否通过Toll样受体启动来自疼痛区域的成纤维细胞的促炎性疼痛诱导反应。具体目标3：确定阿黑皮素原“功能丧失”是否促进外阴痛。我们将研究是否“功能丧失”黑皮质素-1受体单核苷酸多态性增强位点特异性成纤维细胞活化，并可以确定一个简单的临床措施，皮肤比色法。我们将评估一种具有治疗外阴痛潜力的抗炎黑皮质素衍生物，并研究其潜在的分子机制。

DESCRIPTION (provided by applicant): This R01 application responds to PAR-10-190: "Vulvodynia - Systematic Epidemiologic, Etiologic or Therapeutic Studies." Our long-term goal is to develop an understanding of the vulvodynia pain mechanism leading to a mechanism-based disease classification and ultimately to a mechanism-based therapy. Our research team has reported a mechanistic connection between yeast products, regional fibroblast activation, pro-opiomelanocortins, and localized provoked vulvodynia (LPV). Fibroblasts are now recognized as more than structural cells as they not only respond to signals but can prodigiously produce many different biologic mediators, including those that promote pain. Fibroblasts also exhibit considerable regional specialization. We discovered that fibroblasts from the vulvar vestibule produce markedly elevated levels of pro-inflammatory, pro-pain mediators following activation with yeast cell wall products. In particular, heightened pro-inflammatory mediator responses are generated by fibroblasts from the vulvar vestibule of LPV-afflicted women. This may be related to single nucleotide polymorphisms (SNP) in the melanocortin-1 receptor (MC1R) that enhance inflammatory mediator production. We propose that the vulvar vestibule of all women possesses a unique inflammatory/pain-inducing responsiveness and that vulvodynia pain reflects an extreme but natural inflammatory phenomenon. We hypothesize that vulvodynia arises 1) in a region of the genital tract predisposed to inflammation, 2) in the presence of specific irritants such as yeast, that are 3) exacerbated by genetic predisposition. To significantly advance and impact the field, we have assembled a multidisciplinary team, experienced in LPV, fibroblast biology, and inflammation to achieve the following three aims. Specific Aim 1: To determine whether pro-inflammatory fibroblasts segregate to painful areas of the vulva. Using lower genital tract pain mapping, we will discover whether pro-inflammatory fibroblasts localize to painful anatomic regions in situ. Fibroblast strains will be developed from painful and non-painful areas of the vulva and their biosynthetic capabilities for pro-inflammatory and other mediators determined after exposure to key fibroblast activating cytokines. Specific Aim 2: To determine whether yeast or yeast products activate fibroblasts via Toll-like receptors (TLR) and whether specific MC1R SNPs modify that response. We will determine whether the LPV-afflicted patients carry a different pattern of yeast species and yeast load, and whether yeast cell wall products initiate, through toll-like receptors, a pro-inflammatory, pain-inducing response from fibroblasts derived from painful regions. Specific Aim 3: To determine whether pro-opiomelanocortin "loss-of-function" promotes vulvodynia. We will investigate whether "loss-of-function" melanocortin-1 receptor SNPs enhance site-specific fibroblast activation, and can be identified with a simple clinical measure, skin colorimetry. We will assess an anti-inflammatory melanocortin derivative with therapeutic potential for vulvodynia and investigate the underlying molecular mechanism(s).