Localized Vulvodynia Pathogenesis: Fibroblast, Yeast, and Melanocortin

描述（由申请人提供）：本R01申请响应PAR-10-190：“外阴痛-系统流行病学，病因学或治疗研究”。我们的长期目标是发展外阴痛疼痛机制的理解，导致基于机制的疾病分类，并最终以机制为基础的治疗。我们的研究小组已经报道了酵母产物、局部成纤维细胞激活、促肾上腺素和局部诱发性外阴痛（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作出反应，而且可以大量产生许多不同的生物介质，包括那些促进疼痛的生物介质。成纤维细胞也表现出相当大的区域特化。我们发现来自外阴前庭的成纤维细胞在酵母细胞壁产物激活后产生显著升高的促炎、促痛介质水平。特别是，来自lpv患者外阴前庭的成纤维细胞产生了增强的促炎介质反应。这可能与黑色素皮质素-1受体（MC1R）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/疼痛诱导反应性，外阴痛痛反映了一种极端但自然的炎症现象。我们假设外阴痛产生于1)易受炎症影响的生殖道区域，2)特定刺激物（如酵母菌）的存在，3)遗传易感性加剧了外阴痛。为了显著推进和影响该领域，我们组建了一个在LPV，成纤维细胞生物学和炎症方面经验丰富的多学科团队，以实现以下三个目标。特异性目的1：确定促炎成纤维细胞是否分离到外阴疼痛区域。使用下生殖道疼痛制图，我们将发现促炎成纤维细胞是否定位于疼痛的原位解剖区域。成纤维细胞菌株将从外阴疼痛和非疼痛区域发展，其对促炎和其他介质的生物合成能力在暴露于关键的成纤维细胞激活因子后决定。特异性目的2：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，以及特定的MC1R snp是否改变这种反应。我们将确定lpv患者是否携带不同的酵母种类和酵母负荷模式，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、疼痛诱导反应。特异性目的3：确定促肾上腺素“功能丧失”是否促进外阴痛。我们将研究“功能缺失”的黑素皮质素-1受体snp是否会增强位点特异性成纤维细胞的激活，并且可以通过一种简单的临床测量方法——皮肤比色法来识别。我们将评估一种具有治疗外阴痛潜力的抗炎黑素皮质素衍生物，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).