Localized Vulvodynia Pathogenesis: Fibroblast, Yeast, and Melanocortin

描述(由申请人提供)：此R01申请响应PAR-10-190：“外阴疼痛--系统性流行病学、病因学或治疗学研究”。我们的长期目标是发展对外阴疼痛机制的理解，导致基于机制的疾病分类，并最终实现基于机制的治疗。我们的研究小组报告了酵母产物、区域成纤维细胞激活、前阿片黑素皮质素和局限性外阴疼痛(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地区性特化。我们发现，外阴前庭的成纤维细胞在酵母细胞壁产品激活后，会产生明显升高的促炎、促痛介质水平。特别是，患有LPV的女性外阴前庭成纤维细胞产生了高度的促炎介质反应。这可能与黑素皮质素-1受体(MC1R)的单核苷酸多态性(SNP)有关，该基因可促进炎症介质的产生。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，而外阴疼痛反映了一种极端但自然的炎症现象。我们假设外阴疼痛发生在1)容易发炎的生殖道区域，2)在酵母等特定刺激物的存在下，3)遗传易感性加剧。为了显著推进和影响该领域，我们组建了一支在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。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。使用下生殖道疼痛标测，我们将发现促炎成纤维细胞是否定位于原位疼痛的解剖区域。成纤维细胞株将从外阴疼痛和非疼痛区域培养出来，并在暴露于关键的成纤维细胞激活细胞因子后确定其对促炎和其他介质的生物合成能力。具体目标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是否改变这一反应。我们将确定LPV患者是否携带不同模式的酵母种类和酵母负荷，以及酵母细胞壁产品是否通过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。具体目标3：确定前阿片黑素皮质素“功能丧失”是否促进外阴疼痛。我们将研究“功能丧失”的黑素皮质素-1受体SNPs是否能促进特定部位成纤维细胞的激活，并可用一种简单的临床测量方法--皮肤比色法进行鉴定。我们将评估一种具有治疗外阴疼痛潜力的抗炎黑素皮质素衍生物，并探讨其潜在的分子机制(S)。

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).