Collagen XVII unmasking in Parkinson's disease and scratching

项目总结 大疱性类天疱疮(BP)是一种自身免疫性疾病，会导致严重的瘙痒和水泡。 它是最常见的自身免疫性水泡病，在老年人中表现过度。治疗 对于大疱性类天疱疮依赖于免疫抑制的使用，这在医学上有很高的患病率 在这一组中，合并疾病导致一年死亡率从13%到27%不等。大疱性类天疱疮 由抗XVI型胶原抗体的产生导致水泡形成和 炎症的发展。丧失对XVII胶原蛋白的自身耐受性，特别是在老化的免疫系统中，是 被认为是导致BP发展的原因。帕金森病和既往的瘙痒症都是独立的 对大疱性类天疱疮的发展有强烈的易感性，可能是通过自身接种 含有Xvii胶原蛋白。了解XVII胶原自身接种的机制不仅提供了洞察力 大疱性类天疱疮的发病机制，但提供了对免疫反应的重要洞察力 帕金森氏症。我们提出了两个离散的假设：1)抓挠导致皮肤屏障损伤或 2)帕金森病患者出现的神经炎症足以在耐受的情况下自体接种XVII胶原 最低T细胞耐受性。为了验证这一假设，我们建议使用一种新的动物模型。我们已经使用了 CRISPR-Cas9开发了一种表达loxP-PolyA-loxP外显子18上游序列的小鼠，编码 XVII胶原蛋白NC15a结构域的区域。该结构域对应于人的NC16a结构域，该结构域 作为BP的主要自身抗原。当与他莫昔芬诱导的Cre-Lox小鼠模型杂交时， XVII胶原蛋白表达的恢复发生在成年早期，一旦暴露，就会绕过T细胞的耐受性。 通过抓挠或实验诱导的帕金森氏症，炎症和抗原暴露， 我们预计会出现抗原特异性的CD4+T细胞和抗XVI型胶原的自身抗体。二 提出了具体目标：1)确定划痕行为对抗划痕行为发展的影响 XVI型胶原自身抗体、抗原特异性的CD4+T细胞与BP的发生发展。2)确定 实验性帕金森病中神经炎症在诱导皮肤抗胶原自身免疫中的作用 第十七条。这个项目将提供对抓挠和神经炎症的作用的重要见解。 抗原揭开面纱。这一提议很可能在几种疾病上具有很强的翻译潜力。 过程和器官系统。我们的新方法为研究免疫反应提供了一个有价值的工具。 在帕金森氏病以及研究其他自身免疫性疾病方面具有很好的特点 自身抗原。

PROJECT SUMMARY Bullous pemphigoid (BP) is an autoimmune disease that results in the development of severe itch and blisters. It is the most common form of autoimmune blistering disease and is overrepresented in the elderly. Treatment for bullous pemphigoid relies on the use of immunosuppression, which given the high prevalence of medical comorbidities in this group, results in one-year mortality rates ranging from 13% to 27%. Bullous pemphigoid is caused by the development of antibodies against collagen XVII leading to blister formation and the development of inflammation. Loss of self-tolerance to collagen XVII, especially in the aging immune system, is thought to lead to development of BP. Parkinson's disease and pre-existing pruritus both independently impose strong predisposition towards the development of bullous pemphigoid, likely through autoinoculation with collagen XVII. Understanding the mechanism of collagen XVII autoinoculation not only provides insight into the pathogenesis of bullous pemphigoid, but offers significant insight into the immune response in Parkinson's disease. We propose two discrete hypotheses, 1) that scratching induced skin barrier damage or 2) neuroinflammation seen in Parkinson's disease are sufficient to autoinoculate collagen XVII in a tolerant with minimal T-cell tolerance. To test this hypothesis, we propose utilizing a novel animal model. We have used CRISPR-Cas9 to develop a mouse expressing a LoxP-PolyA-LoxP sequence upstream of exon 18, the coding region of the NC15a domain of collagen XVII. This domain corresponds with the human NC16a domain, which serves as the principle autoantigen in BP. When crossed to a tamoxifen inducible Cre-Lox mouse model, restoration of collagen XVII expression occurs in early adulthood, bypassing T-cell tolerance once unmasked. With inflammation and antigen unmasking through scratching or experimentally induced Parkinson's disease, we anticipate development of antigen-specific CD4+ T-cells and autoantibodies against collagen XVII. Two specific aims are proposed: 1) Determine the impact of scratching behaviors on the development of anti- collagen XVII autoantibodies, antigen-specific CD4+ T-cells, and the development of BP. 2) Determine the effect of neuroinflammation in experimental Parkinsonism in inducing cutaneous autoimmunity against collagen XVII. This project will provide significant insight into both the role of scratching and neuroinflammation in antigen unmasking. This proposal is likely to have strong translational potential across several disease processes and organ systems. Our novel approach provides a valuable tool for studying the immune response in Parkinson's disease, as well as for studying other autoimmune diseases with well-characterized autoantigens.