Modulation of immunodominance in HLA class I associated uveitides

摘要/项目摘要 ERAP1的变异调节了几种形式的非感染性葡萄膜炎的风险- 相关的人类白细胞抗原I类，强烈表明免疫优势的变化迄今未得到证实，其影响 疾病的起因和防护。 本应用的总体目标是确定ERAP1同种异型通过哪种机制引起和 预防人类白细胞抗原I相关的葡萄膜炎。我们的长期目标是了解并在治疗上针对人类白细胞抗原 我把自身免疫与葡萄膜炎联系在一起。 我们的中心假设是，异型ERAP1改变了与HLAI结合的多肽，使其包括 当由疾病相关的HLAI呈递时是免疫原性的，它通过一种 免疫优势的改变。这项研究的基本原理是对ERAP1介导的机制的理解 HLAI免疫的发病机制将使针对特定ERAP1-HLAI-葡萄膜炎的靶向治疗设计成为可能 子集。 基于这些考虑，我们将实现两个具体目标。在目标1中，我们将通过 同种异型ERAP1 Hap10在人类白细胞抗原B*51白塞氏葡萄膜炎(BU)中启动免疫功能障碍的机制 但通过一系列CRISPR/Cas9基因组编辑保护人类白细胞抗原B*27急性前葡萄膜炎(AAU) 实验。这些实验将使我们能够确定其对限制的人类白细胞抗原I的功能贡献 与这些疾病相关的多肽及其对免疫原性或耐受性产生的影响 免疫反应。在目标2中，我们将确定人类白细胞抗原I限制的致病表位如何依赖于同种异型- 活化的BU和AAU中克隆扩增的CD8 T细胞的特异性ERAP1功能 患者进行细胞克隆、基因组编辑和免疫原性功能评估。 我们期望以下结果：1)了解免疫原性和耐受性所介导的影响 通过两个高度相关的疾病相关的人类白细胞抗原-B27和B51中的同种异型ERAP1，2)识别 在这些情况下诱导或阻止免疫原性的表位，3)异型ERAP1的原理证明 调节自身免疫，以及对其活性的操纵调节致病性，提供无可辩驳的 靶向ERAP1酶活性的药理基础，或通过基因治疗。这将会有一个 通过识别分子靶点对该领域产生积极影响，从而使治疗方法的设计得以实现 由基因类型定义的患者群体，并通过超出 BU和AAU对其他对人类健康有巨大影响的MHC-I疾病，如IBD、牛皮癣和 银屑病关节炎。

ABSTRACT/PROJECT SUMMARY Variants at ERAP1 modulate the risk for several forms of non-infectious uveitis in the presence of disease- associated HLA class I, strongly suggesting a so far unproven change in immunodominance with impact on disease causation and protection. The overall objective of this application is to determine through which mechanism ERAP1 allotypes cause and protect from HLA I-associated uveitis. Our long-term goal is to understand and therapeutically target HLA class I associated autoimmunity in uveitis. Our central hypothesis is that allotypic ERAP1 alters the HLA I-bound peptidome to include epitopes that are immunogenic when presented by disease-relevant HLA I, which induces or controls disease through a change in immunodominance. The rationale for this study is that mechanistic understanding of ERAP1-mediated pathogenesis in HLA I immunity will enable targeted therapy design aimed at specific ERAP1-HLA I-uveitis subsets. Based on these considerations we will implement two specific aims. In Aim 1 we will determine through which mechanism the allotype ERAP1 Hap10 initiates immune dysfunction in HLA-B*51+ Behçet’s uveitis (BU) but protects from HLA-B*27+ acute anterior uveitis (AAU) via a series of CRISPR/Cas9 genome editing experiments. These experiments will allow us to define its functional contribution to the HLA I restricted peptidomes relevant to each of these disorders and their effect on the generation of immunogenic or tolerogenic immune responses. In Aim 2 we will establish how HLA I restricted pathogenic epitopes depend on allotype- specific ERAP1 function through the exploitation of clonally expanded CD8 T cells from active BU and AAU patients for cellular cloning, genome-editing and functional assessment of immunogenicity. We expect the following outcomes 1) knowledge of the immunogenic and tolerogenic effects mediated by allotypic ERAP1 in two highly disease-relevant HLA restriction contexts: HLA-B27 and B51, 2) identity of epitopes that induce or prevent immunogenicity in these contexts, 3) proof of principle that allotypic ERAP1 regulates autoimmunity and that manipulation of its activity modulates pathogenicity providing irrefutable rationale for targeting ERAP1 enzyme activity pharmacologically, or through gene therapy. This will have a positive impact on the field through the identification of molecular targets allowing the design of therapy for patient groups defined by genotypes, and through mechanistic understanding extending beyond the scope of BU and AAU to additional MHC-I-opathies with immense impact on human health, such as IBD, psoriasis, and psoriatic arthritis.