Molecular and functional investigation of the role of HLA-F in immune regulation

HLA-F在免疫调节中作用的分子和功能研究

• 批准号: 10636894
• 负责人: Erin June Adams
• 金额: $ 66.7万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-06 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10636894
• 关键词: 3-Dimensional; Address; Adenoviruses; Adopted; Amino Acids; Antibodies; Antigens; Applications Grants; Area; Autoimmune Diseases of the Nervous System; Autoimmunity; Binding; Biological Assay; Biophysics; Cell surface; Cells; Clone Cells; Data; Decidua; Disease; Effector Cell; Environment; Family; Family member; Funding; Genetic Polymorphism; Goals; HIV; HIV Infections; Health; Homeostasis; Human; Immune; Immunity; Infection; Invaded; Investigation; KLRD1 gene; Length; Ligands; Link; Major Histocompatibility Complex; Malignant Neoplasms; Methods; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; Natural Killer Cells; Peptides; Play; Post-Translational Protein Processing; Pregnancy; Production; Protein Isoforms; Proteins; Publishing; RNA Splicing; Reagent; Receptor Cell; Recombinants; Reproduction; Role; Source; Specificity; Structure; T-Lymphocyte; Testing; Variant; Viral; Work; biophysical analysis; cancer cell; cell type; conformer; fetal; immunoregulation; member; pathogen; protein complex; receptor; three dimensional structure; trafficking; tumor

Project Summary/Abstract HLA-F is a nonclassical class I MHC (Ib) molecule that has been found expressed on a variety of cancers, shown to play a role in HIV and adenoviral infection, the neurological autoimmune disease ALS and is expressed throughout pregnancy. Despite the potential importance of this protein in these conditions, little is known about this molecule in terms of its function or even in which conformational state it is expressed. We have recently shown that, in addition to being expressed as a heavy chain only state, or open conformer (HLA-FOC), HLA-F can also be expressed as a bon fide peptide presenting molecule, associated with the β2m subunit (pHLA-F). Peptides are presented in an unconventional way, with the N-terminus not anchored within the groove and the potential for post-translational modifications featuring in peptide anchoring. Despite these advances, there remains much unknown about how these conformer states are regulated, how it engages its various receptors in each of these conformer states, and the role of HLA-F in its various environments of tumor surveillance, autoimmunity and reproduction. Thus, the aims of this proposal focus on addressing these questions and are: Aim 1: To investigate, structurally and functionally, the various conformer states that HLA-F adopts in human health and disease. We will pursue structural studies of the HLA-F isoforms to understand how these two states differ from each other. Using conformer-specific antibodies, we will determine what cell types express which (or both) forms and how this differs between healthy and disease cells. We will also pursue peptide elution studies from a range of human sources to determine if the peptide repertoire shifts depending on cellular origin or disease. Aim 2: To identify and analyze the factors that regulate the production or interchange of HLA- F conformers and splice forms in a cell. We will explore the cellular factors that may play a role in switching HLA-F between peptide-loaded and HLA-FOC as well as an intriguing splice variant of HLA-F of unknown function. Finally, in Aim 3 we seek to establish the receptor repertoire that engage HLA-F in its various conformer states, determine the molecular basis for their association and study the functional consequences of their binding. We will employ the structural, biophysical and functional expertise of the Adams lab to determine the receptor repertoire that engage these conformer states of HLA-F and study them at the functional and molecular level.

项目概要/摘要 HLA-F 是一种非经典 I 类 MHC (Ib) 分子，已发现在多种癌症中表达， 显示在 HIV 和腺病毒感染、神经性自身免疫性疾病 ALS 中发挥作用，并被表达 尽管这种蛋白质在这些情况下具有潜在的重要性，但人们对此知之甚少。 我们最近对这种分子的功能甚至其表达的构象状态进行了研究。 结果表明，除了表达为仅重链状态或开放构象异构体 (HLA-FOC) 外，HLA-F 也可以表达为与 β2m 亚基 (pHLA-F) 相关的真正的肽呈递分子。 肽以非常规方式呈现，N 末端未锚定在凹槽内，并且 尽管有这些进展，但以肽锚定为特征的翻译后修饰的潜力仍然存在。 关于这些构象异构体状态是如何调节的，以及它如何与各种受体结合，仍然知之甚少 在每种构象异构体状态中，以及 HLA-F 在其各种肿瘤监测环境中的作用， 因此，该提案的目标是解决这些问题： 目标 1：从结构和功能上研究 HLA-F 在 我们将进行 HLA-F 亚型的结构研究，以了解这些亚型是如何发挥作用的。 两种状态彼此不同，我们将使用构象异构体特异性抗体来确定表达哪些细胞类型。 哪种（或两种）形式以及健康细胞和疾病细胞之间的差异我们还将研究肽洗脱。 来自一系列人类来源的研究，以确定肽库是否根据细胞起源而变化 目标 2：识别并分析调节 HLA- 产生或交换的因素。 细胞中的 F 构象异构体和剪接形式 我们将探讨可能在转换中发挥作用的细胞因素。 肽负载和 HLA-FOC 之间的 HLA-F 以及功能未知的 HLA-F 的有趣剪接变体。 最后，在目标 3 中，我们寻求建立以各种构象异构体结合 HLA-F 的受体库 状态，确定其关联的分子基础并研究其功能后果 我们将利用亚当斯实验室的结构、生物物理和功能专业知识来确定它们的结合。 参与 HLA-F 构象异构体状态的受体库，并在功能和功能上研究它们 分子水平。