IgE antibody responses to the oligosaccharide galactose-alpha-1,3-galactose (alpha-gal) in murine and human atherosclerosis

IgE 抗体对小鼠和人类动脉粥样硬化中寡糖半乳糖-α-1,3-半乳糖 (α-gal) 的反应

• 批准号: 10842540
• 负责人: Loren D Erickson
• 金额: $ 32.04万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10842540
• 关键词: Age; Allergens; Antibody Formation; Antibody Response; Antigens; Arterial Fatty Streak; Artificial Intelligence platform; Atherosclerosis; Automobile Driving; B-Lymphocytes; Benchmarking; Blood; CCR6 gene; CXCR4 gene; Cells; Classification; Clinical; Clinical Data; Communities; Computer software; Coronary Arteriosclerosis; Coronary artery; Custom; Cytometry; Data; Data Set; Dendritic Cells; Development; Disease; Disease Outcome; Frequencies; Galactose; Goals; Histology; Human; IgE; Immune; Immunoglobulin Class Switching; Immunoglobulin Switch Recombination; Instruction; Interleukin-4; Label; Linear Regressions; Link; Machine Learning; Measures; Mediating; Memory B-Lymphocyte; Modeling; Molecular; Multivariate Analysis; Mus; NF-kappa B; Neural Network Simulation; Noise; Oligosaccharides; Outcome; Patients; Performance; Peripheral Blood Mononuclear Cell; Phenotype; Phosphorylation; Phosphotransferases; Play; Production; Proteins; Regulation; Reporting; Research; Risk; Role; Running; STAT1 gene; STAT6 gene; Sampling; Serum; Severity of illness; Signal Transduction; Signaling Protein; Smoking Status; T-Lymphocyte; Testing; Training; Work; cell type; clinically relevant; cohort; deep neural network; design; file format; galactosyl-(1-3)galactose; high dimensionality; machine learning framework; monocyte; mouse model; novel; p38 Mitogen Activated Protein Kinase; parent grant; precision medicine; protein expression; sex; single-cell RNA sequencing; statistics; transcription factor; transcriptomics; ultrasound; virtual

Project Summary Increased total serum IgE levels are associated with coronary artery disease (CAD). However, the causal role of antigen-specific IgE in CAD remains largely unexplored. Recent work from our group and others provide evidence that humans with IgE sensitization to the mammalian oligosaccharide allergen a-gal have larger coronary artery plaques and unstable plaque features signifying increased CAD compared to those without IgE to a-gal. Despite these compelling human associative findings, no study to date has investigated the role of antigen-specific IgE as a driver of CAD severity and the molecular and cellular mechanisms mediating IgE sensitization to a-gal linked to atherosclerosis. We recently reported that humans with IgE sensitization to a-gal had a higher frequency of CCR6+ switched memory (SWM) B cells. Notably, consistent with the association of the IgE sensitization to a-gal and CAD, the amount of CCR6 on SWM B cells was associated with CAD severity. Transcriptomic analysis demonstrated that CCR6+ SWM B cells expressed higher IL-4R and STAT6 in subjects that were IgE a-gal+ compared to IgE a-gal-. Interestingly, IL-4 and STAT6 are important for B cell class switch recombination to IgE, suggesting that cells that make IL-4 may be important in IgE to a-gal production. Preliminary data using a novel mouse model deficient in NKT cells that are early producers of IL-4 show reduced levels of IgE to a-gal and implicates invariant NKT cells in the regulation of IgE antibody production to a-gal. Based on these human and murine data, the overarching objective of the parent grant is to investigate whether these factors and cells play a causal role in atherosclerosis development due to IgE sensitization to a-gal. A major component of these studies is to analyze single cell mass cytometry data derived from PBMCs of a second, independent and larger cohort of humans with CAD sensitized to a-gal allowing for more robust multivariate analysis and deeper interrogation of immune cell phenotypes that mark those at greatest risk. The overall goal of this research supplement is to make the mass cytometry data AI-ready with associated datasheets that contain the cell subtype annotations, protein type and activation state markers, cohort statistics, CAD severity measures and other relevant clinical variables (e.g., age, sex, smoking status, etc.). We will prepare noise filtered, normalized, batch corrected, cell type annotated CAD patient single cell mass cytometry data for AI applications and use an explainable machine learning framework to predict measures of CAD severity and identify cell types driving the predictions. AI-ready data will be shared and serve as a model of how to prepare patient single cell data to be AI-ready for precision medicine and other applications.

项目概要 血清总 IgE 水平升高与冠状动脉疾病 (CAD) 相关。然而，因果作用 CAD 中抗原特异性 IgE 的作用在很大程度上仍未被探索。我们小组和其他人最近的工作提供了 有证据表明，对哺乳动物寡糖过敏原 a-gal 具有 IgE 敏感性的人类具有更大的 与无 IgE 的患者相比，冠状动脉斑块和不稳定斑块特征表明 CAD 增加 到 a-gal。尽管有这些令人信服的人类联想发现，但迄今为止还没有研究调查过 抗原特异性 IgE 作为 CAD 严重程度的驱动因素以及介导 IgE 的分子和细胞机制 对与动脉粥样硬化有关的a-gal 过敏。我们最近报道称，具有 IgE 对 a-gal 过敏的人类 CCR6+ 开关记忆 (SWM) B 细胞的频率较高。值得注意的是，与协会一致 IgE 对 a-gal 和 CAD 的敏感性，SWM B 细胞上 CCR6 的量与 CAD 严重程度相关。 转录组分析表明，CCR6+ SWM B 细胞在受试者中表达更高的 IL-4R 和 STAT6 与 IgE a-gal- 相比，IgE a-gal+ 为 IgE a-gal+。有趣的是，IL-4 和 STAT6 对于 B 细胞类别转换很重要 重组为 IgE，表明产生 IL-4 的细胞可能在 IgE 到 a-gal 的产生中发挥重要作用。 使用缺乏 NKT 细胞（IL-4 早期产生者）的新型小鼠模型的初步数据显示，NKT 细胞的减少 a-gal 的 IgE 水平，并暗示恒定的 NKT 细胞参与调节 a-gal 的 IgE 抗体生产。 根据这些人类和小鼠数据，家长资助的首要目标是调查是否 由于 IgE 对 a-gal 敏感，这些因素和细胞在动脉粥样硬化的发展中起着因果作用。一个 这些研究的主要组成部分是分析来自第二个、 独立且更大的 CAD 人群对 a-gal 敏感，允许更稳健的多变量 对标记风险最大的免疫细胞表型进行分析和更深入的询问。总体目标 本研究补充的目的是使质谱流式细胞仪数据与包含相关数据表的 AI 做好准备 细胞亚型注释、蛋白质类型和激活状态标记、队列统计、CAD 严重程度测量 以及其他相关的临床变量（例如年龄、性别、吸烟状况等）。我们将准备过滤噪音， 用于 AI 应用的标准化、批量校正、细胞类型注释的 CAD 患者单细胞质谱流式细胞术数据 并使用可解释的机器学习框架来预测 CAD 严重程度并识别细胞类型 推动预测。 AI 就绪数据将被共享，并作为如何准备患者单细胞的模型 数据为人工智能做好了精准医疗和其他应用的准备。