Seronegative Autoimmune Encephalopathies: Biomarker Discovery, Validation & Deep Phenotyping

血清阴性自身免疫性脑病：生物标志物的发现、验证

基本信息

批准号：
10558599
负责人：
Andrew McKeon
金额：
$ 39.75万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10558599
关键词：
Acceleration Address Age Antibodies Antigens Ataxia Autoimmune Autoimmune Diseases of the Nervous System Autoimmune encephalitis Axon Bacteriophages Biological Assay Biological Markers Brain Diseases Cells Cellular Assay Cerebellar Ataxia Characteristics Clinic Clinical Clinical/Radiologic Complement Complementary DNA Data Degenerative Disorder Diagnosis Disease Early Diagnosis Early treatment Encephalitis Encephalopathies Ethnic Origin Future Goals Health Hippocampus Human IL8 gene Immune Immune system Immunofluorescence Immunologic Immunoglobulin G Immunologics Immunoprecipitation Incidence Indirect Fluorescent Antibody Technique Indirect Immunofluorescence Infectious Encephalitis Investigation Laboratories Length Libraries Mass Spectrum Analysis Methods Mission Molecular Movement Disorders Mus Neurologic Neurons Nuclear Ocular Motility Disorders Outcome Pathogenicity Patients Peptides Phage ImmunoPrecipitation Sequencing Phenotype Prediction of Response to Therapy Process Protein Microchips Proteins Proteome Public Health Publishing Radiology Specialty Recombinant Proteins Research Research Personnel Serum Synapses Techniques Testing Therapeutic Trials Time Tissues Transfection Translations Undifferentiated United States National Institutes of Health Validation Western Blotting Work amphiphysin biomarker discovery biomarker identification biomarker validation chemokine clinical phenotype clinical practice cohort cytokine disability disease mechanisms study experience innovation neural neuroimmunology novel novel marker novel therapeutics post-COVID-19 screening sex specific biomarkers targeted treatment therapeutic development trial design

项目摘要

PROJECT SUMMARY Incidence of autoimmune encephalitis exceeds that of infectious cause, but 50% percent (half of approximate 4000/year in the US) are IgG biomarker negative. Seronegativity hinders diagnosis and pathophysiologic understanding in autoimmune encephalopathies (encephalitis, cerebellar ataxias and other movement disorders). Novel IgG characterization, which attempts to address this gap, is linear and slow occurring at a rate of 1-2 year (using standard tissue-based immunofluorescence assay [IFA] followed by western blots, immunoprecipitation, and mass spectrometry). The long-term goal is to molecularly, clinically and mechanistically characterize autoimmune encephalopathies, and develop targeted autoimmune therapies. The overall objectives in this application, are to: 1) identify disease-specific biomarkers for seronegative autoimmune encephalopathies; 2) molecularly validate novel biomarkers, and 3) clinically, radiologically and immunologically deep-phenotype autoimmune encephalopathies. The central hypothesis is that seronegative autoimmune encephalopathies are characterizable though biomarker discovery. The rationale for this project is that biomarker discovery and translation could occur expeditiously, by leveraging complementary techniques in parallel, that novel biomarkers could be molecularly validated, and that disorders associated with new biomarkers could be neurologically and immunologically deep-phenotyped. To test the central hypothesis the following three specific aims will be pursued: 1) Identify novel biomarkers, initially in 3 partly characterized seronegative autoimmune encephalopathies; 2) determine validity of novel biomarkers; and 3) assess for differentiated clinical phenotypes, and cytokine profile accompaniments. Under the first aim, serum and CSF from seronegative autoimmune encephalopathy patients will be interrogated for neural antibodies using native full-length protein microarrays, as principle technique, complemented by tissue and cell-based IFA for initial IgG screening, and, as needed, phage immunoprecipitation sequencing, and mass spectrometry techniques for verification. For the second aim, newly characterized IgGs will be validated in autoimmune encephalopathy patients and controls by multiple antigen-specific methods (confocal indirect immunofluorescence, and recombinant protein assays [western blot, cDNA-transfected cell-based]). For the third aim, patients with autoimmune encephalopathies with characterized and validated neural IgGs will be deep-phenotyped clinically, radiologically and immunologically, by evaluating for IgG effects in live neuron assays, and cytokine-chemokine profiles. Remaining seronegative cohorts will be immunologically profiled by cytokine-chemokine assays. The research proposed is innovative in the applicant’s opinion, as it focuses on multiplexed biomarker identification, validation, and deep phenotyping. The proposed research is significant because disease-specific biomarkers, with in-depth characterization, allow earlier diagnosis and treatment, and support disease mechanism studies.

项目概要自身免疫性脑炎的发病率高于感染性脑炎，但约占 50%（大约一半）在美国每年有 4000 例）IgG 生物标志物呈阴性，这会阻碍诊断和病理生理学。了解自身免疫性脑病（脑炎、小脑性共济失调和其他运动障碍）试图解决这一差距的新型 IgG 表征是线性的且缓慢发生。 1-2 年的比率（使用标准的基于组织的免疫荧光 [IFA]，然后进行蛋白质印迹分析，免疫沉淀和质谱）的长期目标是分子、临床和质谱。从机制上表征自身免疫性脑病，并开发有针对性的自身免疫疗法。本申请的总体目标是：1) 识别血清阴性的疾病特异性生物标志物自身免疫性脑病；2) 分子验证的新型生物标志物，以及 3) 临床、放射学和免疫学深表型自身免疫性脑病的中心假设是血清阴性。自身免疫性脑病可以通过生物标志物的发现来表征。通过利用互补技术，生物标志物的发现和翻译可以迅速发生与此同时，新的生物标志物可以通过分子验证，并且与新的生物标志物相关的疾病生物标志物可以在神经学和免疫学上进行深层表型测试以检验中心假设。将追求以下三个具体目标： 1) 识别新的生物标志物，最初是 3 个部分特征化的生物标志物血清阴性自身免疫性脑病；2) 确定新型生物标志物的有效性；3) 评估不同的临床表型和细胞因子谱伴随在第一个目标下，血清和脑脊液。血清阴性自身免疫性脑病患者将使用天然抗体进行神经抗体询问全长蛋白质微阵列作为主要技术，并辅以基于组织和细胞的 IFA 进行初始 IgG 筛选，并根据需要进行噬菌体免疫沉淀测序和质谱技术为了验证第二个目标，新表征的 IgG 将在自身免疫性脑病中得到验证。通过多种抗原特异性方法（共聚焦间接免疫荧光法和重组蛋白检测[蛋白质印迹，基于 cDNA 转染细胞])。具有特征和验证的神经 IgG 的自身免疫性脑病将在临床上进行深度表型分析，放射学和免疫学，通过评估活神经元测定中的 IgG 效应以及细胞因子-趋化因子剩余的血清阴性群体将通过细胞因子-趋化因子测定进行免疫学分析。申请人认为所提议的研究具有创新性，因为它侧重于多重生物标志物识别，验证和深度表型分析具有重要意义，因为疾病特异性生物标志物，具有深入的表征，可以实现早期诊断和治疗，并支持疾病机制研究。