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Molecular determinants driving diverse mechanisms of antibody-mediated pathology

驱动抗体介导病理学多种机制的分子决定因素

基本信息

批准号：
10454388
负责人：
Kevin C O'connor
金额：
$ 20.94万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-20 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10454388
关键词：
Acetylcholine Address Adopted Affect Antibodies Antigens Autoantibodies Autoimmune Autoimmune Diseases Automobile Driving B-Lymphocytes Binding Binding Sites Biological Assay Biological Response Modifier Therapy CD46 Antigen CRISPR/Cas technology Cell Line Cholinergic Receptors Classical Complement Pathway Clinical Cloning Collection Communication Complement Complement Activation Complement Inactivators Complex Complex Mixtures Consequentialism Detection Diagnosis Disease Disease Progression Dissection Epitopes Frequencies Genes Heterogeneity Human Impairment Individual Investigation Ion Channel Knock-out Knowledge Laboratories Libraries Link Mammalian Cell Measures Mediating Modeling Molecular Monitor Monoclonal Antibodies Muscle Muscle Weakness Myasthenia Gravis Nerve Neuromuscular Diseases Neuromuscular Junction Neuromuscular Junction Diseases Neuromyelitis Optica Nicotinic Receptors Pathogenicity Pathology Patients Persons Prediction of Response to Therapy Prevalence Production Property Public Health Recombinants Resources Series Serum Severities Severity of illness Somatic Mutation Specificity Technology Testing Tissues Treatment outcome crosslink experience experimental study human monoclonal antibodies neuromuscular neuromuscular transmission novel novel strategies novel therapeutics pathogenic autoantibodies personalized therapeutic prevent receptor response sample fixation tool treatment response variable region gene

项目摘要

Project Summary. Myasthenia gravis (MG) is an autoimmune disorder of neuromuscular transmission. The most common subtype is characterized by autoantibodies (autoAbs) targeting the acetylcholine receptor (AChR), a membrane channel expressed on the muscle end-plate at the neuromuscular junction. The pathology is directly due to the AChR autoAbs. The AChR autoAbs in the serum of patients are broadly heterogeneous in their specificity as it includes autoAbs that recognize each of the four different subunits (a, b, e or d) of AChR. Furthermore, AChR autoAbs can use three distinct mechanisms to effect pathology: (i) complement-directed tissue damage, (ii) blocking the binding site for acetylcholine and (iii) modulation (internalization) of the AChR. Although AChR autoAbs are pathogenic, titers vary widely between different patients and within individuals during the course of their disease; importantly, titer neither correlates well with clinical severity nor predicts treatment outcome. Furthermore, new biological therapeutic strategies that specifically target AChR autoAb complement activation or reduce circulating titers have shown wide variability in response including patients that do not respond. The disassociation between titer and disease severity, and the heterogeneous response to autoantibody targeting treatments are not well understood. This gap in our knowledge exists first because AChR autoAbs are often studied with patient serum, which are confounded by the complex mixture of specificities. Second, existing assays only measure AChR autoAb binding, but not their pathogenic capacity or mechanism. We will address these gaps in our knowledge by (i) generating rare human monoclonal AChR autoAbs, (ii) using novel approaches to measure the different mechanisms of autoAb pathogenicity, and (iii) associating the autoAb properties with pathogenic capacity. To this end, we applied an unbiased high-throughput approach for producing AChR-specific human mAbs by cloning B cells, which circumvented technical challenges associated with cloning mAbs targeting this multi-subunit membrane channel. This will afford production of an AChR specific mAb library, which includes mAbs that recognize different subunits and epitopes within those subunits. New approaches to study the different effector mechanisms of AChR autoAb pathogenicity have also been developed. We will measure AChR autoAb-mediated acetylcholine blocking and AChR modulating functions of the AChR mAbs. Importantly, an assay for measuring complement activation, in which the complement inhibitors (CD46, CD55 and CD59) are knocked out, will be used to test AChR autoAb-dependent complement activity. These experiments will define how the specificity and molecular properties of AChR autoAb are associated with pathogenic effector function. Overall, this investigation will (i) provide a set of well-characterized mAbs which will serve as tools for more accurate modeling of AChR autoAb pathology; (ii) provide a framework for understanding the association between epitope specificity and complement activation and (iii) identify potential new therapeutic avenues for treating MG through specifically targeting AChR autoAb pathogenic mechanisms.

项目摘要。重症肌无力（MG）是一种神经肌肉传递的自身免疫性疾病。的最常见的亚型的特征在于靶向乙酰胆碱受体（AChR）的自身抗体（autoAb），在神经肌肉接头处的肌肉终板上表达的膜通道。病理学直接是因为乙酰胆碱受体抗体患者血清中的AChR自身抗体在其自身免疫应答中具有广泛的异质性。特异性，因为它包括识别AChR的四个不同亚基（a、B、e或d）中的每一个的自身抗体。此外，AChR autoAb可以使用三种不同的机制来影响病理学：组织损伤，（ii）阻断乙酰胆碱的结合位点和（iii）AChR的调节（内化）。虽然AChR自身抗体是致病性的，但不同患者之间和个体内的滴度差异很大重要的是，滴度既不能很好地与临床严重程度相关，也不能预测治疗结果此外，特异性靶向AChR autoAb的新生物治疗策略补体激活或降低循环滴度的反应显示出广泛的变异性，不要回答滴度和疾病严重程度之间的分离，以及对自身抗体靶向治疗还没有被很好地理解。我们知识上的这种差距首先是因为AChR autoAb通常与患者血清一起研究，其被复杂的特异性混合物混淆。其次，现有的检测方法仅测量AChR autoAb结合，而不测量其致病能力或机制。我们将通过（i）产生罕见的人单克隆AChR autoAb，（ii）使用新的方法来测量自身抗体致病性的不同机制，以及（iii）将具有致病能力的自身抗体特性。为此，我们采用了一种无偏的高通量方法，通过克隆B细胞生产AChR特异性人单克隆抗体，这规避了相关的技术挑战，克隆针对这种多亚基膜通道的mAb。这将提供生产乙酰胆碱受体特异性 mAb文库，其包括识别不同亚基和这些亚基内的表位的mAb。新还开发了研究AChR自身抗体致病性的不同效应机制的方法。我们将测量AChR autoAb介导的乙酰胆碱阻断和AChR的调节功能，单克隆抗体重要的是，一种用于测量补体激活的测定，其中补体抑制剂（CD 46， CD 55和CD 59）被敲除，将用于测试AChR autoAb依赖性补体活性。这些实验将确定AChR自身抗体的特异性和分子特性如何与致病效应子功能总体而言，本研究将（i）提供一组表征良好的mAb，作为AChR autoAb病理学更准确建模的工具;（ii）提供理解的框架表位特异性和补体激活之间关联和（iii）鉴定潜在的新治疗剂通过特异性靶向AChR autoAb致病机制治疗MG的途径。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Novel pathophysiological insights in autoimmune myasthenia gravis.

DOI：
10.1097/wco.0000000000001088
发表时间：
2022-10-01
期刊：
Current opinion in neurology
影响因子：
4.8
作者：
通讯作者：

MOGAD patient autoantibodies induce complement, phagocytosis, and cellular cytotoxicity.

DOI：
10.1172/jci.insight.165373
发表时间：
2023-06-08
期刊：
JCI INSIGHT
影响因子：
8
作者：
Yandamuri, Soumya S.;Filipek, Beata;Obaid, Abeer H.;Lele, Nikhil;Thurman, Joshua M.;Makhani, Naila;Nowak, Richard J.;Guo, Yong;Lucchinetti, Claudia F.;Flanagan, Eoin P.;Longbrake, Erin E.;O'Connor, Kevin C.
通讯作者：
O'Connor, Kevin C.

The clinical need for clustered AChR cell-based assay testing of seronegative MG.

基于ACHR细胞的临床需求基于ACHR细胞的测定测试。

DOI：
10.1016/j.jneuroim.2022.577850
发表时间：
2022-06-15
期刊：
JOURNAL OF NEUROIMMUNOLOGY
影响因子：
3.3
作者：
Masi, Gianvito;Li, Yingkai;Karatz, Tabitha;Pham, Minh C.;Oxendine, Seneca R.;Nowak, Richard J.;Guptill, Jeffrey T.;'Connor, Kevin C. O.
通讯作者：
'Connor, Kevin C. O.

Remission of severe myasthenia gravis after autologous stem cell transplantation.

DOI：
10.1002/acn3.51898
发表时间：
2023-11
期刊：
ANNALS OF CLINICAL AND TRANSLATIONAL NEUROLOGY
影响因子：
5.3
作者：
Schlatter, Monica I.;Yandamuri, Soumya S.;O'Connor, Kevin C.;Nowak, Richard J.;Pham, Minh C.;Obaid, Abeer H.;Redman, Callee;Provost, Marie;Mcsweeney, Peter A.;Pearlman, Michael L.;Tees, Michael T.;Bowen, James D.;Nash, Richard A.;Georges, George E.
通讯作者：
Georges, George E.

Myasthenia gravis complement activity is independent of autoantibody titer and disease severity.