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Systematic, molecular level analysis of the Fc receptor ligation on antibody effector functions

Fc 受体连接对抗体效应子功能的系统分子水平分析

基本信息

批准号：
10533299
负责人：
GEORGE Georgiou GEORGIOU
金额：
$ 46.16万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-19 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10533299
关键词：
Affinity Anti-Infective Agents Antibodies Antigen-Antibody Complex Antigens Area Binding Biological Assay Biological Products Cell Cycle Kinetics Cell surface Cells Coupled Cytometry Data Development Effector Cell Engineering Event Fc Receptor Fc domain Glean Human IgG1 Immune checkpoint inhibitor Individual Kinetics Leukocytes Ligation Macrophage Malignant Neoplasms Maps Measurement Mediating Molecular Myelogenous Myeloid Cells Natural Killer Cells Outcome Pathogenicity Phagocytosis Phenotype Phosphopeptides Phosphorylated Peptide Play Process Property Role Signal Transduction Surface Synapses Testing Therapeutic Monoclonal Antibodies Therapeutic antibodies Work antibody-dependent cell cytotoxicity cell killing clinical development clinical efficacy crosslink cytokine cytotoxic cytotoxicity density immunological synapse formation improved infectious disease treatment inhibiting antibody microscopic imaging monocyte nano neutrophil novel pathogen phosphoproteomics receptor receptor binding receptor expression sound

项目摘要

PROJECT SUMMARY/ABSTRACT Fc-mediated antibody effector functions, primarily antibody dependent cell phagocytosis (ADCP) and antibody dependent cell cytotoxicity (ADCC), have been established to play a central role on the mechanism of action of therapeutic antibodies including anti-infective antibodies and immune checkpoint inhibitors. Effector functions are triggered by the crosslinking of Fc receptors (FcRs) expressed on cytotoxic leukocyte subsets following binding to target cells opsonized by multiple antibodies. Recent findings have highlighted the key role of myeloid- derived cells, on the clearance of opsonized pathogenic cells by ADCP/ADCC. Myeloid-derived effectors, express multiple Fc𝛾Rs, most relevant being the activating receptors Fc𝛾RI, Fc𝛾RIIa and Fc𝛾RIIIa and the inhibitory Fc𝛾RIIb. Because multivalent immune complexes (ICs) engage and activate all surface Fc𝛾Rs on myeloid-derived effectors to various degrees (depending on Fc𝛾R expression, Fc:FcR affinity, immune complex size and antigen density) the magnitude and kinetics of the ADCP and ADCC processes are determined by the integrated outcome of the activation of all surface FcRs to various degrees. The central hypothesis to be tested here is that the quantitative understanding of ADCP and ADCC by myeloid effectors triggered by the ligation of each individual FcR by taking advantage of bulk assays and high phenotypic content single-cell cytotoxicity assays together with phosphoproteomic data to map the specific signaling events on myeloid-derived effector cells, will be essential for providing a sound framework on how to engineer the Fc domain for optimal effector functions. This work will capitalize on our unique set of aglycosylated engineered Fc domains that bind with absolute selectivity and dialed-in affinity to each FcR type. In Specific Aim 1 we will exhaustively and quantitatively map the effector phenotypes (ADCC, ADCP, cytokine release, trogocytosis) performed by human macrophages and monocytes (as well as by neutrophils and by NK cells for thoroughness) triggered by ligation of each FcR as a function of affinity, IC target size and antigen density. In Sp. Aim 2 we will use novel high throughput single cell cytotoxicity assays and on-chip cytometry to determine the precise kinetics of immune synapse formation and cell killing in ADCC (or engulfment for ADCP) as a function of FcR expression levels on individual cells and to interrogate key relevant mechanistic aspects central to these processes. In Sp. Aim 3 we will use phosphoproteomics to: (a) identify and quantitate peptide phosphorylation events triggered by each FcR and (b) detect unique FcR ligation-induced phosphopeptide signatures that correlate with effector functions triggered by that receptor.

项目总结/文摘