Immunoprofiling of Opioid Use Disorder Patients to inform structure-guided design of opioid-specific monoclonal antibodies

阿片类药物使用障碍患者的免疫分析可为阿片类药物特异性单克隆抗体的结构指导设计提供信息

• 批准号: 10751233
• 负责人: Yue Zhang
• 金额: $ 3.5万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751233
• 关键词: Acceleration; Active Immunization; Address; Affinity; Antibodies; Antibody Affinity; Antibody Response; Antigens; B cell repertoire; B-Cell Antigen Receptor; B-Lymphocyte Subsets; B-Lymphocytes; B-cell receptor repertoire sequencing; Binding; Biomedical Research; Brain; COVID-19 pandemic; Circulation; Complex; Conjugate Vaccines; Data; Databases; Development; Diagnosis; Dose; Drug Targeting; Drug usage; Emergency Situation; Enrollment; Environment; Evolution; Fentanyl; Flow Cytometry; Frequencies; Future; Generations; Genetic; Goals; Half-Life; Heroin; Hospitalization; Human; Immunize; Immunoglobulin Somatic Hypermutation; In Vitro; Individual; Knowledge; Laboratories; Lead; Libraries; Ligands; Lymphocyte; Mature B-Lymphocyte; Memory B-Lymphocyte; Methods; Monoclonal Antibodies; Morphine; Mus; Mutate; Mutation; Naloxone; Opioid; Opioid Antagonist; Overdose; Oxycodone; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacotherapy; Phase I Clinical Trials; Phylogenetic Analysis; Population; Research; Research Project Grants; Roentgen Rays; Sampling; Selection Bias; Series; Signal Pathway; Site-Directed Mutagenesis; Sorting; Structure; Testing; Therapeutic; Translations; Vaccinated; Vaccination; Vaccines; Yeasts; analog; antibody engineering; antigen binding; blood-brain barrier crossing; cDNA Library; career; conventional therapy; design; fentanyl overdose; human monoclonal antibodies; human subject; humanized monoclonal antibodies; improved; lead series; mu opioid receptors; murine monoclonal antibody; next generation sequencing; novel therapeutics; opioid overdose; opioid use; opioid use disorder; overdose death; pharmacologic; polyclonal antibody; prevent; prophylactic; psychostimulant; response; single-cell RNA sequencing; skills; small molecule; statistics; synthetic opioid; unvaccinated; vaccine-induced antibodies

ABSTRACT: Immunoprofiling of Opioid Use Disorder Patients to inform structure-guided design of opioid- specific monoclonal antibodies The opioid use disorder (OUD) and opioid-related overdoses are a national emergency. Over >100,000 overdose deaths occurred in the period between April 2020 and April 2021, which are largely driven by fentanyl alone or mixed with other opioids or psychostimulants. Since the initiation of the COVID-19 pandemic, there has been an increase in non-fatal overdoses requiring hospitalization. These statistics clearly indicate that approved pharmacotherapies are not sufficient in preventing or treating OUD and opioid overdose. Antigen- specific monoclonal antibodies (mAbs) are isolated from the antigen, or by vaccine-induced polyclonal antibody response. Compared to conventional treatment methods based upon small molecule-based pharmacotherapies targeting the brain mu opioid receptor (MOR), mAbs bind and form a complex with the drug molecule in circulation. Hence, administration of mAbs impedes drugs from crossing blood-brain-barrier through sequestration of the molecules, blunting their CNS effects. Compared to MOR-ligands, mAb offer longer lasting efficacy and no interference with off-target drugs. Therefore, the proposing study focuses on: 1) functional characterization of the human opioid-specific B cell receptor (BCR) repertoire paired with isolation of opioid-specific mAb; 2) structure-guided design of humanized and human mAbs with greater efficacy and selectivity. The research approach will involve complementary strategies to identify potential mAb candidates including next-generation sequencing based-BCR sequencing, antibody display and antibody engineering, to validate the hypothesis that pairing OUD- or vaccine-induced BCR genetic variability with structure-guided antibody design will identify mAb with greater therapeutic potential. Moreover, the proposed study will inform us of how OUD and vaccination introduces antigen-specific genetic perturbations in BCRs. With knowledge of antibody structure, the result of this study will lead to generation of mAbs with improved affinity. To achieve these goals, AIM 1 delineates the evolution of human BCR repertoire in OUD patients who are immunized with a conjugated oxycodone-specific vaccine in Phase I clinical trials (NCT04458545) and unimmunized OUD patients. AIM 2 tests the relevance of the opioid-specific Fab structure to the mAb efficacy and selectivity. The results of the study will expand our understanding on the human opioid-specific antibody and B cell repertoire, supporting structural-based antibody engineering to generate mAbs with high affinity. Moreover, the results can accelerate the development of antibody-based strategy as an alternative and complementary solution treating opioid overdose.

摘要：阿片类药物使用障碍患者的免疫分析为阿片类药物的结构指导设计提供信息 特异性单克隆抗体 阿片类药物使用障碍（OUD）和与阿片类药物相关的过量用药是国家紧急状态。超过 >100,000 2020年4月至2021年4月期间发生的服药过量死亡主要是由芬太尼造成的 单独使用或与其他阿片类药物或精神兴奋剂混合使用。自 COVID-19 大流行开始以来， 需要住院治疗的非致命性用药过量的情况有所增加。这些统计数据清楚地表明 已批准的药物疗法不足以预防或治疗 OUD 和阿片类药物过量。抗原- 特异性单克隆抗体（mAb）是从抗原中分离出来的，或者是通过疫苗诱导的多克隆抗体 回复。与基于小分子的传统治疗方法相比 针对脑 mu 阿片受体 (MOR) 的药物疗法，单克隆抗体与药物结合并形成复合物 分子在循环中。因此，单克隆抗体的施用会阻止药物穿过血脑屏障 通过隔离分子，削弱它们的中枢神经系统作用。与 MOR 配体相比，mAb 提供 疗效更持久，且不干扰脱靶药物。因此，拟议的研究重点是： 1) 人类阿片类药物特异性 B 细胞受体 (BCR) 库的功能表征与分离 阿片类药物特异性单克隆抗体； 2）以结构为指导的人源化和人单克隆抗体设计，具有更高的功效和 选择性。研究方法将涉及补充策略来识别潜在的单克隆抗体候选者 包括基于下一代测序的BCR测序、抗体展示和抗体工程， 验证将 OUD 或疫苗诱导的 BCR 遗传变异与结构引导配对的假设 抗体设计将鉴定出具有更大治疗潜力的单克隆抗体。此外，拟议的研究将告知 我们了解 OUD 和疫苗接种如何在 BCR 中引入抗原特异性遗传扰动。凭借知识 抗体结构，这项研究的结果将导致产生具有改善亲和力的单克隆抗体。达到 为了实现这些目标，AIM 1 描述了接种了 OUD 疫苗的 OUD 患者中人类 BCR 库的演变 处于 I 期临床试验的结合羟考酮特异性疫苗 (NCT04458545) 和未免疫的 OUD 患者。 AIM 2 测试阿片类药物特异性 Fab 结构与 mAb 功效和选择性的相关性。这 研究结果将扩大我们对人类阿片特异性抗体和 B 细胞库的理解， 支持基于结构的抗体工程以生成具有高亲和力的单克隆抗体。此外，结果可以 加速基于抗体的策略的开发，作为治疗的替代和补充解决方案 阿片类药物过量。