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

摘要：阿片类药物使用障碍患者的免疫图谱提示阿片类药物的结构导向设计。 特异性单抗 阿片类药物使用障碍(OUD)和阿片类药物相关过量是国家紧急状态。超过100,000 过量死亡发生在2020年4月至2021年4月期间，主要是由芬太尼引起的 单独或与其他阿片类药物或精神刺激剂混合。自新冠肺炎大流行爆发以来，有 需要住院治疗的非致命性过量用药有所增加。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- 特异性单抗是从抗原中分离出来的，或由疫苗诱导的多克隆抗体产生。 回应。与传统的以小分子为基础的处理方法相比 针对大脑MU阿片受体(MOR)的药物治疗，mAbs与药物结合并形成复合体 循环中的分子。因此，单抗的使用会阻碍药物通过血脑屏障。 通过隔离分子，削弱其中枢神经系统的作用。与MOR-配体相比，mAb提供了 更持久的疗效，不干扰非靶向药物。因此，这项拟议研究的重点是： 1)与分离配对的人阿片特异性B细胞受体(BCR)谱系的功能特征 2)结构导向的人源化和人源化单抗的设计，具有更好的疗效和 选择性。研究方法将涉及确定潜在单抗候选者的补充战略。 包括基于下一代测序的BCR测序、抗体展示和抗体工程，以 验证将OUD或疫苗配对导致的BCR基因变异性与结构导引的假设 抗体设计将识别出具有更大治疗潜力的单抗。此外，拟议的研究将告知 OUD和疫苗接种如何在BCR中引入抗原特异性遗传扰动。在了解了 抗体结构，本研究的结果将导致产生具有更高亲和力的单抗。要实现 这些目标，目标1描绘了人类bcr谱系在免疫OUD患者中的演变 一种羟考酮结合疫苗的I期临床试验(NCT04458545)和未免疫的OUD 病人。目的2检测阿片类药物特异性Fab结构与mAb效力和选择性的相关性。这个 这项研究的结果将扩大我们对人类阿片特异性抗体和B细胞谱系的了解， 支持基于结构的抗体工程，制备高亲和力的单抗。此外，结果还可以 加速发展以抗体为基础的策略，作为替代和补充的解决方案治疗 阿片类药物过量。

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.