Host Factors in Response to Therapeutic Monoclonal Antibodies and Vaccination

项目总结。Fc依赖的抗体介导的杀伤治疗消融的有效反应 需要宿主的细胞型特异性Fc受体与宿主的 治疗性单抗。许多患者缺乏有效的反应，可能高达30%， 这既是一个机会，也是一个挑战，以描绘这种差异的机制基础。 遗传研究可以确定宿主对这些差异的贡献。我们的初步数据 这表明，近三分之一的人在经典的Fc基因座上除了 影响配体结合亲和力、受体迁移率的单核苷酸多态现象 细胞膜和受体的定量表达。这些较大的结构变体影响Fc受体 在淋巴系和髓系细胞系中，近三分之一的变异体没有表现出 基因组结构、由此产生的蛋白质结构变化以及对净生物功能的影响。新的, 创新的技术方法，包括连锁阅读测序，现在与传统的聚合酶链式反应确认， 已经在基因组组织中展示了新的结构。CRISPR/CAS9靶向切除的应用 与连锁阅读测序相结合，现在能够解决新的结构变异 影响生物功能。再加上增强关键激活受体表达的可能性， 这是一个难得的机会，既可以增强宿主对治疗性单抗的反应，也可以增强 疫苗接种方案。因此，本提案的目标是1)通过我们的特征队列 具有不同祖先背景的捐赠者(N&GT；5000)，以表征小说的基因组组织 使用连锁阅读、位点特定切除和长阅读策略的结构变体；2)识别 预测新的受体蛋白及其结构，并评估其表达和功能，包括 可能的诱饵，信号缺陷的结构；以及3)开发一个可扩展的、可推广的平台来评估 人类FCGR基因座中SNPs和更大的基因组结构变异的研究 我们不断扩大的捐赠者池(10,000英镑)中的人口多样性，并评估预测对 治疗性抗体疗法。受体组合及其结构和表达的评估 将启用选择和分层接受者的战略，以实现最佳精准医学方法。

PROJECT SUMMARY. Effective responses in Fc-dependent, antibody-mediated killing for therapeutic ablation require efficient and productive interactions between the cell-type specific Fc receptors of the host and the therapeutic monoclonal antibody. The lack of effective responses in many patients, perhaps as much as 30%, represents both an opportunity and a challenge to delineate the mechanistic basis for such differences. Genetic inquiry can identify the contributions that the host brings to these differences. Our preliminary data indicate that nearly one-third of persons have structural variants (SV) in the classical Fc locus in addition to the prevalent single nucleotide polymorphisms affecting affinity of ligand binding, receptor mobility in the plane of the cell membrane and quantitative receptor expression. These larger structural variants affect Fc receptors on both lymphoid and myeloid cell series, and nearly a third of these variants are uncharacterized in terms of genomic structure, resultant alterations in protein structure and impact on net biological function. New, innovative technical approaches including linked-reads sequencing, now confirmed with conventional PCR, have demonstrated novel structures in genomic organization. Application of CRISPR/Cas9 targeted excision of the locus, coupled with linked-reads sequencing now enables resolution of novel structural variations impacting biological function. Coupled with the potential to enhance expression of key activating receptors, there is the exceptional opportunity to enhance both the host response to therapeutic mAbs but also to vaccination protocols. Accordingly, the aims of this proposal are 1) enabled by our characterized cohort of donors (N > 5,000) with different ancestral backgrounds, to characterize the genomic organization of novel structural variants using linked-read, locus specific excision and long read strategies; 2) to identify the predicted novel receptor proteins and their structures and assess their expression and function, including possible decoy, signaling deficient structures; and 3) to develop a scalable, generalizable platform to assess the repertoire of SNPs and larger genomic structural variants in the human FCGR genetic locus to understand the population diversity in our expanding donor pool (>10,000) and assess the ability to predict response to therapeutic antibody therapy. Assessment of the portfolio of receptors, their structures and their expression will enable strategies for selection and stratification of recipients for an optimal precision medicine approach.