Host Factors in Response to Therapeutic Monoclonal Antibodies and Vaccination

项目摘要。治疗性消融的Fc依赖性、抗体介导杀伤的有效应答 需要宿主的细胞类型特异性Fc受体和宿主的细胞类型特异性Fc受体之间的有效和生产性相互作用。 治疗性单克隆抗体。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械基础。 基因研究可以确定宿主对这些差异的贡献。我们的初步数据 表明近三分之一的人在经典Fc基因座中除了具有结构变体（SV）外， 普遍存在的单核苷酸多态性影响配体结合的亲和力，受体在 细胞膜和定量受体表达。这些较大的结构变体影响Fc受体 在淋巴和骨髓细胞系列，近三分之一的这些变体是未表征的方面， 基因组结构，蛋白质结构的改变和对净生物功能的影响。新的， 创新的技术方法，包括连接读数测序，现在用传统的PCR证实， 已经展示了基因组组织的新结构。CRISPR/Cas9靶向切除的应用 基因座，再加上连接阅读测序，现在可以解决新的结构变异 影响生物功能。再加上增强关键活化受体表达的潜力， 存在增强宿主对治疗性单克隆抗体的应答以及增强宿主对治疗性单克隆抗体的应答的特殊机会， 疫苗接种方案。因此，本提案的目的是：1）通过我们的特征化队列， 不同祖先背景的供体（N> 5，000），以表征新的 使用连接读段、基因座特异性切除和长读段策略来鉴定结构变体; 2）鉴定结构变体， 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;和3）开发一个可扩展的，可推广的平台，以评估 人类FCGR基因座中SNP和较大基因组结构变异的库，以了解 我们不断扩大的供体库（> 10，000）中的人口多样性，并评估预测 治疗性抗体疗法评估受体组合、其结构及其表达 将使战略的选择和分层的收件人，以最佳的精确医学方法。

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.