Host Factors in Response to Therapeutic Monoclonal Antibodies and Vaccination

项目摘要。Fc依赖性抗体介导的抗病毒宿主防御中的有效应答 需要宿主的细胞类型特异性Fc受体与抗- SARS-CoV-2抗体。一些患者明显缺乏有效的反应， 这是一个机会，也是一个关键的挑战，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。初步数据，在我们的父母 U 01，表明近三分之一的人在经典Fc基因座中具有结构变异（SV）， 普遍的单核苷酸多态性影响配体结合的亲和力， 细胞膜平面和定量受体表达。这些较大的SV影响Fc受体， 淋巴细胞和骨髓细胞系列，近三分之一的基因组结构没有特征， 由此导致的蛋白质结构的改变和对净生物功能的影响。此外，非洲人- 在这些新的变异群中，美国遗产的代表性过高。目前的基因分型和 全基因组测序工作（例如，COVID-19 hg; COVIDhge）无法识别和定义这些SV 因为FCGR基因座的片段重复与旁系同源物具有>98%的同源性。新的， 创新的技术方法，涉及CRISPR/Cas9靶向切除旁系同源物，再加上长- 读段测序现在能够解析影响生物功能的新结构变异。等 知识将为我们提供难得的机会，以加强我们对COVID-19差异的理解 疾病，预测宿主反应，并可能调整疫苗接种方案。因此，目标 1）立即实施和应用我们的CRISPR/Cas9消化策略， Aim 1b的读取序列技术可以扩大最初提出的个人数量，强调 受COVID-19大流行影响的少数民族，2）扩大变异的数量 聚类以包括在非洲裔美国人供体中发现的所有那些，3）对100名患者的FCGR区域进行测序 严重的COVID-19表型，从整个南方腹地提取的生物标本中提取，以及4） 加速测序数据的分析，以确定新的结构，并开始开发 用于检测预测影响Fc受体的SV和SNP的可扩展基因分型策略 功能/表达，并影响急性和慢性炎症中宿主对获得性体液免疫的反应。 疾病的恢复期。这些目标将使我们的COVID-19企业生物库和 通过我们的CTSA驱动的CCTS合作伙伴网络， 影响了我们鉴定个体的能力，这些个体或多或少可能对效应抗体产生应答。

PROJECT SUMMARY. Effective responses in Fc-dependent, antibody-mediated anti-viral host defense require efficient and productive interactions between the cell-type specific Fc receptors of the host and anti- SARS-CoV-2 antibody. The apparent lack of effective responses in some patients represents both an opportunity and a critical challenge to delineate the mechanistic basis for such differences. Genetic inquiry can identify the contributions that the host brings to these differences. Preliminary data, presented in our parent U01, 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 SVs affect Fc receptors on both lymphoid and myeloid cell series, and nearly a third are uncharacterized in terms of genomic structure, resultant alterations in protein structure and impact on net biological function. Furthermore, persons of African- American heritage are over-represented in a number of these novel variant clusters. Current genotyping and whole genome sequencing efforts (eg, COVID-19 hg; COVIDhge) are unable to identify and define these SVs because of the segmental duplication of the FCGR locus with >98% homology of the paralogs. New, innovative technical approaches involving CRISPR/Cas9 targeted excision of the paralogs, coupled with long- read sequencing, now enable resolution of novel structural variations impacting biological function. Such knowledge will present the exceptional opportunity to enhance our understanding of disparities in COVID-19 disease, to anticipate the host response and potentially to adjust vaccination protocols. Accordingly, the goals of this supplement are to 1) immediately implement and apply our CRISPR/Cas9 digestion strategy and long- read sequence technology of Aim 1b to an expanded number of individuals originally proposed emphasizing ethnic minorities disproportionally affected by the COVID-19 pandemic, 2) expand the number of variant clusters to include all those found in African-American donors, 3) sequence the FCGR region in 100 patients with severe COVID-19 phenotype drawing on biospecimens drawn from across the Deep South, and 4) accelerate the analysis of sequencing data to identify the novel architectures and begin development of scalable genotyping strategies for detection of SVs and SNPs that are predicted to impact Fc receptor function/expression and influence the host response to acquired humoral immunity in both the acute and convalescent phases of disease. These goals will be enabled our COVID-19 Enterprise biobank and biospecimens from across the Deep South through our CTSA-driven CCTS Partner Network and will directly impact on our ability to identify individuals more or less likely to respond to effector antibody production.