Genetic basis for persistence of Borrelia burgdorferi

伯氏疏螺旋体持续存在的遗传基础

• 批准号: 10737678
• 负责人: Rafal Tokarz
• 金额: $ 55.98万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10737678
• 关键词: Address; Animal Model; Animals; Antibiotic Therapy; Antibody Response; Antigens; B-Lymphocytes; Biological Assay; Biology; Borrelia burgdorferi; Carditis; Catalogs; Cerebrospinal Fluid; Cognition Disorders; Coupled; DNA; Data; Detection; Development; Disease; Epitope Mapping; Epitopes; Exposure to; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Genome; Genomics; Genotype; Human; Immune response; Immunoglobulin G; Immunoglobulin M; Inbred BALB C Mice; Infection; Investigation; Link; Lyme Arthritis; Lyme Disease; Molecular; Molecular Analysis; Mus; Order Spirochaetales; OspC protein; Pathogenicity; Patients; Peptides; Peromyscus; Plasmids; Play; Post Treatment Lyme Disease Syndrome; Predisposing Factor; RNA; Role; Serology; Serotyping; Serum; Severity of illness; Specimen; Syndrome; Testing; Tick-Borne Diseases; Tissue Sample; Tissues; Variant; Vertebrates; Work; chronic infection; co-infection; cohort; density; detection test; experience; experimental study; genetic architecture; immunoreactivity; improved; insight; next generation; next generation sequencing; novel; pathogen; patient response; persistent symptom; prevent; response; specific biomarkers; tick-borne; tool; transcriptome

ABSTRACT The genetic factors required to establish a persistent infection in vertebrate hosts is one of the least understood aspects of an infection with Borrelia burgdorferi. Multiple triggers for establishment of persistence have been proposed. However, the lack of sensitive serologic and molecular assays has substantially impeded work examining the biology of persistent spirochetes and their direct role in Post-Treatment Lyme Disease Syndrome (PTLDS). We have developed two assays that overcome these major limitations and can facilitate studies of persistent infections. The TBDCapSeq, a next generation capture sequencing assay, has a sensitivity superior to quantitative PCR and enables B. burgdorferi genome assembly in host tissue samples. The TBD-Serochip, a peptide array that can identify antibody responses to 170,000 linear peptides per assay, provides unprecedented glimpses into B cell responses to the major antigens of B. burgdorferi. In this proposal these assays will be employed in three Specific Aims that will examine molecular and genetic footprints of a persistent B. burgdorferi infection. In Specific Aim 1, we will perform a comprehensive serologic analysis of sera and cerebrospinal fluid from a well characterized PTLDS patient cohort. We will identify all major immunoreactive epitopes in both specimen types. These data will be differentiated with serology data from other non-PTLDS Lyme disease cohorts to implicate potential specific biomarkers of PTLDS. In Specific Aim 2, we will employ the TBDCapSeq to identify molecular footprints of B. burgdorferi in the PTLDS patient cohort. The combination of superior sensitivity of TBDCapSeq coupled with high sequencing depth will provide the most robust molecular analysis of the link between B. burgdorferi and PTLDS. In Specific Aim 3, we will establish an animal model for three distinct genotypes of B. burgdorferi in C3H/HeN, BALB/c and Peromyscus leucopus mice. The capacity of each B. burgdorferi genotype for establishing persistence with and without antibiotic treatment will be assessed and the TBDCapSeq will be used to identify genetic features unique to persistent spirochetes. Along with genomic analysis, we will perform the first transcriptome investigation of persistent spirochetes. Combined, these Aims will provide insight into the immune response of patients with PTLDS and identify mechanistic triggers of spirochete persistence.

抽象的 在脊椎动物宿主中建立持续感染所需的遗传因素是最小的之一 理解了伯里亚·伯格多菲利（Borrelia Burgdorferi）感染的各个方面。建立持久性的多个触发器 已经提出了。但是，缺乏敏感的血清学和分子测定已极大地阻碍 研究持续螺旋体的生物学及其在治疗后莱姆病中的直接作用的工作 综合征（PTLD）。我们已经开发了两个克服这些主要限制的测定法，并可以促进 持续感染的研究。 TBDCAPSESQ是下一代捕获测序测定法，具有 敏感性优于定量PCR，并在宿主组织样品中启用B. burgdorferi基因组组装。 TBD-Serochip，一种肽阵列，可以识别每个测定法对170,000个线性肽的抗体反应， 为B细胞对B. burgdorferi的主要抗原的B细胞反应提供前所未有的瞥见。在此提案中 这些测定将用于三个特定目的，这些特定目的将检查A的分子和遗传足迹 持续的B. burgdorferi感染。在特定目标1中，我们将对 来自PTLDS患者队列的血清和脑脊液。我们将确定所有专业 两种样品类型的免疫反应性表位。这些数据将与血清学数据不同 其他非PTLDS莱姆病同志牵涉到PTLD的潜在特定生物标志物。在特定的目标2中， 我们将利用TBDCAPSEQ确定B. burgdorferi在PTLDS患者队列中的分子足迹。 TBDCAPSESQ与高测序深度相结合的优质敏感性的结合将提供 B. burgdorferi和PTLD之间的联系最强大的分子分析。在特定的目标3中，我们将 为C3H/HEN，BALB/C和PEROMYSCUS建立三种不同基因型的动物模型 白肉小鼠。每个B. burgdorferi基因型的能力在有和没有的情况下建立持久性 将评估抗生素治疗，并将使用TBDCAPSESQ来识别遗传特征 持续的螺旋体。除了基因组分析外，我们将进行首次对 持续的螺旋体。这些目标结合在一起，将为患者的免疫反应提供见解 PTLD并识别螺旋体持久性的机械触发器。