ImmunoPET Probes for the Imaging of Lyme Disease

项目摘要/摘要 莱姆病是由伯氏疏螺旋体(BBSL)引起的一种扁虱传播的疾病，是 在美国最流行的壁虱传播疾病。疾控中心的最新监控报告称 莱姆病每年在美国影响超过47.5万人，比2015年增加了近50%。 此外，这种疾病正在从东北部、大西洋中部和上大西洋的流行区蔓延 中西部。与这种疾病相关的迅速增加的公共健康风险和不断上升的医疗费用是 围绕其诊断和治疗的争议加剧了这一问题。而莱姆的诊断不足 疾病导致治疗不足，疾病的误诊和误治已导致严重 发病率也很高。简而言之，我们目前对诊断、病理生理学和 缺乏莱姆病的治疗方法。综上所述，这些数据表明，开发用于 莱姆病的非侵入性分子成像是科学和临床的迫切需要。 这份R01提案的重点是合成、表征和体内验证一流的 用于莱姆病PET成像的放射性药物。这些药物的目标将是VlsE，一种蛋白质 在脊椎动物宿主BBSL的整个过程中，它的表面都有丰富的表达。具体目标1 (SA1)将集中于89Zr-的合成、化学表征和体外生物学评价。 标记的单抗和68Ga标记的单域抗体可与VlsE高度结合 专一性、选择性和亲和力。特定目标2(SA2)将集中在询问体内的 这些VlsE靶向放射性药物在两种莱姆病小鼠模型中的表现。首先，我们将 用皮下接种培养的BBSL的小鼠来询问其敏感性和特异性 并评估显像剂作为监测疗效的工具的能力。 抗生素治疗。随后，我们将在更多的时间内探索VlsE靶向PET探针的性能 高级莱姆病小鼠模型：小鼠通过肩部硬蜱感染多种BBSL菌株。在……里面 这两个模型，纵向PET数据将与一组体外分析技术一起使用，以评估 放射性示踪剂描绘活动性感染部位的能力。 最终，我们认为以BBSL为靶标的放射性示踪剂可能会对这两个领域产生范式转换的影响 莱姆病的基础科学与临床研究。在实验室，一种VlsE靶向放射免疫结合物 可作为研究BBSL的扩散、病理生理学和治疗的一种有价值的无创性工具 在疾病的小鼠模型中的感染。在临床上，BBSL靶向的放射性药物可能是一种 不可缺少的研究工具，以支持新疫苗、疗法和 以及一种有效的显像剂来监测患者的治疗反应 尤其是持续或严重的BBSL感染。

Project Summary/Abstract Lyme disease is a tick-borne disease caused by the spirochete Borrelia burgdorferi sensu lato (Bbsl) and is the most prevalent tick-borne disease in the United States. The latest surveillance from the CDC reported that Lyme disease affects more than ~475,000 people in the U.S. every year, a nearly 50% increase from 2015. Furthermore, the disease is expanding from its endemic areas in the Northeast, mid-Atlantic, and Upper Midwest. The rapidly increasing public health risks and rising healthcare costs associated with the disease are exacerbated by controversies surrounding its diagnosis and treatment. While the under-diagnosis of Lyme disease has led to under-treatment, the mis-diagnosis and mis-treatment of the condition have led to serious morbidity as well. Simply put, our current fundamental understanding of the diagnosis, pathophysiology, and treatment of Lyme disease is lacking. Taken together, the data indicate that the development of tools for the non-invasive molecular imaging of Lyme disease is an urgent scientific and clinical need. This R01 proposal is focused on the synthesis, characterization, and in vivo validation of first-in-class radiopharmaceuticals for the PET imaging of Lyme disease. The target for these agents will be VlsE, a protein that is abundantly expressed on the surface of Bbsl throughout its time in its vertebrate host. Specific Aim 1 (SA1) will be focused on the synthesis, chemical characterization, and in vitro biological evaluation of 89Zr- labeled monoclonal antibodies and 68Ga-labeled single domain antibodies capable of binding VlsE with high specificity, selectivity, and affinity. Specific Aim 2 (SA2) will be centered on interrogating the in vivo performance of these VlsE-targeted radiopharmaceuticals in two murine models of Lyme disease. First, we will use mice subcutaneously inoculated with cultured Bbsl to interrogate the sensitivity and specificity of the radioimmunoconjugates and to assess the power of the imaging agents as tools for monitoring response to antibiotic therapy. Subsequently, we will explore the performance of the VlsE-targeted PET probes in a more advanced murine model of Lyme disease: mice infected multiple strains of Bbsl via Ixodes scapularis ticks. In both models, longitudinal PET data will be used alongside a battery of ex vivo analytical techniques to evaluate the ability of the radiotracers to delineate sites of active infection. Ultimately, we contend that a Bbsl-targeting radiotracer could have a paradigm-shifting impact on both the basic science and clinical study of Lyme disease. In the laboratory, a VlsE-targeted radioimmunoconjugate could be a valuable non-invasive tool for studying the dissemination, pathophysiology, and treatment of Bbsl infections in murine models of disease. In the clinic, a Bbsl-targeted radiopharmaceutical could be an indispensable research tool to support the clinical development of novel vaccines, therapeutics, and diagnostics as well as an effective imaging agent to monitor the therapeutic response of patients with particularly persistent or severe Bbsl infections.