Anaplasma phagocytophilum modulate tick gene expression for its survival and transmission from the vector host

嗜吞噬细胞无形体调节蜱基因表达以使其存活并从载体宿主传播

• 批准号: 9398343
• 负责人: Girish Neelakanta
• 金额: $ 38.75万
• 依托单位: OLD DOMINION UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-18 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9398343
• 关键词: Actins; Address; Adult; Affect; Anaplasma phagocytophilum; Antibodies; Antifreeze; Area; Arthropod Vectors; Arthropods; Bacteria; Binding; Biochemical; Biological Assay; Biology; Black-legged Tick; Bovine Anaplasmosis; Cells; Confocal Microscopy; Data; Development; Disease; EMSA; Electrophoretic Mobility Shift Assay; Enzymes; Female; Gene Expression; Genes; Goals; Human; Immunization; In Vitro; Interruption; Kynurenine-oxoglutarate aminotransferase; Lead; Mammalian Cell; Mammals; Medical; Microbe; Modeling; Molecular; Molecular Analysis; Neuromodulator; OATP Transporters; Oxidative Stress; Pathway interactions; Phosphorylation; Promoter Regions; RNA Interference; Regulation; Reproduction; Research; Rickettsia; Rickettsiales; Role; Salivary Glands; Signal Pathway; Signal Transduction; Tick-Borne Diseases; Ticks; Tryptophan; Tryptophan Metabolism Pathway; United States; Vertebrates; Xanthurenic Acid; base; design; egg; experimental study; field study; in vivo; insight; knock-down; male; mouse model; neutrophil; novel; pathogen; transmission process; vector; vector transmission

Project Summary/Abstract Human anaplasmosis, caused by the obligate intracellular bacterium Anaplasma phagocytophilum, is one of the most common tick-borne diseases in the United States. In mammals and ticks, A. phagocytophilum resides in neutrophils and in salivary glands, respectively. Despite numerous studies that have focused on understanding strategies that A. phagocytophilum uses to survive in the mammalian cells, relatively few studies have clearly defined the molecular strategies that A. phagocytophilum uses to survive in ticks. In this project, we will be performing a comprehensive molecular analysis on Ixodes scapularis organic anion transporting polypeptides (OATPs) and genes involved in the tryptophan metabolism pathway in A. phagocytophilum-tick interactions. This study is build upon our efforts in showing how A. phagocytophilum modulates gene expression and cell signaling for its survival in the vector. As an example, our previous studies has shown that A. phagocytophilum modulate tick antifreeze gene expression and actin phosphorylation for its survival in the vector. Here, we provide strong preliminary in vitro and in vivo data showing that A. phagocytophilum induces expression of a specific OATP (OATP4056) and kynurenine aminotransferase (KAT), a gene involved in the tryptophan pathway. We now hypothesize that interplay between OATP4056 and KAT not only facilitates A. phagocytophilum survival in the vector but also aid in the transmission of this bacterium to a vertebrate host. RNAi analysis revealed that knockdown of OATP4056 has no effect on A. phagocytophilum acquisition but affected its survival and transmission from these ticks. Electrophoretic mobility shift assays with promoter region and total lysates prepared from uninfected and A. phagocytophilum-infected ticks provide further evidence that the presence of this bacterium influences expression of OATP4056 in these ticks. In addition, we found evidence that a metabolite from tryptophan pathway regulates A. phagocytophilum survival and OATP4056 gene expression in these ticks. These results provide important insights for our proposed studies to define molecular basis of the relationship of A. phagocytophilum with ticks. Based on our strong preliminary results and the experiments proposed, we believe that this could be a transformative study that not only serves as a model to study intimate relationships established by pathogens with their arthropod vectors but may also lead in the development of new strategies to interrupt the transmission of this and perhaps other Rickettsial species of medical importance.

项目摘要/摘要 人类无浆体病，由专性细胞内细菌吞噬无浆菌引起， 是美国最常见的扁虱传播疾病之一。在哺乳动物和扁虱中，A. 吞噬细胞分别存在于中性粒细胞和唾液腺中。尽管有大量研究表明 都专注于了解吞噬细胞菌在哺乳动物中生存所使用的策略。 细胞，相对较少的研究清楚地定义了吞噬细胞菌使用的分子策略。 在扁虱身上生存。在这个项目中，我们将对硬蜱进行全面的分子分析 肩胛骨有机阴离子转运多肽及其色氨酸相关基因 巨噬细胞与壁虱相互作用中的代谢途径。这项研究是建立在我们在 展示了吞噬弧菌如何调节基因表达和细胞信号转导以维持其在 向量。例如，我们以前的研究表明，吞噬细胞门氏菌调节扁虱的抗冻剂。 基因表达和肌动蛋白磷酸化是其在载体中存活的关键。在这里，我们提供强有力的 初步的体外和体内数据表明，吞噬弧菌可以诱导一种特定的 OATP(OATP4056)和犬尿氨酸转氨酶(KAT)，这是一个参与色氨酸途径的基因。 我们现在假设，OATP4056和KAT之间的相互作用不仅有助于A。 吞噬细胞在载体中存活，但也有助于将这种细菌传播到 脊椎动物宿主。RNAi分析表明，OATP4056基因的敲除对A. 吞噬细胞素的获取，但影响了它的存活和从这些扁虱传播。电泳法 用未感染和A。 感染吞噬细胞素的扁虱提供了进一步的证据，证明这种细菌的存在会影响 OATP4056在这些硬体中的表达。此外，我们发现有证据表明，色氨酸的一种代谢物 途径调节吞噬细胞在这些硬蜱中的存活和OATP4056基因的表达。这些 研究结果为我们提出的研究提供了重要的见解，以确定两者之间关系的分子基础 带扁虱的吞噬细胞群。基于我们强大的初步结果和提出的实验，我们 我相信这可能是一项变革性的研究，不仅可以作为研究亲密关系的模型 病原体与其节肢动物媒介建立的关系，但也可能导致发育 新的策略来阻断这种或其他医学立克次体物种的传播 重要性。