New strategies to study malaria-induced acute kidney injury

研究疟疾引起的急性肾损伤的新策略

• 批准号: 10333643
• 负责人: Julio Gallego-Delgado
• 金额: $ 15.45万
• 依托单位: HERBERT H. LEHMAN COLLEGE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10333643
• 关键词: Acute Kidney Tubular Necrosis; Acute Renal Failure with Renal Papillary Necrosis; Adult; Apoptosis; Apoptotic; Architecture; Award; Basic Science; Biological Markers; Biological Models; Blood; Blood Urea Nitrogen; CASP3 gene; Case Fatality Rates; Cell Death; Cells; Cessation of life; Child; Cleaved cell; Communities; Complication; Creatinine; Data; Development; Diagnosis; Diagnostic tests; Disease; Early Diagnosis; Endothelium; Epithelial; Erythrocytes; Experimental Models; Foundations; Generations; Glomerular Filtration Rate; Glomerulonephritis; Goals; Harvest; Health; Heme; Hemoglobin; Home; Human; Immune; Immunoblotting; Immunofluorescence Immunologic; Impairment; In Vitro; Incubated; Infection; Injury; Injury to Kidney; Innovative Therapy; Interstitial Nephritis; Invaded; Kidney; Kinetics; Knowledge; Laboratories; Lead; Life Cycle Stages; Malaria; Measures; Mediating; Metabolic; Mission; Modeling; Molecular; Mus; Necrosis; Nephrectomy; Oliguria; Operative Surgical Procedures; Output; Oxidative Stress; Parasites; Pathology; Pathway interactions; Patients; Physiological; Pilot Projects; Plasmodium; Plasmodium berghei; Plasmodium falciparum; Proliferating; Proteome; Public Health; Publishing; Renal Tissue; Renal function; Reporting; Research; Research Project Grants; Rupture; Serum; Signal Pathway; Testing; Tissues; Tubular formation; United States National Institutes of Health; Uric Acid; Urine; Vertebrates; Wild Type Mouse; accurate diagnosis; base; cytotoxic; cytotoxicity; effective therapy; hemodynamics; improved; in vitro Assay; in vitro Model; in vivo; in vivo Model; innovation; kidney cell; malaria infection; mortality; mouse model; neglect; new technology; novel; novel therapeutics; organ on a chip; pediatric patients; physiologic model; tool

PROJECT SUMMARY Malaria-induced acute kidney injury (MAKI) is one of the most severe complications of malaria and the strongest indicator of death in both adults and children. The impact of this specific complication has been largely under-recognized, and its study neglected. There is a lack of suitable model systems for elucidating the mechanisms that lead to renal impairment during malaria infection. The current gap in knowledge is impeding ef- forts to develop accurate diagnoses and effective therapies to treat MAKI. There is, therefore, a critical need to develop experimental models to study the molecular mechanisms of MAKI. The long-term goal of this study is to develop new therapies for MAKI and identify early and accurate biomarkers of MAKI, to better diagnose and treat this disease. The overall objectives in this application are to (i) establish the scientific framework needed to study MAKI in vitro and in vivo and (ii) determine the molecular mechanism(s) by which malaria parasites induce acute kidney injury. The central hypothesis is that rupture of P. falciparum-infected red blood cells (Pf- iRBCs) releases components that directly promote acute tubular necrosis (ATN)—the hallmark of MAKI. The rationale that underlies the proposed research is that establishing the foundation for MAKI basic research and unveiling the molecular mechanisms that promote renal impairment in malaria will facilitate progress towards more sensitive diagnoses and effective treatments for MAKI. The central hypothesis will be tested by pursuing two specific aims: 1) Develop a physiological mouse model of MAKI to identify the molecular pathways leading to ATN; and 2) Identify molecular mechanisms of MAKI using in vitro assays. Under the first aim, mice will un- dergo renal surgery before infection with Plasmodium-rodent spp. The renal function will be evaluated by measuring urine output, glomerular rate filtration, blood urea nitrogen, serum creatinine levels and expression of biomarkers of acute kidney injury. Kidney tissues will be harvested for analysis of cell death pathway activa- tion as well as levels of oxidative stress. For the second aim, primary human renal tubular and microvascular cells will be co-cultured using novel technology (Organ-on-a-chip) to mimic the natural architecture of the kid- ney. Cultures will be incubated with the contents released upon bursting of Pf-iRBCs to investigate the effect(s) on endothelial-epithelial barrier integrity, tubular cytotoxicity and activation of cell death pathways. The re- search proposed in this application is innovative, as it will provide the scientific community with new physiologi- cal models of MAKI. The models will not involve the use of high vertebrate animals, which makes them acces- sible for most laboratories, and unlike other attempted models, these models are specific for the human renal pathology associated with malaria. The proposed research is significant because it is expected to provide a strong scientific basis on which to advance the understanding of the signaling pathways involved in this pathol- ogy. The results of this study will contribute to the development of innovative therapies and diagnostic tests for MAKI, which will ultimately reduce malaria mortality in both adults and children.

项目概要 疟疾引起的急性肾损伤（MAKI）是疟疾最严重的并发症之一，也是疟疾最严重的并发症之一。 这种特定并发症的影响是成人和儿童死亡的最强指标。 很大程度上没有得到充分认识，其研究也被忽视。缺乏合适的模型系统来阐明这一点。 疟疾感染期间导致肾功能损害的机制 目前的知识差距阻碍了研究的进展。 因此，迫切需要开发准确的诊断和有效的治疗方法来治疗 MAKI。 开发实验模型来研究MAKI的分子机制这项研究的长期目标是。 开发 MAKI 的新疗法并识别 MAKI 的早期和准确的生物标志物，以更好地诊断和治疗 本申请的总体目标是（i）建立所需的科学框架。 体外和体内研究 MAKI 并 (ii) 确定疟疾寄生虫的分子机制 诱发急性肾损伤。核心假设是恶性疟原虫感染的红细胞（Pf-）破裂。 iRBC）释放直接促进急性肾小管坏死（ATN）的成分——这是 MAKI 的标志。 拟议研究的基本原理是为 MAKI 基础研究和 揭示促进疟疾肾损伤的分子机制将有助于在这方面取得进展 对 MAKI 更敏感的诊断和有效的治疗将通过追求来检验中心假设。 两个具体目标：1) 开发 MAKI 生理小鼠模型，以确定分子主导途径 ATN；和 2) 使用体外试验确定 MAKI 的分子机制。 在感染疟原虫啮齿类动物之前进行肾手术将通过以下方式评估肾功能。 测量尿量、肾小球滤过率、血尿素氮、血清肌酐水平和表达 将收集急性肾损伤的生物标志物用于分析细胞死亡途径激活。 和氧化应激水平对于第二个目标，原代人肾小管和微血管。 细胞将使用新技术（器官芯片）进行共培养，以模仿儿童的自然结构 ney. 将用 Pf-iRBC 破裂时释放的内容物孵育培养物以研究效果。 内皮-上皮屏障完整性、肾小管细胞毒性和细胞死亡途径激活的影响。 本申请中提出的搜索是创新的，因为它将为科学界提供新的生理学 MAKI 的 cal 模型不会涉及使用高等脊椎动物，这使得它们能够 适用于大多数实验室，并且与尝试的其他模型不同，这些模型针对人类肾脏 与疟疾相关的病理学研究具有重要意义，因为它有望提供一种方法。 坚实的科学基础可以促进对这种病理所涉及的信号通路的理解 这项研究的结果将有助于开发创新疗法和诊断测试。 MAKI，最终将降低成人和儿童的疟疾死亡率。