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.

项目摘要 疟疾引起的急性肾损伤（MAKI）是疟疾最严重的并发症之一， 成人和儿童死亡的最强指标。这种特殊并发症的影响已经 在很大程度上认识不足，其研究被忽视。缺乏合适的模型系统来阐明 在疟疾感染期间导致肾损害的机制。目前的知识差距正在阻碍有效的执行- 我们致力于开发准确的诊断和有效的治疗方法来治疗MAKI。因此，迫切需要 开发实验模型以研究MAKI的分子机制。这项研究的长期目标是 开发MAKI的新疗法，并确定MAKI的早期和准确的生物标志物，以更好地诊断和 治疗这种疾病。本申请的总体目标是（i）建立所需的科学框架 在体外和体内研究MAKI和（ii）确定疟疾寄生虫 引起急性肾损伤。中心假设是恶性疟原虫感染的红细胞（Pf-1）破裂， iRBC）释放直接促进急性肾小管坏死（ATN）的组分-MAKI的标志。的 提出研究的基本原理是，为MAKI基础研究奠定基础， 揭示促进疟疾肾损害的分子机制将有助于 更敏感的诊断和有效的治疗MAKI。中心假设将通过以下方式进行检验： 两个具体目标：1）开发MAKI的生理小鼠模型以鉴定导致MAKI的分子途径， ATN;和2）使用体外测定鉴定MAKI的分子机制。在第一个目标下，小鼠将不- 在感染疟原虫-啮齿类动物之前进行肾脏手术将通过以下方式评价肾功能： 测定尿量、肾小球滤过率、血尿素氮、血清肌酐水平和表达 急性肾损伤的生物标志物。将收获肾组织用于分析细胞死亡途径活化。 以及氧化应激水平。对于第二个目的，原代人肾小管和微血管 细胞将使用新技术（器官芯片）共同培养，以模仿孩子的自然结构， 尼。将培养物与Pf-iRBC破裂后释放的内容物一起孵育，以研究影响 对内皮-上皮屏障完整性、肾小管细胞毒性和细胞死亡途径的激活的影响。再- 这项申请中提出的研究是创新的，因为它将为科学界提供新的生理学， MAKI的校准模型。这些模型将不涉及使用高等脊椎动物，这使得它们能够获得- 适用于大多数实验室，与其他尝试的模型不同，这些模型是针对人肾 与疟疾有关的病理学。这项研究意义重大，因为它有望提供一个 强有力的科学基础，以促进对参与这种病理的信号通路的理解， 奥吉。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。