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.

项目总结 疟疾所致急性肾损伤(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。这一特殊并发症的影响是 在很大程度上被忽视了，它的研究也被忽视了。目前还缺乏合适的模型系统来阐明 疟疾感染过程中导致肾脏损害的机制。目前的知识差距正在阻碍Ef-Ef Forts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 建立实验模型，研究MAKI的分子机制。这项研究的长期目标是 为MAKI开发新的治疗方法，并识别MAKI的早期和准确的生物标志物，以便更好地诊断和 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 体内外研究MAKI和(II)确定疟疾寄生虫致病的分子机制(S) 造成急性肾损伤。中心假说是恶性疟原虫感染的红细胞(Pf-1)破裂。 IRBC)释放的成分直接促进急性肾小管坏死(ATN)--MAKI的标志。这个 这项拟议研究的基本原理是，为Maki基础研究和 揭示促进疟疾肾脏损害的分子机制将促进 对Maki更敏感的诊断和有效的治疗。核心假说将通过追查来检验 两个具体目标：1)建立MAKI的生理性小鼠模型，以确定导致MAKI的分子通路 2)通过体外实验确定MAKI的分子机制。在第一个目标下，老鼠将不会- 在感染疟原虫-啮齿动物种之前进行德戈肾脏手术。肾功能将通过以下方式评估 测量尿量、肾小球滤过率、血尿素氮、血肌酐水平和表达 急性肾损伤的生物标志物。将采集肾组织用于分析细胞死亡途径的激活。 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(芯片上器官)进行共培养，以模仿孩子的自然结构- 奈伊。将培养物与PF-iRBC爆裂时释放的内容物孵育，以观察其影响(S) 对内皮-上皮屏障完整性、肾小管细胞毒性和激活细胞死亡通路的影响。再一次- 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。这些模型将不涉及使用高等脊椎动物，这使得它们可以- 与其他尝试的模型不同，这些模型是特定于人类肾脏的 与疟疾相关的病理学。这项拟议的研究具有重要意义，因为它有望提供一种 有坚实的科学基础来促进对参与此病的信号通路的了解。 奥奇。这项研究的结果将有助于开发创新的治疗方法和诊断测试 这将最终降低成人和儿童的疟疾死亡率。