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 st...