O12 - Cell-Based Delivery of Nrf2-Loaded Extracellular Vesicles for Therapeutic Targeting of Acute Kidney Injury

Introduction: : Acute kidney injury (AKI) is a common and critical condition characterized by a rapid decline in renal function, often resulting from ischemia, toxins, or sepsis. AKI is associated with high morbidity, mortality, and increased risk of progression to chronic kidney disease, yet effective therapies remain limited. The lack of targeted treatments underscores the urgent need for novel therapeutic strategies that can mitigate kidney damage, promote repair, and improve patient outcomes. Innovative approaches to delivering protective and regenerative agents to injured kidneys are therefore of significant clinical relevance.
We developed a therapeutic approach using a cell-based delivery system to deploy engineered extracellular vesicles (eEVs) to potentially deliver Nrf2 to the kidney. Extracellular vesicles, including exosomes and microvesicles, play crucial roles in intercellular communication and tissue homeostasis. By engineering EVs to encapsulate and deliver therapeutic cargo, we aim to enhance the targeted delivery through the inflammation-mediated homing of the cells on the injured kidneys

Materials and

Methods: : Myeloid cells were transfected via non-viral electroporation with Nrf2 plasmid DNA. The resulting eEVs were characterized for size distribution and concentration using nanoparticle tracking analysis, with Nrf2 expression quantified by qPCR. For in vitro experiments, renal proximal tubule cells (RPTCs) treated with Nrf2 EVs were analyzed for Sox9 expression via qPCR. Three in vivo models (type 2 diabetes, rhabdomyolysis-induced AKI, cisplatin-induced AKI) received intravenous MemGlow-labeled MDSCs. Tissues (brain, lungs, heart, spleen, liver, kidneys), blood, and urine were collected. Biodistribution was assessed via IVIS and confocal imaging. Kidney analyses included histology, immunofluorescence, gene expression of injury markers (Kim-1, NGAL) and inflammatory markers (IL-6, IL-1β) via qPCR, and blood urea nitrogen (BUN) evaluation.

Results, Conclusions, and Discussions:: Myeloid cells expressed high levels of Nrf2 following non-viral electroporation and confocal imaging of fluorescently labeled eEVs showed efficient uptake by mouse proximal tubule cells. qRT-PCR results confirmed the ability of eEVs to package Nrf2 mRNA content. In vivo, when labeled myeloid cells were injected into db/db, rhabdomyolysis-induced, and cisplatin-induced AKI mice models, IVIS imaging revealed preferential homing and accumulation of these cells in the injured kidneys compared to non-injured kidneys, supporting the inflammation-mediated targeting strategy.
Our results demonstrate the feasibility of engineering immune cells that can effectively deliver Nrf2-loaded eEVs to AKI kidneys, targeting oxidative stress and inflammation. Our eEVs can overcome traditional challenges in EV therapy, including poor targeting by leveraging the inflammation-mediated homing capability of myeloid cells. These findings highlight the therapeutic potential of this approach for kidney injury. Future studies will further evaluate efficacy through histological analysis and additional functional metrics such as albumin-to-creatinine ratio (ACR) and creatinine levels, with the goal of translating this strategy to broader clinical applications in inflammation-driven conditions.

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