Acute kidney injury is a diagnosis associated with significant morbidity and mortality in the Veteran population. Therefore, we propose to develop a regenerative treatment for acute kidney injury based on progenitor cell exosomes. Recent studies suggest that the therapeutic effects of stem or progenitor cells are at least partially mediated through exosomes, a subset of extracellular vesicle. Exosomes are advantageous as they can be lyophilized for storage, are generally non-immunogenic, and do not require culture of cells just prior to therapeutic administration. The stem or progenitor cell type that will be utilized for this study is the induced nephron progenitor cell. Nephron progenitor cells form most of the differentiated cell types of the adult nephron, but are exhausted just prior to birth in humans. Induced nephron progenitors are derived from adult human cells in tissue culture through transcription factor reprogramming with inducible piggyBac transposons. The reprogramming factors are SNAI2, EYA1, and SIX1. We have isolated and characterized exosomes from induced nephron progenitor cells and find them to be abundant. Our preliminary data demonstrate that induced nephron progenitor exosomes have regenerative and protective properties in the HK-2 tubule cell line, human kidney organoids, and mouse models of acute kidney injury. However, further development is necessary to bring this potential therapeutic forward from bench to bedside. In Aim 1, we hypothesize that the exosomes will reduce markers of kidney injury in ischemia reperfusion and rhabdomyolysis mouse models of acute kidney injury. We will administer the induced nephron progenitor exosomes before, concurrent with, immediately after, and one day following injury and evaluate histology as well as markers of kidney function to determine the therapeutic effects on acute kidney injury. In Aim 2, we hypothesize that the induced nephron progenitor cell- derived exosomes protect human tubule cells from apoptosis induced by nephrotoxicants through specific protein and/or RNA species that can be modulated through genome engineering strategies to further enhance nephron progenitor extracellular vesicle efficacy. These studies will accelerate the translation of next-generation extracellular vesicle therapies based on advanced cell and genome engineering techniques for treatment of acute kidney injury.