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                <title xml:lang="en">In vitro 2D and 3D kidney models to study the nephrotoxic response of uranium exposure</title>
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                <term xml:lang="en">Kidney models</term>
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              <p>As a heavy metal and alpha emitter, uranium presents chemical and radiological toxicity risks, accumulating preferentially in the kidneys, specifically in proximal convoluted tubules. The biological mechanisms and pathways impacted by uranium exposure can be formulated into an adverse outcome pathway (AOP) (https://aopwiki.org/aops/447), allowing us to identify the different steps and gaps between an initiating event to the kidney toxicity. Using in vitro models of renal proximal tubule epithelial cells (hRPTEC TERT1) or induced kidney organoids (iKO), this study aims to reinforce and contribute to the development of the AOP of kidney toxicity. After identifying the U(VI) concentrations that induce adverse outcomes (apoptosis or necrosis), key events linked to oxidative stress, apoptosis, survival, inflammation, and renal damage are studied at the gene and protein levels. Apoptosis (Casp 3/7, Bax/Bcl2, cytochrome C) is significantly induced starting from exposure to 300 µM for hRPTEC and from 500µM for iKO, and necrosis (LDH) from 500 µM for both models. A time-dependent rise in ROS production and an antioxidant response -increased HO-1, NQO-1, GCLC- are observed in hRPTEC cells. Whereas SOD2, NQO-1 are augmented in iKO but GCLC, CYP2E1 and CAT are diminished in iKO. Uranium concentrations &amp;gt;300µM induced a marked inflammatory response in both models, with increased levels of TNFα, IL-6 and IL-18 in hRPTEC cells and only TNFα in iKO.  The study of renal damage markers revealed a slight increase in KIM-1 at 500 µM and a decreased of Collagen IV in both models and an increased level of Calbindin in iKO. In conclusion, this work provides an insight into the key events in the AOP of uranium-induced renal failure featuring a human phenotype of proximal tubule epithelial cells and kidney organoid model, while considering the dose-dependent effect.</p>
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