[R01] Alternative splicing of UMOD as a protective mechanism for thick ascending cells in kidney injury.

Ente: National Institute of Diabetes and Digestive and Kidney Diseases

Scadenza: 2030-03-31

Importo max: 674.691 EUR

Paese: US

Descrizione

Acute kidney injury (AKI) is a major determinant of mortality and morbidity in hospitalized patients. Unfortunately, AKI remains without specific therapy. Uromodulin (UMOD, also known as Tamm-Horsfall protein, gene name UMOD) is a kidney-specific protein uniquely expressed in the cells of thick ascending limbs (TAL). Uromodulin is secreted in the urine and circulation and is implicated in modulating kidney injury, but the functions of intracellular uromodulin remain poorly understood. We discovered a splice variant of UMOD (AS- UMOD) that is directed intracellularly within TAL cells. Our preliminary data suggest that AS-UMOD is induced by injury and is targeted towards the inner mitochondrial membrane to protect TAL cells by enhancing mitochondrial function. The goal of this application is to establish the importance of AS-UMOD induction as a protective mechanism within TAL cells against AKI. We aim to elucidate innovative concepts related to UMOD biology that have broad implications, enhance AS-UMOD expression using splice switching oligonucleotides (SSOs) as a potential novel therapeutic strategy for AKI and expand the understanding of AKI pathophysiology and the key role of AS-UMOD in regulating TAL metabolism. Our overarching hypothesis is that the splice variant AS-UMOD is induced by injury and protects TAL cells through intracellular targeting that enhances mitochondrial metabolism. We will investigate this hypothesis through the following aims Aim 1 will establish the spatiotemporal dynamics of AS-UMOD during AKI and its unique targeting to the mitochondria Aim 2 will demonstrate that AS-UMOD ameliorates mitochondrial function by enhancing the activity of SLC25 carriers across the inner membrane. Aim 2 will establish the protective functions and metabolic impact of AS-UMOD during AKI. This research will involve the use of innovative tools developed for these studies such as SSOs, customized real-time PCR primers and antibodies to detect AS-UMOD, innovative multiplexed fluorescence imaging and spatial metabolomics to define the spatial distribution across time and the metabolic impact of AS-UMOD. Various animal models of AKI (renal ischemia-reperfusion injury achieved through renal pedicle clamping, cisplatin toxicity or lipopolysaccharide injections) will be used. Relevant cell culture models will also be used to dissect mechanistic pathways that define the role of AS-UMOD. This research is innovative, because it will establish a novel protective role for this splice variant that is targeted intracellularly. Findings from this research may also have broad impact on cell biology and the role of splicing on cellular trafficking and cytoprotection. Our long-term goal is to use our increasing knowledge and expertise in uromodulin biology to develop therapeutic tools against AKI that could mitigate the severity of kidney injury and promote repair. Istituzione: INDIANA UNIVERSITY INDIANAPOLIS PI: Tarek Maurice Ashkar Progetto: 1R01DK146350-01A1

Settori: National Institute of Diabetes and Digestive and Kidney Diseases

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