Project

Discipline

lange nicht-kodierende RNA; chronische Nierenerkrankung; Nierenfibrose

Lead
Die Nierenfibrose ist vergesellschaftet mit einer Reihe von verschiedenen Nierenerkrankungen. Seine Progression ist ein entscheidender Faktor für die Entwicklung der terminalen Niereninsuffizienz. Der zelluläre Ursprung von Matrix-produzierenden Zellen der Niere wurde in der Vergangenheit viel diskutiert. Kürzlich wurde in eleganten Fate-Tracing-Experimenten gezeigt, dass CD73+/platelet-derived growth factor receptor ß+ (PDGFR-ß) Perizyten die Hauptquelle für matrixproduzierende Zellen in der Niere sind. Die Mechanismen, die die Transdifferenzierung von Perizyten-zu-Myofibroblasten steuern, sind nicht vollständig aufgeklärt. Nicht-kodierende RNAs könnten wichtige Regulatoren dieses Prozesses sein. Lange nicht-kodierende RNAs (lncRNAs) sind neue wichtige epigenetische Regulatoren der zellulären Funktion.
Lay summary
Im vorliegenden Antrag isolierten und charakterisierten wir CD73 + / PDGFR-ß + -Zellen aus Mausnieren in einem Angiotensin-II-induzierten Nierenfibrose-Modell in Vorversuchen. Anschließend wurde isolierte RNA von CD73 + / PDGFR-ß + -Zellen einem globalen lncRNA-Expressionsprofil unterzogen. Die Antisense-lncRNA X zählte zu den am höchsten hochregulierten Kandidaten. Folgende spezifische Ziele werden im vorliegenden Antrag untersucht: 1. Analyse der lncRNA X Antisense-Transkript-Expression in Nierenbiopsien von Patienten mit verschiedenen Ätiologien der chronischen Nierenerkrankungen. 2. Entwicklung eines neuen lncRNA X  - basierten therapeutischen Ansatzes zur Verbesserung der Fibroseentwicklung in 2 etablierten Fibrosemodellen: Ang II - induzierte Fibrose und Nierenfibrose bei unilateraler Ureterobstruktion 3. Untersuchung der mechanistischen Rolle von lncRNA X in vitro sowie seiner transkriptionellen Regulatoren und Zielgene.

Direct link to Lay Summary

Last update: 09.04.2018

Natural persons

Self-organised

Title	Year

Renal fibrosis is commonly associated with various etiologies of chronic kidney disease. Its progression is a major determinant of end-stage renal disease. The cellular origin of matrix-producing cells of the kidney has been a matter of debate. Recently, it was shown in elegant fate-tracing experiments, that CD73+/platelet-derived growth factor receptor ß+ (PDGFR-ß) pericytes, are the major source of matrix-producing cells in the kidney. The mechanisms driving pericyte-to-myofibroblast transition are not fully elucidated. Non-coding RNAs might be important regulators of this process. Long non-coding RNAs (lncRNAs) are novel important epigenetic regulators of cellular function through various mechanisms, including the regulation of chromatin architecture, binding of protein and microRNA and directing of transcription factors to their sites of action. The overall aim of our proposal is to provide an in-depth analysis of lncRNA deregulation in isolated CD73+/PDGFR-ß+ cells as well as their function and therapeutic potential regarding fibrosis development in established murine models of kidney fibrosis and human kidney biopsies using state of the art molecular biology techniques. To this end we isolated and characterized CD73+/PDGFR-ß+-cells from mouse kidneys in an angiotensin II - induced kidney fibrosis model in preliminary experiments. Subsequently, isolated RNA of CD73+/PDGFR-ß+ cells was subjected to global lncRNA expression profiling. The antisense lncRNA X was among the most highly upregulated candidates. The following specific aims will be addressed:1.To analyse lncRNA X antisense transcript expression in renal biopsies of patients with various etiologies of chronic kidney disease to understand its regulation in human kidney diseases.2.To develop a novel lncRNA X (see below) - based therapeutic approach to ameliorate fibrosis development in 2 established fibrosis models: Ang II-induced fibrosis and renal fibrosis related to unilateral ureteral obstruction 3.To investigate the mechanistic role of lncRNA X in vitro and to determine its upstream transcriptional regulators and downstream target genes mediating pathological signalling. The role of the lncRNA X in promoting a phenotypic switch in pericyte-to- myofibroblast transdifferentiation, thereby resulting in fibrosis development, will be investigated. LncRNA X expresses several binding sites for microRNAs, thereby functioning as a microRNA sponge. Interestingly, our microRNA of interest is known to bioinformatically target several regulators of fibroblast activation. The lncRNA-microRNA interaction as well as regulation of target genes will be investigated in vitro and in vivo. In addition, lncRNA X shuttling into small vesicles will be assessed. Our analyses will provide important insights into the mechanisms driving pericyte-to-myofibroblast transition and may result in improved therapy of patients with chronic fibrotic kidney disease.