Abstract: PO1273
ExoGAG, a New Method for Extracellular Vesicle and Glycoprotein Isolation in Urine That Unmasks the Pathophysiology of the Kidney and Identifies New Biomarkers of Kidney Disease
Session Information
- Cystic Kidney Disease - II
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1001 Genetic Diseases of the Kidneys: Cystic
Authors
- Vizoso, Marta, Nephrology Laboratory, Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain
- Lago, Nerea, Obesidomic Laboratory, Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain
- Camino, Tamara, Obesidomic Laboratory, Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain
- Lamas-Gonzalez, Olaya, Nephrology Laboratory, Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain
- Bravo, Susana, Proteomic Group, Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain
- Vazquez, Carmen, Nephrology Service, University Hospital of Santiago de Compostela, Santiago de Compostela, Spain
- Fidalgo diaz, Manuel, Nephrology Service, University Hospital of Santiago de Compostela, Santiago de Compostela, Spain
- Rodríguez, Cándido Díaz, Nephrology Service, University Hospital of Santiago de Compostela, Santiago de Compostela, Spain
- Pardo, Maria, Obesidomic Laboratory, Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain
- Garcia-Gonzalez, Miguel A., Nephrology Laboratory, Health Research Institute of Santiago de Compostela, Santiago de Compostela, Spain
Background
Glycosaminoglycans (GAGs) are large polysaccharides that interact through glycosidic bonds with proteins and lipids, forming the extracellular matrix; or with secreted proteins, as uromodulin. Glycosylation is altered in pathologies, as cancer or kidney diseases.
GAGs are present in extracellular vesicles (VEs), nanometric structures delimited by lipid bilayer that cells release and whose charge (RNA/miRNA, DNA and proteins) is essential in intercellular communication.
Our group developed a method for GAG, glycoproteins and VEs isolation in any biological sample, called ExoGAG (commercialized by Nasas Biotech), which led us to identify and characterize new signalling mechanisms, and identify new prognostic/diagnosis biomarkers, for example, in polycystic kidney disease (PKD).
Methods
Urine samples have been collected from patients genetically diagnosed with type I and II PKD at different stages of disease. Using ExoGAG, GAG-glycoprotein-VEs complex has been isolated and characterized by different proteomic techniques (Western Blot, mass spectrometry), gene expression (RT-PCR), and image characterization (electron microscopy, immunofluorescence).
Results
ExoGAG has allowed us to identify a new biomarkers in urine (in protection) in PKD patients, which are altered in disease progression, even anticipating currently used kidney damage markers. The characterization of these complexes has led us to discover signalling mechanisms between the different segments of the nephron, and whose function is altered in different pathologies. These findings have served other researchers deepen knowledge in different specialties such as Oncology and Endocrinology.
Conclusion
This new method for isolating the fraction associated with GAG in urine samples has allowed us to identify prognostic/diagnostic biomarkers of kidney diseases, based on glycoprotein and vesicular profile. Likewise, it has led us to identify new signalling mechanisms of the nephron, which opens a new field for a better understanding of renal pathophysiology. These results uncovered the potential as a method of EVs isolation for its use in the research of new cellular communication pathways or cellular mechanisms.