The Role of LIM-nebulette in podocyte mechanoresponse

PROJECT SUMMARY Podocyte abnormalities have been identified in many kidney diseases. The structural role of podocytes in the filtration barrier is known to be a dynamic process in which the podocyte actively restructures its actin cytoskeleton. Mechanical stress, such as severe hypertension, can have a detrimental impact on this process. LIM-nebulette is an actin binding protein within podocytes. We hypothesize that this protein helps podocytes adapt to the mechanical microenvironment in vivo. We have engineered an instrument with deformable surfaces to simulate the mechanical tensile forces experienced by podocytes in vivo. The goal of this project is to combine high-content image analysis with a novel mechanical stretch platform to understand how podocytes maintain their structural integrity against injury and use this system to study the operation of an important cell signaling pathway in the glomerular environment. Stretched and control samples will also be collected for fluorescent microscopy, protein expression, phosphproteomics and co-immunopreciptation to determine nebulette’s signaling network. Understanding the role of LIM-nebulette in maintaining podocyte cytoskeletal integrity could potentially reveal new avenues for stabilizing these cells and, consequently, preserving kidney function. By compiling and analyzing this data, we aim to clarify the role of LIM-nebulette in protecting podocytes. This project will yield cellular level insights into podocyte activity under glomerular environmental stress conditions by examining morphometric changes and alterations in mechanobiological signaling that may have an impact on chronic kidney disease.