This project employs morphometric methods to evaluate kidney biopsy samples obtained under Project # Z01 DK 69062. One hundred twenty diabetic Pima Indians who participated in a randomized double-blinded, placebo-controlled clinical trial to test the renoprotective efficacy of the angiotensin receptor blocker losartan underwent percutaneous kidney biopsy. Tissue specimens from these biopsies are processed at the Beckman Center for Electron Microscopy at Stanford University. Digitized images of individual glomeruli are compiled to create photomontages of individual glomeruli and the montage is calibrated to give precise measurements of glomerular structure. The cell nuclei identified in the montage are numbered and labeled according to their cell type (endothelial, mesangial or podocyte) and measurements are performed to determine the average number of podocytes per glomerulus, using the method of Weibel. The volume fraction of podocytes within the glomerulus is determined by multiplying the average glomerular volume by the numerical volume density of the podocytes. Total filtration surface area is computed by estimating the total number of functioning glomeruli and multiplying it by the average filtration surface for each glomerulus measured. Hydraulic permeability of the glomerular capillary is calculated from measurements of filtration slit frequency and basement membrane thickness using the ultrastructural model of Drumond and Dean, and the single nephron ultrafiltration coefficient is computed. Previous work by our group suggests that podocyte loss is the principal structural change responsible for functional progression, characterized by increasing albuminuria and declining glomerular filtration rate.? In the past year, we completed morphometric measurements from the first 24 biopsies. In the coming year, we will continue making morphometric measurements on additional specimens. In addition, data from morphometric studies will be combined with data from physiologic studies obtained on the same research subjects to determine how morphologic changes in the glomeruli correlate with the progression of diabetic kidney disease and how blockers of angiotensin attenuate those changes. These data will also be combined with data generated from gene expression studies performed on the same tissue specimens to better understand how cellular processes lead to structural and functional changes.? The combination of structural, functional, and genetic studies may yield new insights into how diabetic nephropathy progresses, demonstrate how angiotensin blockers slow progression, and suggest novel therapies for diabetic kidney disease.
