The generation and systematic standardized analysis of research models at an organismal level is fundamental for all projects within this consortium. This will be coordinated by the central project CP2. B. Zevnik, head of the in vivo Research Facility (ivRF) at CECAD and his team of the Transgene Service Unit, will support each project to develop sophisticated novel models to investigate podocyte biology and pathophysiology in vivo. Heighly advanced genetic approaches include the combination of distinct DNA recombinases, multicistronic knock-in strategies with 2a-peptides and CRISPR/Cas9 technologies as well as AAV gene transfer and, ultimately, aim to delete or precidely modify any gene of interest or to re-express transgenes in a time- and cost-efficient manner in podocytes exclusively. This in vivo pipeline will be instrumental to fulfill our mission to explore novel therapeutic approaches to treat FSGS. The phenotypic analysis of animal models requires state-of-the-art imaging encompassing electron microscopy as well as light microscopy of histological sections. J.U. Becker, a reknown nephropathologist at UoC, will evaluate all kidney sections offering a reference nephropathology for all animal models. We envision that organizing acquisition and analysis of images as a central core project will add a significant value to the single projects. This will be coordinated by M. Höhne together with A. Schauss.
In general, a clear data management strategy for data generation, data integration and data exchange will be essential to facilitate the communication and exchange between the individual projects. Thus, a central data platform will be provided by CP2 that covers all original data and information on (1) gene targeting, animal genotypes, phenotypes and treatment protocols, (2) histological data and reports similar to CP3, and (3) confocal/multiphoton/STED/EM imaging data.
Project related publications
Höhne, M., Ising, C., Hagmann, H., Völker, L., Brähler, S., Schermer, B., Brinkkötter, P.T., Benzing, T. Light Microscopic Visualization of Podocyte Ultrastructure Demonstrates Oscillating Glomerular Contractions. Am. J. Pathol. 2013 182, 332–338.
Koehler, S., Brähler, S., Braun, F., Hagmann, H., Rinschen, M.M., Späth, M.R., Höhne, M., Wunderlich, F.T., Schermer, B., Benzing, T., Brinkkoetter, P.T. Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes. Kidney International 2017 91, 1510–1517.
Agustian, P.A., Schiffer, M., Gwinner, W., Schäfer, I., Theophile, K., Modde, F., Bockmeyer, C.L., Traeder, J., Lehmann, U., Grosshennig, A., Kreipe, H.H., Bröcker, V., Becker, J.U. Diminished met signaling in podocytes contributes to the development of podocytopenia in transplant glomerulopathy. Am. J. Pathol. 178, 2007–2019 (2011).
Agustian, P.A., Bockmeyer, C.L., Modde, F., Wittig, J., Heinemann, F.M., Brundiers, S., Dämmrich, M.E., Schwarz, A., Birschmann, I., Suwelack, B., Jindra, P.T., Ahlenstiel, T., Wohlschläger, J., Vester, U., Ganzenmüller, T., Zilian, E., Feldkamp, T., Spieker, T., Immenschuh, S., Kreipe, H.H., Bröcker, V., Becker, J.U. Glomerular mRNA expression of prothrombotic and antithrombotic factors in renal transplants with thrombotic microangiopathy. Transplantation 95, 1242–1248 (2013).
Becker, J. U., Hoerning, A., Schmid, K. W. & Hoyer, P. F. Immigrating progenitor cells contribute to human podocyte turnover. Kidney Int. 72, 1468–1473 (2007).
Bockmeyer, C.L., Säuberlich, K., Wittig, J., Eßer, M., Roeder, S.S., Vester, U., Hoyer, P.F., Agustian, P.A., Zeuschner, P., Amann, K., Daniel, C., Becker, J.U. Comparison of different normalization strategies for the analysis of glomerular microRNAs in IgA nephropathy. Sci Rep 6, 31992 (2016).
Meyer-Schwesinger, C., Dehde, S., Klug, P., Becker, J.U., Mathey, S., Arefi, K., Balabanov, S., Venz, S., Endlich, K.H., Pekna, M., Gessner, J.E., Thaiss, F., Meyer, T.N. Nephrotic syndrome and subepithelial deposits in a mouse model of immune-mediated anti-podocyte glomerulonephritis. J. Immunol. 187, 3218–3229 (2011).
Meyer-Schwesinger, C., Lange, C., Bröcker, V., Agustian, P., Lehmann, U., Raabe, A., Brinkmeyer, M., Kobayashi, E., Schiffer, M., Büsche, G., Kreipe, H.H., Thaiss, F., Becker, J.U. Bone marrow-derived progenitor cells do not contribute to podocyte turnover in the puromycin aminoglycoside and renal ablation models in rats. Am. J. Pathol. 178, 494–499 (2011).
Modde, F., Agustian, P.A., Wittig, J., Dämmrich, M.E., Forstmeier, V., Vester, U., Ahlenstiel, T., Froede, K., Budde, U., Wingen, A.M., Schwarz, A., Lovric, S., Kielstein, J.T., Bergmann, C., Bachmann, N., Nagel, M., Kreipe, H.H., Bröcker, V., Bockmeyer, C.L., Becker, J.U. Comprehensive analysis of glomerular mRNA expression of pro- and antithrombotic genes in atypical haemolytic-uremic syndrome (aHUS). Virchows Arch. 462, 455–464 (2013).
Worthmann, K., Peters, I., Kümpers, P., Saleem, M., Becker, J.U., Agustian, P.A., Achenbach, J., Haller, H., Schiffer, M. Urinary excretion of IGFBP-1 and -3 correlates with disease activity and differentiates focal segmental glomerulosclerosis and minimal change disease. Growth Factors 28, 129–138 (2010).