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podocyte cytoskeleton in the integrity of glomerular filtration is well established, the specific contribution of an upstream regulatory network comprising the Rho family of small GTPases\u2014modulators of the actin cytoskeleton\u2014is poorly characterized. The authors provide a comprehensive map of the podocyte Rho GTPase affinity interactome and identify a small GTPase-activating protein, SRGAP1, as a podocyte-specific RhoGAP. Through\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      models of experimental FSGS, they demonstrate that SRGAP1 prevents podocyte foot-process effacement. They also show that SRGAP1 mediates spatial restriction of the activity of the Rho GTPase RAC1, thereby maintaining morphologic plasticity in disease conditions. These findings indicate that a comprehensive understanding of the regulatory networks of small Rho GTPases is needed for precisely targeted therapeutic interventions in glomerular diseases.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Previous research demonstrated that small Rho GTPases, modulators of the actin cytoskeleton, are drivers of podocyte foot-process effacement in glomerular diseases, such as FSGS. However, a comprehensive understanding of the regulatory networks of small Rho GTPases in podocytes is lacking.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We conducted an analysis of podocyte transcriptome and proteome datasets for Rho GTPases; mapped\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      , podocyte-specific Rho GTPase affinity networks; and examined conditional knockout mice and murine disease models targeting\n                      <jats:italic toggle=\"yes\">Srgap1<\/jats:italic>\n                      . To evaluate podocyte foot-process morphology, we used super-resolution microscopy and electron microscopy;\n                      <jats:italic toggle=\"yes\">in situ<\/jats:italic>\n                      proximity ligation assays were used to determine the subcellular localization of the small GTPase-activating protein SRGAP1. We performed functional analysis of CRISPR\/Cas9-generated\n                      <jats:italic toggle=\"yes\">SRGAP1<\/jats:italic>\n                      knockout podocytes in two-dimensional and three-dimensional cultures and quantitative interaction proteomics.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      We demonstrated SRGAP1 localization to podocyte foot processes\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      and to cellular protrusions\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      .\n                      <jats:italic toggle=\"yes\">Srgap1fl\/fl*Six2Cre<\/jats:italic>\n                      but not\n                      <jats:italic toggle=\"yes\">Srgap1fl\/fl*hNPHS2Cre<\/jats:italic>\n                      knockout mice developed an FSGS-like phenotype at adulthood. Podocyte-specific deletion of\n                      <jats:italic toggle=\"yes\">Srgap1<\/jats:italic>\n                      by\n                      <jats:italic toggle=\"yes\">hNPHS2Cre<\/jats:italic>\n                      resulted in increased susceptibility to doxorubicin-induced nephropathy. Detailed analysis demonstrated significant effacement of podocyte foot processes. Furthermore,\n                      <jats:italic toggle=\"yes\">SRGAP1<\/jats:italic>\n                      -knockout podocytes showed excessive protrusion formation and disinhibition of the small Rho GTPase machinery\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      . Evaluation of a SRGAP1-dependent interactome revealed the involvement of SRGAP1 with protrusive and contractile actin networks. Analysis of glomerular biopsy specimens translated these findings toward human disease by displaying a pronounced redistribution of SRGAP1 in FSGS.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>SRGAP1, a podocyte-specific RhoGAP, controls podocyte foot-process architecture by limiting the activity of protrusive, branched actin networks. Therefore, elucidating the complex regulatory small Rho GTPase affinity network points to novel targets for potentially precise intervention in glomerular diseases.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1681\/asn.2020081126","type":"journal-article","created":{"date-parts":[[2021,1,29]],"date-time":"2021-01-29T14:00:15Z","timestamp":1611928815000},"page":"563-579","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":31,"title":["SRGAP1 Controls Small Rho GTPases To Regulate Podocyte Foot Process Maintenance"],"prefix":"10.1681","volume":"32","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6658-9159","authenticated-orcid":false,"given":"Manuel","family":"Rogg","sequence":"first","affiliation":[{"name":"Institute of Surgical Pathology, Faculty of Medicine, Medical Center \u2013 University of Freiburg, Freiburg, Germany"},{"name":"Department of Medicine IV, Faculty of Medicine, Medical Center \u2013 University of Freiburg, Freiburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jasmin I.","family":"Maier","sequence":"additional","affiliation":[{"name":"Institute of Surgical Pathology, Faculty of Medicine, Medical Center \u2013 University of Freiburg, Freiburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Dotzauer","sequence":"additional","affiliation":[{"name":"Department of Medicine IV, Faculty of Medicine, Medical Center \u2013 University of Freiburg, Freiburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadine","family":"Artelt","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, University Medicine Greifswald, Greifswald, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oliver","family":"Kretz","sequence":"additional","affiliation":[{"name":"III. 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