Effect of disease progression on the podocyte cell cycle in Alport Syndrome

Kidney Int. 2022 Jan;101(1):106-118. doi: 10.1016/j.kint.2021.08.026. Epub 2021 Sep 23.

Abstract

Progression of glomerulosclerosis is associated with loss of podocytes with subsequent glomerular tuft instability. It is thought that a diminished number of podocytes may be able to preserve tuft stability through cell hypertrophy associated with cell cycle reentry. At the same time, reentry into the cell cycle risks podocyte detachment if podocytes cross the G1/S checkpoint and undergo abortive cytokinesis. In order to study cell cycle dynamics during chronic kidney disease (CKD) development, we used a FUCCI model (fluorescence ubiquitination-based cell cycle indicator) of mice with X-linked Alport Syndrome. This model exhibits progressive CKD and expresses fluorescent reporters of cell cycle stage exclusively in podocytes. With the development of CKD, an increasing fraction of podocytes in vivo were found to be in G1 or later cell cycle stages. Podocytes in G1 and G2 were hypertrophic. Heterozygous female mice, with milder manifestations of CKD, showed G1 fraction numbers intermediate between wild-type and male Alport mice. Proteomic analysis of podocytes in different cell cycle phases showed differences in cytoskeleton reorganization and metabolic processes between G0 and G1 in disease. Additionally, in vitro experiments confirmed that damaged podocytes reentered the cell cycle comparable to podocytes in vivo. Importantly, we confirmed the upregulation of PDlim2, a highly expressed protein in podocytes in G1, in a patient with Alport Syndrome, confirming our proteomics data in the human setting. Thus, our data showed that in the Alport model of progressive CKD, podocyte cell cycle distribution is altered, suggesting that cell cycle manipulation approaches may have a role in the treatment of various progressive glomerular diseases characterized by podocytopenia.

Keywords: Alport syndrome; chronic kidney disease; podocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Cell Cycle
  • Disease Progression
  • Female
  • Humans
  • LIM Domain Proteins / metabolism
  • Male
  • Mice
  • Microfilament Proteins / metabolism
  • Nephritis, Hereditary* / genetics
  • Nephritis, Hereditary* / metabolism
  • Podocytes* / metabolism
  • Proteomics

Substances

  • Adaptor Proteins, Signal Transducing
  • LIM Domain Proteins
  • Microfilament Proteins
  • PDLIM2 protein, human
  • Pdlim2 protein, mouse