Cre/loxP approach-mediated downregulation of Pik3c3 inhibits the hypertrophic growth of renal proximal tubule cells

J Cell Physiol. 2020 Dec;235(12):9958-9973. doi: 10.1002/jcp.29811. Epub 2020 May 31.


Nephron loss stimulates residual functioning nephrons to undergo compensatory growth. Excessive nephron growth may be a maladaptive response that sets the stage for progressive nephron damage, leading to kidney failure. To date, however, the mechanism of nephron growth remains incompletely understood. Our previous study revealed that class III phosphatidylinositol-3-kinase (Pik3c3) is activated in the remaining kidney after unilateral nephrectomy (UNX)-induced nephron loss, but previous studies failed to generate a Pik3c3 gene knockout animal model. Global Pik3c3 deletion results in embryonic lethality. Given that renal proximal tubule cells make up the bulk of the kidney and undergo the most prominent hypertrophic growth after UNX, in this study we used Cre-loxP-based approaches to demonstrate for the first time that tamoxifen-inducible SLC34a1 promoter-driven CreERT2 recombinase-mediated downregulation of Pik3c3 expression in renal proximal tubule cells alone is sufficient to inhibit UNX- or amino acid-induced hypertrophic nephron growth. Furthermore, our mechanistic studies unveiled that the SLC34a1-CreERT2 recombinase-mediated Pik3c3 downregulation inhibited UNX- or amino acid-stimulated lysosomal localization and signaling activation of mechanistic target of rapamycin complex 1 (mTORC1) in the renal proximal tubules. Moreover, our additional cell culture experiments using RNAi confirmed that knocking down Pik3c3 expression inhibited amino acid-stimulated mTORC1 signaling and blunted cellular growth in primary cultures of renal proximal tubule cells. Together, both our in vivo and in vitro experimental results indicate that Pik3c3 is a major mechanistic mediator responsible for sensing amino acid availability and initiating hypertrophic growth of renal proximal tubule cells by activation of the mTORC1-S6K1-rpS6 signaling pathway.

Keywords: class III phosphatidylinositol-3 kinase (Pik3c3); compensatory nephron hypertrophy (CNH); mammalian target of rapamycin complex 1 (mTORC1); renal proximal tubule cells (RPTC); ribosomal protein S6 (rpS6) phosphorylation; unilateral nephrectomy (UNX).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Class III Phosphatidylinositol 3-Kinases / antagonists & inhibitors
  • Class III Phosphatidylinositol 3-Kinases / genetics*
  • Extracellular Matrix Proteins / genetics
  • Gene Expression Regulation, Developmental / genetics
  • Humans
  • Integrases / genetics
  • Kidney / drug effects*
  • Kidney / growth & development
  • Kidney / pathology
  • Kidney / surgery
  • Kidney Tubules, Proximal / growth & development*
  • Kidney Tubules, Proximal / metabolism
  • Mechanistic Target of Rapamycin Complex 1 / genetics
  • Mice
  • Nephrectomy
  • Nephrons / growth & development*
  • Nephrons / metabolism
  • Phosphorylation / genetics
  • Protein-Lysine 6-Oxidase / genetics
  • Ribosomal Protein S6 Kinases, 90-kDa / genetics
  • Signal Transduction / drug effects
  • Sirolimus / pharmacology
  • Sodium-Phosphate Cotransporter Proteins, Type IIa / genetics*


  • Extracellular Matrix Proteins
  • Slc34a1 protein, mouse
  • Sodium-Phosphate Cotransporter Proteins, Type IIa
  • Lox protein, mouse
  • Protein-Lysine 6-Oxidase
  • Class III Phosphatidylinositol 3-Kinases
  • PIK3C3 protein, mouse
  • Mechanistic Target of Rapamycin Complex 1
  • Ribosomal Protein S6 Kinases, 90-kDa
  • Rps6ka1 protein, mouse
  • Cre recombinase
  • Integrases
  • Sirolimus