Targeting Neph1 and ZO-1 protein-protein interaction in podocytes prevents podocyte injury and preserves glomerular filtration function

Sci Rep. 2017 Sep 21;7(1):12047. doi: 10.1038/s41598-017-12134-8.

Abstract

Targeting protein-protein interaction (PPI) is rapidly becoming an attractive alternative for drug development. While drug development commonly involves inhibiting a PPI, in this study, we show that stabilizing PPI may also be therapeutically beneficial. Junctional proteins Neph1 and ZO-1 and their interaction is an important determinant of the structural integrity of slit diaphragm, which is a critical component of kidney's filtration system. Since injury induces loss of this interaction, we hypothesized that strengthening this interaction may protect kidney's filtration barrier and preserve kidney function. In this study, Neph1-ZO-1 structural complex was screened for the presence of small druggable pockets formed from contributions from both proteins. One such pocket was identified and screened using a small molecule library. Isodesmosine (ISD) a rare naturally occurring amino acid and a biomarker for pulmonary arterial hypertension was selected as the best candidate and to establish the proof of concept, its ability to enhance Neph1-CD and ZO-1 binding was tested. Results from biochemical binding analysis showed that ISD enhanced Neph1 and ZO-1 interaction under in vitro and in vivo conditions. Importantly, ISD treated podocytes were resistant to injury-induced loss of transepithelial permeability. Finally, mouse and zebrafish studies show that ISD protects from injury-induced renal damage.

Publication types

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

MeSH terms

  • Animals
  • Cells, Cultured
  • Glomerular Filtration Rate / drug effects
  • Humans
  • Isodesmosine / pharmacology*
  • Kidney / drug effects
  • Kidney / metabolism
  • Kidney / physiopathology
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mice
  • Molecular Docking Simulation
  • Podocytes / drug effects*
  • Podocytes / metabolism
  • Protein Binding / drug effects
  • Protein Domains
  • Zebrafish
  • Zonula Occludens-1 Protein / chemistry
  • Zonula Occludens-1 Protein / genetics
  • Zonula Occludens-1 Protein / metabolism*

Substances

  • KIRREL1 protein, human
  • Membrane Proteins
  • TJP1 protein, human
  • Zonula Occludens-1 Protein
  • Isodesmosine