Direct reprogramming to human nephron progenitor-like cells using inducible piggyBac transposon expression of SNAI2-EYA1-SIX1

Kidney Int. 2019 May;95(5):1153-1166. doi: 10.1016/j.kint.2018.11.041. Epub 2019 Feb 28.

Abstract

All nephrons in the mammalian kidney arise from a transient nephron progenitor population that is lost close to the time of birth. The generation of new nephron progenitors and their maintenance in culture are central to the success of kidney regenerative strategies. Using a lentiviral screening approach, we previously generated a human induced nephron progenitor-like state in vitro using a pool of six transcription factors. Here, we sought to develop a more efficient approach for direct reprogramming of human cells that could be applied in vivo. PiggyBac transposons are a non-viral integrating gene delivery system that is suitable for in vivo use and allows for simultaneous delivery of multiple genes. Using an inducible piggyBac transposon system, we optimized a protocol for the direct reprogramming of HK2 cells to induced nephron progenitor-like cells with expression of only 3 transcription factors (SNAI2, EYA1, and SIX1). Culture in conditions supportive of the nephron progenitor state further increased the expression of nephron progenitor genes. The refined protocol was then applied to primary human renal epithelial cells, which integrated into developing nephron structures in vitro and in vivo. Such inducible reprogramming to nephron progenitor-like cells could facilitate direct cellular reprogramming for kidney regeneration.

Keywords: cap mesenchyme; direct reprogramming; nephron progenitor; reprogramming; transposon.

Publication types

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

MeSH terms

  • Cells, Cultured
  • Cellular Reprogramming / genetics*
  • DNA Transposable Elements / genetics*
  • Gene Transfer Techniques
  • Genetic Engineering / methods*
  • Homeodomain Proteins / genetics
  • Humans
  • Intracellular Signaling Peptides and Proteins / genetics
  • Nephrons / physiology*
  • Nuclear Proteins / genetics
  • Primary Cell Culture
  • Protein Tyrosine Phosphatases / genetics
  • Regeneration / genetics*
  • Snail Family Transcription Factors / genetics

Substances

  • DNA Transposable Elements
  • Homeodomain Proteins
  • Intracellular Signaling Peptides and Proteins
  • Nuclear Proteins
  • SIX1 protein, human
  • SNAI2 protein, human
  • Snail Family Transcription Factors
  • EYA1 protein, human
  • Protein Tyrosine Phosphatases